i 

United States 

Department of 

Agriculture

The State University of New York College of 

Environmental Science and Forestry Petition (19-309-

01p) for Determination of Nonregulated Status for 

Blight-Tolerant Darling 58 American Chestnut

(Castanea dentata) 

OECD Unique Identifier: 

ESF-DAR58-3

Draft Environmental Impact Statement 

July 2022 

Agency Contact 

Cindy Eck

USDA, APHIS

Biotechnology Regulatory Services 

4700 River Road

Riverdale, MD 20737 

The U.S. Department of Agriculture (USDA) prohibits discrimination in all its programs and activities on the basis of 

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opportunity provider and employer. 

Mention of companies or commercial products in this report does not imply recommendation or endorsement by the 

U.S. Department of Agriculture over others not mentioned. USDA neither guarantees nor warrants the standard of any 

product mentioned. Product names are mentioned solely to report factually on available data and to provide specific 

information. 

This publication reports research involving pesticides. All uses of pesticides must be registered by appropriate State 

and/or Federal agencies before they can be recommended. 
i

Executive Summary

The U.S. Department of Agriculture (USDA), Animal and Plant Health Inspection Service 

(APHIS) received a request (APHIS Petition 19-309-01p) from the State University of New 

York College of Environmental Science and Forestry (ESF) seeking a determination of 

nonregulated status for Darling 58 American chestnut (Castanea dentata (Marsh.) that has been 

engineered to be tolerant to the fungal pathogen, Cryphonectria parasitica (ESF 2019). When 

APHIS receives a petition for nonregulated status of an organism currently regulated under its 

PPA authority codified in 7 CFR part 340, the Agency is required to make a decision. As a 

Federal agency, APHIS must also comply with applicable U.S. environmental laws and 

regulations because a decision on a petition for nonregulated status, whether positive or negative, 

is a final Agency action that might cause environmental impacts.

The petition stated that APHIS should not regulate Darling 58 American chestnut because it does 

not present a plant pest risk. In the event of a determination of nonregulated status, the 

nonregulated status would include Darling 58 American chestnut and any progeny derived from 

crosses between Darling 58 American chestnut and sexually compatible species including 

American chestnut (C. dentata), Chinese chestnut (C. mollissima), Japanese chestnut (C. 

crenata), European chestnut (C. sativa), Chinese chinquapin chestnut (C. henryi), Allegheny 

Chinquapin chestnut (C. pumila), and Ozark Chinquapin Chestnut (C. ozarkensis) (ESF 2019).

Regulatory Authority 

The Plant Protection Act of 2000 (PPA), as amended (7 U.S.C. §§ 7701-7772), provides the legal 

authorization for the APHIS plant protection mission. It authorizes the Agency to regulate the 

introduction of potential plant pests into the territorial boundaries of the United States, and their 

interstate movement within U.S. boundaries by establishing quarantine, eradication and control 

programs. Implementing rules, regulations and guidelines for this enabling legislation (PPA) are 

codified in Title 7 of the U.S. Code of Federal Regulations (CFR). Rules that implement this 

authority specific to organisms developed using genetic engineering have been published in 7 

CFR part 340.

The regulations also provide a process to petition APHIS to determine that an organism

developed using genetic engineering is nonregulated. An organism is no longer subject to the 

requirements of 7 CFR part 340 if APHIS determines that it is unlikely to pose a plant pest risk. 

A determination of nonregulated status means that the regulated organism is no longer subject to 

the regulations in 7 CFR part 340 and, therefore, there is no longer any authority for APHIS to 

require a permit or notification for the importation, interstate movement, or environmental 

release of the organism pursuant to 7 CFR part 340.

Two other agencies, the Federal Drug Administration (FDA) and the Environmental Protection 

Agency (EPA), are involved in regulating organisms developed using genetic engineering. The 

regulatory roles of USDA-APHIS, the FDA, and the EPA are described by the “Coordinated 

Framework,” a 1986 policy statement from the Office of Science and Technology Policy that 

describes the comprehensive Federal policy for ensuring the safety of biotechnology research 

and products. 
ii

The FDA regulates organisms developed using genetic engineering under the authority of the 

Federal Food, Drug, and Cosmetic Act (FFDCA) (21 U.S.C. 301 et seq.). The FDA implements a 

voluntary consultation process to ensure that human food and animal feed safety issues or other 

regulatory issues, such as labeling, are resolved before commercial distribution of food derived 

from products developed using genetic engineering.

The EPA is responsible for regulating the sale, distribution, and use of pesticides, including 

pesticides that are produced by an organism through techniques of biotechnology. The EPA 

regulates plant incorporated protectants (PIPs) and microorganisms used as pesticides, e.g. 

bacteria, fungi, viruses, bacteriophages; both naturally occurring and genetically engineered

under the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) (7 U.S.C. 136 et seq.) 

and certain microorganisms under the Toxic Substances Control Act (15 U.S.C. 53 et seq.). 

Under FIFRA (7 U.S.C. 136 et seq.), the EPA regulates the use of pesticides (requiring 

registration of a pesticide for a specific use prior to distribution or sale of the pesticide for a 

proposed use pattern). The EPA examines the ingredients of the pesticide; the particular site or 

crop on which it is to be used; the amount, frequency, and timing of its use; and storage and 

disposal practices. Prior to registration of a new pesticide or a new use for a previously registered 

pesticide, the EPA must determine that the pesticide will not cause unreasonable adverse effects 

on the environment and a reasonable certainty of no harm to humans when used in accordance 

with label instructions. The EPA reevaluates all pesticides every fifteen years (or shorter) to 

ensure they meet current standards for continued safe use (7 U.S.C. 136a(g)(1)(A)(iv)). 

The EPA also sets tolerances for residues of pesticides on and in food and animal feed, or 

establishes an exemption from the requirement for a tolerance, under the FFDCA. The EPA is 

required, before establishing pesticide tolerance, to reach a safety determination based on a 

finding of reasonable certainty of no harm under the FFDCA, as amended by the Food Quality 

Protection Act (FQPA). The FDA enforces the pesticide tolerances set by the EPA.

Purpose and Need for Agency Action 

Under the authority of the plant pest provisions of the PPA and 7 CFR part 340, APHIS regulates

the safe development and use of organisms developed using genetic engineering. Any party can 

petition APHIS to seek a determination of nonregulated status for an organism developed using 

genetic engineering that is regulated under 7 CFR part 340. As required by 7 CFR part 340,

APHIS must respond to petitioners that request a determination of the regulated status of plants

or organisms developed using genetic engineering such as Darling 58 American chestnut. When 

a petition for nonregulated status is submitted, APHIS must make a determination if the 

organism is unlikely to pose a plant pest risk. The petitioner is required to provide information 

under 7 CFR part 340 related to plant pest risk that the agency may use to compare the plant pest 

risk of the regulated organism to that of the unmodified organism. An organism developed using 

genetic engineering is no longer subject to the regulatory requirements of 7 CFR part 340 when 

APHIS determines that it is unlikely to pose a plant pest risk. In November 2019, APHIS 

received a petition from ESF requesting a determination of the regulated status for Darling 58 

American chestnut. The purpose of the petition was to request nonregulated status for Darling 58 

American chestnut because it does not pose a plant pest risk. APHIS prepared a PPRA and this 
iii

EIS to respond to the request and avoid the inappropriate use of public resources regulating a 

product developed using genetic engineering if the agency determines that it has no authority to 

do so.

Consistent with the Council of Environmental Quality’s National Environmental Policy Act 

(NEPA) regulations and the USDA and APHIS NEPA implementing regulations and procedures

(40 CFR parts 1500-1508, 7 CFR part 1b, and 7 CFR part 372), APHIS has prepared this draft 

Environmental Impact Statement (EIS) to consider the potential environmental impacts of an 

agency determination of nonregulated status. Specifically, this draft EIS has been prepared in 

order to evaluate the impacts on the quality of the human environment1 that may result from a 

determination of nonregulated status of Darling 58 American chestnut.

Public Involvement 

APHIS sought comments for the petition that is the subject of this EIS in a Federal Register 

notice dated August 19, 2020. The docket received a total of 4,320 public comments. Of the 

4,320 comments received 1,619 comments were in opposition to deregulation of Darling 58 

American chestnut, 2,698 comments were in favor of deregulation, and 3 were out of scope. The 

issues that were raised in the opposing comments were in regard to environmental impacts of the 

unconfined release of a forest tree developed using genetic engineering, impacts to native 

communities, human health and safety impacts of using a wheat gene, the need for long term 

studies, the potential for chestnut to be more susceptible to chestnut blight as well as other 

diseases, the potential for impacts to organic producers, impacts to trade, and general antibiotech sentiments. The in favor of the petition comments emphasized the positive 

environmental and socio-economic benefits of restoring American chestnut to its original range.

As part of its scoping process to identify issues to address in this EIS, APHIS also published a 

Notice of Intent (NOI) to prepare an EIS and sought public input during a 30-day comment 

period (August 6 to September 7, 2021). Comments were submitted by individuals, academic 

researchers, and non-government organizations. The majority of comments submitted were 

opposed to a determination of nonregulated status for darling 58 American chestnut. 

Alternatives Analyzed 

In this EIS, APHIS considered two alternatives for its response to the ESF petition for 

nonregulated status. The two alternatives are: 1) continue to regulate Darling 58 American 

chestnut (No Action Alternative) and 2) approve the petition for nonregulated status of Darling 

58 American chestnut (Preferred Alternative). These alternatives are further described here and 

in Chapter 2.

Alternative 1: No Action Alternative—Continue as Regulated 

Under the No Action Alternative, APHIS would deny the petition because it was found to pose a 

plant pest risk. Darling 58 American chestnut and progeny derived from Darling 58 American 

1 Under NEPA regulations, the “human environment” includes “the natural and physical environment and the 

relationship of people with that environment” (40 CFR §1508.14).
iv

chestnut would continue to be regulated under the regulations at 7 CFR part 340. Any 

introduction of Darling 58 American chestnut would still require authorization by APHIS. In 

addition, measures to ensure physical and reproductive confinement of Darling 58 American 

chestnut would continue to be implemented for any existing or new authorization. APHIS might 

choose this alternative if there were insufficient evidence to demonstrate the lack of plant pest 

risk from the unconfined cultivation of Darling 58 American chestnut. 

This alternative is not the Preferred Alternative because APHIS has concluded through a PPRA

that Darling 58 American chestnut is unlikely to pose a plant pest risk (USDA-APHIS 2020). 

Choosing this alternative would not satisfy the purpose and need of making a determination of

plant pest risk status and responding to the petition for nonregulated status.

Alternative 2: Preferred Alternative – Determination that Darling 58 American chestnut is 

No Longer Regulated 

Under this alternative, Darling 58 American chestnut and progeny derived from it would no 

longer be regulated under the regulations at 7 CFR part 340. APHIS would no longer require 

authorizations for introductions of Darling 58 American chestnut and progeny derived from this 

event. This alternative best meets the purpose and need to respond appropriately to a petition for

nonregulated status based on the requirements in 7 CFR part 340 and the agency’s authority 

under the plant pest provisions of the PPA. Because the agency has concluded that Darling 58 

American chestnut is unlikely to pose a plant pest risk (USDA-APHIS 2020), a determination of 

nonregulated status of Darling 58 American chestnut is a response that is consistent with the 

plant pest provisions of the PPA, the regulations codified in 7 CFR part 340, and the 

biotechnology regulatory policies in the Coordinated Framework. A determination of 

nonregulated status and this EIS would not necessarily apply to other blight tolerant American 

chestnut events as APHIS’ regulatory practice is to review requests on a case-by-case basis.

Affected Environment and Environmental Consequences 

Although the Preferred Alternative would allow for new plantings of Darling 58 American 

chestnut to occur anywhere in the United States the petitioner has stated their intention is 

ecological restoration and that initial distribution will consist of long-term research plots and 

relatively small-scale public horticultural plantings and will focus on areas where there are 

surviving small remnant American chestnut populations. For this reason, APHIS considered the 

affected environment for this EIS to be those areas of the United States where American chestnut 

was once a dominant forest tree.

Environmental issues are assessed individually in Chapter 4. The scope of this EIS analyzes the 

potential for direct and indirect impacts that might result from a determination of nonregulated 

status of Darling 58 American chestnut.

APHIS determined that the potential planting of Darling 58 American chestnut for restoration 

purposes or for any other purpose is likely to have some degree of impact on the environment. If 

blight-tolerant American chestnut were to become established as an important canopy tree, it 

would begin to influence ecosystem structure and function in these areas, as it did prior to the 
v

blight. Impacts to air quality, surface water and groundwater, soils, biodiversity, and habitats are 

all possible from planting Darling 58 American chestnut. The degree of environmental impacts 

will depend on a variety of factors that include the geographic locale, local biota, weather, 

inherent soil characteristics, and prevalence and diversity of wildlife. Given the historically slow 

spread rates, low propagule pressure, and need for disturbance to provide sufficient light for fast 

growth, the rate of increase would likely be very slow. Without aggressive restoration efforts, 

requiring considerable effort and coordination at landscape scales, it may require centuries 

before American chestnut becomes a significant presence in the landscape (Gustafson et al. 

2017; Gustafson et al. 2018). 

While it is difficult to predict what impacts Darling 58 American chestnut will have on forest 

biodiversity, in the long term if American chestnut were to become a dominant species again, it 

is reasonable to believe there are likely to be positive impacts on the biodiversity of animal 

species as chestnut provides a more stable and more abundant source of mast than oaks, hickory, 

and beech species (Diamond et al. 2000) while decreasing the abundance of some tree species 

such as oaks. As American chestnut grows faster than other hardwood species, there may be 

positive impacts on climate change through greater carbon sequestration. Species that became 

more prevalent during the demise of chestnut, such as oak and maple, may become less 

prevalent.

Chestnuts are the most abundant treenut after coconut (Davison et al. 2021a). The industry is 

centered in China (85% of world production) and is largely absent in the United States (Davison 

et al. 2021a). The few acres devoted to chestnut production in the United States are largely 

planted to Chinese chestnut due to its blight resistance (Revord et al. 2021). Agroforestry 

systems have been touted as a means to mitigate the environmental impacts of row crops and 

chestnuts have been proposed as a key component of such systems. If agroforestry systems 

become more widely adopted, plantings of chestnut may expand dramatically, and Darling 58 

American chestnut could be used in agroforestry systems. However, commercial use of Darling 

58 American chestnut is not anticipated at this time for the following reasons: 

• Most growers consider Chinese chestnuts to be the best option currently available for 

establishing profitable orchards in eastern United States (Davison et al. 2021a). Darling 

58 American chestnut was developed as a non-profit project for forest restoration. Its 

attributes have yet to be established and it is susceptible to other diseases such as ink 

disease caused by Phytophthora cinnamomi (ESF 2019). In contrast, Chinese chestnut 

has resistance to ink disease in addition to blight (Metaxas 2013). Furthermore, Chinese 

chestnut has been cultivated for over 2000 years allowing for the selection of favorable 

quality traits. It has been cultivated and bred in the United States since 1930 (Metaxas 

2013). From the standpoint of performance characteristics, the risk of using Darling 58

American chestnut in a commercial venture is much higher than Chinese chestnut.

• Varieties created through genetic engineering typically have less public acceptance than

conventional varieties. Darling 58 American chestnut would pose a risk to commercial 

growers from the standpoint of potential reduced public acceptance. 
vi

• Chinese chestnuts are smaller trees and more suitable for orchard planting than American 

chestnut

• Market surveys suggest American consumers prefer larger chestnuts (Aguilar et al. 2009)

and American chestnuts are smaller than Chinese and Japanese chestnuts. Chinese 

chestnuts are also easier to peel (Metaxas 2013; Davison et al. 2021b). 

In the early 1900s American chestnut was a valued timber tree (Wang et al. 2013) making up 

more than 25 percent of all timber cut in the Southern Appalachians and almost half of the 

timber cut in Connecticut (Hawley and Hawes 1918b; Hepting 1974; Wang et al. 2013). Most 

hardwood timberland forests are regenerated through natural means (i.e, seedlings in the 

understory are given the opportunity to grow upon the selective removal or clear cutting of larger 

trees) (Mississippi State Extension NA). While Darling 58 American chestnut may be able to 

establish and colonize much of the eastern United States if it shows enhanced tolerance to the 

fungus C. parasitica, it may require centuries before becoming a significant enough presence in 

the landscape to allow for timber harvest. The rate of increase will depend on the degree of 

human assistance. Darling 58 American chestnut was developed as a non-profit project for forest 

restoration in American chestnut’s native range. If landowners have a strong interest in American 

chestnut restoration or the perception that Darling 58 American chestnut will provide more value 

to their forest than their existing tree stand and Darling 58 American chestnut seedlings become 

available in large numbers, the increase could be accelerated. As the climate and ecology of the 

eastern forests have changed in the last hundred years it is unknown whether Darling 58 

American chestnuts will ever regain the dominance it exhibited in the nineteenth century.

Darling 58 American chestnut’s attributes have yet to be established for timber production and so 

the risk of using Darling 58 American chestnut in a commercial venture is likely too high for 

adoption in the foreseeable future.

No socioeconomic impacts are expected from deregulation of Darling 58 American chestnut for 

several reasons. As noted above, Darling 58 American chestnut is not expected to be used in 

commercial plantings for timber or nut production for the foreseeable future. The draft EIS 

considered whether Darling 58 American chestnut could impact commercial plantings of 

chestnut via cross pollination, especially those chestnuts produced for a biotech sensitive market. 

Gene flow from Darling 58 American chestnut to commercial chestnut is considered unlikely for 

several reasons. Darling 58 American chestnut is most likely to be planted in the native

American chestnut range while the majority of commercial chestnut production (58%) occurs in 

states outside the native range (USDA-NASS 2019). Darling 58 American chestnut is a different 

species than the chestnut used for commercial production and so, successful hybridization occurs 

at a lower frequency than within the species (ESF 2019). Chinese chestnut also usually flowers 

earlier than American chestnut which decreases effective cross-pollination (Pennsylvania 

Chapter The American Chestnut Foundation 2006). And finally, chestnut pollen does not travel 

long distances. Effective pollination does not occur beyond 400 m (Forest et al. 1977; Russell 

1987; Rutter 1990) so there is likely to be adequate isolation distances between Darling 58

American chestnut and chestnut orchards catering to the biotech sensitive markets. Chestnut

farms catering to the biotech sensitive markets are likely to be low input operations catering 

directly to the consumer (“uPick” operations) which would not be subject to commingling in a 

supply chain (Gullickson 2019). Organic chestnut production today only represents 0.33% of the 
vii

chestnuts sold in the United States (Davison et al. 2021a) though it is unknown whether organic 

chestnut production would capture additional market share if the industry dramatically expanded.

More detailed descriptions and analyses of the potential environmental consequences can be 

found in Chapter 4.
viii

TABLE OF CONTENTS

1 PURPOSE AND NEED FOR AGENCY ACTION................................................1-1

1.1 State University of New York College of Environmental Science and Forestry (ESF) 

Petition for Nonregulated Status.......................................................................1-1

1.2 Purpose of Darling 58 American Chestnut .......................................................1-1

1.3 Coordinated Framework Review and Regulatory Review................................1-2

1.3.1 USDA-APHIS............................................................................................1-2

1.3.2 Environmental Protection Agency.............................................................1-2

1.3.3 Food and Drug Administration ..................................................................1-3

1.4 Purpose and Need for APHIS Action................................................................1-3

2 SCOPING AND PUBLIC INVOLVEMENT .........................................................2-1

2.1.1 Public Comments for Petition 19-309-01p ................................................2-1

2.1.2 Public Scoping for this draft EIS ...............................................................2-1

2.2 Issues Considered in this EIS............................................................................2-2

3 ALTERNATIVES....................................................................................................3-3

3.1 No Action Alternative: Continue as Regulated.................................................3-3

3.2 Preferred Alternative: Determination that Darling 58 American Chestnut is No Longer 

Regulated ..........................................................................................................3-3

3.3 Alternatives Considered but Dismissed from Detailed Analysis......................3-4

3.4 Comparison of Alternatives ..............................................................................3-4

4 AFFECTED ENVIRONMENT AND ENVIRONMENTAL CONSEQUENCES4-15

4.1 Scope of Analysis............................................................................................4-15

4.2 Action Area .....................................................................................................4-16

4.2.1 Previous Distribution ...............................................................................4-16

4.2.2 Current Distribution and Blight ...............................................................4-17

4.2.3 Potential Future Distribution....................................................................4-18

4.3 Physical Environment .....................................................................................4-19

4.3.1 Soil Quality ..............................................................................................4-19

4.3.2 Water Resources ......................................................................................4-20

4.3.3 Air Quality and Climate Change..............................................................4-21

4.4 Biological Resources.......................................................................................4-23

4.4.1 Animal Communities...............................................................................4-23

4.4.2 Plant Communities...................................................................................4-25
ix

4.4.3 Gene Flow and Weediness.......................................................................4-27

4.4.4 Biodiversity..............................................................................................4-29

4.5 Human Health .................................................................................................4-30

4.6 Animal Feed ....................................................................................................4-32

4.7 Socioeconomics...............................................................................................4-33

4.7.1 Domestic Economic Environment...........................................................4-33

4.7.2 Trade Economic Environment .................................................................4-36

4.8 Compliance with Federal and State Laws and Regulations, Executive Orders, Policies, 

and Treaties.....................................................................................................4-37

4.8.1 Federal Laws and Regulations.................................................................4-37

4.8.2 National Environmental Policy Act (NEPA)...........................................4-37

4.8.3 Compliance with the Clean Air Act, Clean Water Act, and Safe Drinking 

Water Act.................................................................................................4-38

4.8.4 Executive Orders Related to Domestic Issues.........................................4-38

4.9 Conclusions: Potential Impacts on the Human Environment..........................4-40

4.9.1 Adverse environmental effects that cannot be avoided should the proposal 

be implemented........................................................................................4-40

4.9.2 The relationship between short-term uses of man’s environment and the 

maintenance and enhancement of long-term productivity. .....................4-41

4.9.3 Irreversible or irretrievable commitments of resources that would be 

involved in the proposal should it be implemented.................................4-41

4.9.4 Whether the action would violate or conflict with a federal or state laws or 

local requirements governing protection of the environment..................4-42

4.9.5 Possible conflicts between the proposed action and the objectives of 

federal, regional, state, tribal, and local land use plans, policies, and 

controls for the area concerned................................................................4-42

4.9.6 Energy requirements and conservation potential of various alternatives and 

mitigation measures.................................................................................4-43

4.9.7 Natural or depletable resource requirements and conservation potential of 

various alternatives and mitigation measures..........................................4-44

4.9.8 Urban quality, historic and cultural resources, and the design of the built 

environment, including the reuse and conservation potential of various 

alternatives and mitigation measures.......................................................4-44

4.9.9 Means to mitigate adverse environmental impacts..................................4-44

4.9.10 Economic and technical considerations, including the economic benefits of 

the proposed action..................................................................................4-45
x

4.9.11 The degree to which the action may adversely affect the endangered or 

threatened species or its habitat that has been determined to be critical 

under the Endangered Species Act of 1973.............................................4-45

4.9.12 The degree to which the proposed action affects public health or safety.4-45

4.9.13 Whether the affected environment includes reasonably foreseeable 

environmental trends and planned actions in the affected areas. ............4-45

APPENDIX 1 THREATENED AND ENDANGERED SPECIES .........................4-47

Requirements for Federal Agencies..........................................................................4-47

Potential Effects of Darling 58 American Chestnut on T&E Species.......................4-49

Threatened and Endangered Plant Species and Critical Habitat ............................... 4-50

Threatened and Endangered Animal Species and Critical Habitat........................4-51

Summary of Effects and Determination....................................................................4-53

APPENDIX 2 LIST OF PREPARERS ....................................................................... 62

APPENDIX 3 DISTRIBUTION LIST FOR THIS EIS.............................................. 63

APPENDIX 4 REFERENCES.................................................................................... 63

5 INDEX...................................................................................................................... 75

LIST OF TABLES

Table 1. Summary of Issues of Potential Impacts and Consequences of Alternatives................3-4

Table 2. List of T&E Species within American Chestnut Historic Range ................................4-54

LIST OF FIGURES

Figure 1. Natural range of American chestnut. (Jacobs 2007) .................................................4-16

Figure 2. Sources of U.S. Greenhouse Gas Emissions..............................................................4-23
xi

ACRONYMS AND ABBREVIATIONS

AOSCA Association of Official Seed Certifying Agencies

APHIS Animal and Plant Health Inspection Service

CAA Clean Air Act

CFR Code of Federal Regulations (United States)

CH4 methane

CO2 carbon dioxide

DNA deoxyribonucleic acid

EIS environmental impact statement

EO Executive Order

EPA U.S. Environmental Protection Agency

ESA Endangered Species Act of 1973

FDA U.S. Food and Drug Administration 

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

GHG greenhouse gas

IPPC International Plant Protection Convention

N2O nitrous oxide

NAAQS National Ambient Air Quality Standards

NEPA National Environmental Policy Act of 1969 and subsequent amendments

NHPA National Historic Preservation Act

NMFS National Marine Fisheries Service

NPS Agricultural non-point source

OECD Organization for Economic Cooperation and Development

PIP plant incorporated protectants

PPRA Plant Pest Risk Assessment

PPA Plant Protection Act

T&E threatened and endangered 

USDA U.S. Department of Agriculture

USDA-NASS U.S. Department of Agriculture-National Agricultural Statistics Service

USC United States Code

USFWS U.S. Fish & Wildlife Service

WPS Worker Protection Standard for Agricultural Pesticides
1-1

1 PURPOSE AND NEED FOR AGENCY ACTION

This document is intended to ensure compliance with the National Environmental Policy Act 

(NEPA). NEPA requires agencies to prepare an environmental impact statement (EIS) to be 

included in “every recommendation or report on proposals for legislation and other major 

Federal actions significantly affecting the quality of the human environment” (42 U.S.C. 

§4332(2)(C)).

The United States Department of Agriculture (USDA) Animal and Plant Health Inspection 

Service (APHIS) is currently engaged in decisionmaking relevant to its statutory authority to 

regulate Darling 58 American chestnut under the plant pest provisions of the Plant Protection Act 

(PPA). The Agency has determined that there are possible environmental impacts, as described 

in chapter 4, associated with whatever regulatory decision it renders. Therefore, this document 

has been prepared as part of this APHIS decisionmaking process.

1.1 State University of New York College of Environmental Science and Forestry 

(ESF) Petition for Nonregulated Status

In November 2019, APHIS received a petition from ESF requesting a determination of the 

regulated status of Darling 58 American chestnut. The purpose of the petition was to request 

nonregulated status for Darling 58 American chestnut, and any progeny derived from them to no 

longer be considered regulated under Title 7 of the Code of Federal Regulations part 340 (7 CFR 

part 340). Darling 58 American chestnuts have been engineered to be tolerant to the fungal 

pathogen, Cryphonectria parasitica (ESF 2019). Darling 58 American chestnut is currently 

regulated by APHIS.

APHIS prepared a plant pest risk assessment (PPRA) and this EIS to respond to the request and 

avoid the inappropriate use of public resources regulating a product developed using genetic 

engineering if the agency determines that it has no authority to do so; the need for this action.

1.2 Purpose of Darling 58 American Chestnut

The American chestnut, once one of the most abundant trees within its range in the eastern 

United States, was a fast-growing and long-lived canopy tree that produced a consistent crop of 

healthful nuts, was harvested for valuable lumber, and was considered a keystone species for 

wildlife. That ended when an invasive fungal pathogen, C. parasitica, was introduced from Asia 

and killed over 3 billion American chestnuts throughout their natural range.

Darling 58 American chestnut was developed using genetic engineering to be tolerant to the 

fungal pathogen, C. parasitica, chestnut blight (ESF 2019). Tolerance to chestnut blight in 

Darling 58 American chestnut was developed by the insertion of an oxalate oxidase (OxO) gene 

from wheat into an American chestnut line known as Ellis (ESF 2019). Tolerance to chestnut 

blight is achieved by detoxifying the oxalic acid produced by the fungus, preventing the acid 

from killing the chestnut’s tissues. In the presence of OxO, the damage caused by the oxalic acid 

is restricted to superficial cankers (ESF 2019).
1-2

1.3 Coordinated Framework Review and Regulatory Review

Since 1986, the U.S. government has regulated organisms developed using genetic engineering 

pursuant to a regulatory framework known as the Coordinated Framework for the Regulation of 

Biotechnology (referred to as the Coordinated Framework). The Coordinated Framework, 

published by the White House Office of Science and Technology Policy, describes the regulatory 

roles and authorities for the three major agencies involved in regulating organisms developed 

using genetic engineering: the USDA APHIS, the U.S. Environmental Protection Agency (EPA),

and the U.S. Food and Drug Administration (FDA). On January 4, 2017, the USDA, EPA, and 

FDA released a 2017 update to the Coordinated Framework (USDA-APHIS 2018), and the 

accompanying National Strategy for Modernizing the Regulatory System for Biotechnology 

Products (ETIPCC 2017). A more detailed description can be found in the original 1986 policy 

statement (51 FR 23302) and in the 2017 Coordinated Framework update (US-EPA 2017).

1.3.1 USDA-APHIS

APHIS regulations at 7 CFR part 340, which were promulgated pursuant to the Plant Protection 

Act (PPA), as amended (7 U.S. Code (U.S.C.) 7701–7772), govern the introduction (importation, 

interstate movement, and environmental release) of organisms developed using genetic 

engineering that may pose a plant pest risk. An organism developed using genetic engineering is 

regulated under 7 CFR part 340 when APHIS has reason to believe that the organism may be a 

plant pest or APHIS does not have sufficient information to determine if the organism is unlikely 

to pose a plant pest risk. An organism is no longer subject to the plant pest provisions of the PPA 

or to the regulatory requirements of 7 CFR part 340 when APHIS determines that the organism is 

unlikely to pose a plant pest risk.

1.3.2 Environmental Protection Agency

The EPA is responsible for regulating the sale, distribution, and use of pesticides, including 

pesticides that are produced by organisms developed using genetic engineering, termed plant 

incorporated protectants. The EPA regulates pesticides under the Federal Insecticide, Fungicide, 

and Rodenticide Act (FIFRA) (7 U.S.C. 136 et seq.) and certain microorganisms under the Toxic 

Substances Control Act (TSCA) (15 U.S.C. 53 et seq.). Before a pesticide is legally used in the 

United States, the EPA must evaluate the pesticide to ensure that it will not cause unreasonable 

adverse effects on the environment and a reasonable certainty of no harm to humans when used 

in accordance with label instructions. Pesticides that complete this evaluation are issued a 

“registration” that permits their sale and use according to requirements set by the EPA. The EPA 

must also approve the language used on the pesticide label in accordance with 40 CFR part 158. 

Once registered, a pesticide may not legally be used unless the use is consistent with the 

approved directions for use on the pesticide's label. The overall intent of the label is to provide 

clear directions for effective product performance while minimizing risks to human health and 

the environment. Under FIFRA the EPA has a standard of reviewing pesticide registrations every 

15 years (US-EPA 2011). The Food Quality Protection Act (FQPA) of 1996 amended FIFRA, 

and set a standard to reassess, over a 10-year period, all pesticide tolerances that were in place 

when the FQPA was signed, make a safety finding when setting tolerances that the pesticide can 

be used with “a reasonable certainty of no harm,” take into consideration aggregate and 
1-3

cumulative effects/risks in assessing human health, and emphasize risks to special subpopulations such as infants and children (US-EPA 2015). 

The EPA also sets tolerances (maximum limits) for pesticide residues that may remain on or in 

food and animal feed, or establishes an exemption from the requirement for a tolerance, under 

the Federal Food, Drug, and Cosmetic Act (FFDCA; 21 U.S.C. 301 et seq.). In establishing a 

pesticide tolerance, the EPA conducts dietary risk assessments to ensure that all tolerances 

established for each pesticide and food product reach a safety determination based on a finding 

of reasonable certainty of no harm. The FDA enforce pesticide tolerances set by the EPA to 

ensure the safety of the nation's food supply.

1.3.3 Food and Drug Administration

The FDA regulates organisms developed using genetic engineering under the authority of the 

FFDCA (21 U.S.C. 301 et seq.). The FDA published its policy statement concerning regulation 

of products derived from new plant varieties, including those derived from genetic engineering, 

in the Federal Register on May 29, 1992 (57 FR 22984). Under this policy, the FDA implements 

a voluntary consultation process to ensure that human food and animal feed safety issues or other 

regulatory issues, such as labeling, are resolved before commercial distribution of food

developed using genetic engineering. This voluntary consultation process provides a way for 

developers to receive assistance from the FDA in complying with their obligations under Federal 

food safety laws prior to marketing.

More recently, in June 2006, the FDA published recommendations in “Guidance for Industry: 

Recommendations for the Early Food Safety Evaluation of New Non-Pesticidal Proteins 

Produced by New Plant Varieties Intended for Food Use” (US-FDA 2006) for establishing 

voluntary food safety evaluations for new non-pesticidal proteins produced by new plant 

varieties intended to be used as food, including plants developed using genetic engineering. 

Early food safety evaluations help make sure that potential food safety issues related to a new 

protein in a new plant variety are addressed early in development. These evaluations are not 

intended as a replacement for a biotechnology consultation with the FDA, but the information 

may be used later in the biotechnology consultation.

1.4 Purpose and Need for APHIS Action

Under the authority of the plant pest provisions of the Plant Protection Act (7 U.S.C. 7701 et 

seq.), the regulations in 7 CFR part 340, "Movement of Organisms Modified or Produced 

Through Genetic Engineering,” regulate, among other things, the importation, interstate 

movement, or release into the environment of organisms modified or produced through genetic 

engineering that are plant pests or pose a plausible plant pest risk. APHIS recently revised 7 CFR 

part 340 and issued a final rule, published in the Federal Register on May 18, 2020 (85 FR 

29790-29838, Docket No. APHIS-2018-0034).2 The new Regulatory Status Review (RSR) 

process, which replaces the petition for determination of nonregulated status process, became

effective for all crops as of October 1, 2021. The petition for a determination of nonregulated 

status subject of this EIS is being evaluated in accordance with the regulations at 7 CFR 340.6 

(2020) as it was received by APHIS, in November 2019, prior to the implementation of the 

2To view the final rule, go to www.regulations.gov and enter APHIS-2018-0034 in the Search field.
1-4

revised regulation. Pursuant to the terms set forth in 7 CFR 340.6 (2020), any person may submit 

a petition to APHIS seeking a determination that an organism should not be regulated under 7 

CFR part 340. APHIS must respond to petitioners with a decision to approve or deny the 

petition. An organism produced using genetic engineering is no longer subject to the 

requirements of 7 CFR part 340 or the plant pest provisions of the PPA if APHIS determines, 

through conduct of a Plant Pest Risk Assessment (PPRA), that it is unlikely to pose a plant pest 

risk. 

Consistent with the Council of Environmental Quality’s (CEQ) NEPA regulations (40 CFR parts 

1500-1508) and USDA and APHIS NEPA implementing regulations and procedures (7 CFR part 

1b, and 7 CFR part 372), APHIS has prepared this EIS to consider the potential impacts of a 

determination of nonregulated status for Darling 58 American chestnut on the human 

environment.3

3 The human environment includes the natural and physical environment and the relationship of people with that environment. 

When economic or social and natural or physical environmental effects are interrelated, the NEPA analysis may addresses these 

potential impacts as well (40 CFR §1508.14).
2-1

2 Scoping and Public Involvement

APHIS seeks public comment on petitions it receives that request a decision of nonregulated

status for organisms developed using genetic engineering. APHIS does this through a notice 

published in the Federal Register. When the Agency decides to prepare an EIS as part of its 

decisionmaking process for a petition, prior to preparation, it also seeks public comments as part 

of its advance scoping process. Details about the public involvement process for the petition that 

is the subject of this document follows.

2.1.1 Public Comments for Petition 19-309-01p

On August 19, 2020, APHIS announced in the Federal Register that it was making ESF’s 

petition available for public review and comment to help identify potential environmental and 

interrelated economic impacts that APHIS should consider in evaluation of the petition (85 FR 

51008-51009). APHIS accepted written comments on the petition for a period of 60 days, until 

midnight, October 19, 2020. At the end of the comment period APHIS received a total of 4,320

public comments. Of the 4,320 comments received 1,619 comments were in opposition to 

deregulation of Darling 58 American chestnut, 2,698 comments were in favor of deregulation, 

and 3 were out of scope. The issues that were raised in the opposing comments were in regard to 

environmental impacts of the unconfined release of a forest tree developed using genetic 

engineering, impacts to native communities, human health and safety impacts of using a wheat 

gene, the need for long term studies, the potential for chestnut to be more susceptible to chestnut 

blight as well as other diseases, the potential for impacts to organic producers, impacts to trade, 

and general anti-biotech sentiments. The in favor of the petition comments emphasized the 

positive environmental and socio-economic benefits of restoring American chestnut to its 

original range. APHIS evaluated the comments and integrated the concerns raised into this draft 

EIS.

2.1.2 Public Scoping for this draft EIS

As part of its scoping process to identify issues to address in this EIS, APHIS also published a 

Notice of Intent (NOI) to prepare the EIS and sought public input during a 30-day comment 

period (August 6, 2021 to September 7, 2021). The docket received a total of 3,964 public 

comments from 3,967 submitters. Of the 3,964 comments received 3,807 comments were in 

opposition to deregulation of Darling 58 American chestnut, 156 comments were in favor of 

deregulation, and 1 was out of scope. Issues most frequently cited in public comments on the 

NOI included:

• the potential for gene flow from Darling 58 American chestnut to wild relatives

• the potential for Darling 58 American Chestnut to spread and become invasive 

• impacts to wildlife, including pollinators, and threatened and endangered species

• the economic impacts, including impacts on organic and non-GMO producers

• non-target impacts specifically to beneficial fungus, the microbiome, and mycorrhizal 

networks

• impacts on the forest ecosystem

• the potential for Darling 58 American chestnut to be more susceptible to insects or other 

pathogens 
2-2

• human health impacts from consuming nuts as well as potential allergies from pollen and 

the need for FDA clearance

• the potential for Darling 58 American chestnut to not be resistant to blight, act as a 

reservoir for blight, or the evolution of pathogen resistance

• the potential for unintended or off-target mutations and impacts from epigenetic changes

• impacts to tribal communities

• traditional cross breeding should be used to develop blight resistance

The issues discussed in this EIS were developed by considering the public input, including 

public comment received from the Federal Register notice announcing the availability of the 

petition (85 FR 51008-51009), the NOI, as well as issues raised in public comments submitted 

for other NEPA documents for organisms developed using genetic engineering, issues raised in 

lawsuits, and other issues raised by various stakeholders. These issues, including those regarding 

the potential reintroduction of American chestnut and the environmental and food/feed safety of 

plants developed using genetic engineering, were addressed to analyze the potential 

environmental impacts of Darling 58 American chestnut.

2.2 Issues Considered in this EIS

The list of resource areas considered in this draft EIS was developed by APHIS through 

experience in considering issues raised by the public, with specific attention to the issues raised 

in public comments submitted for this petition and EAs and EISs of other organisms developed 

using genetic engineering. The resource areas considered also address issues raised in previous 

and unrelated lawsuits, and issues that have been raised by various stakeholders for this and prior

petitions. The resource areas considered in this draft EIS can be categorized as follows:

Action Area:

• Historic, Present, and Potential Future Range of American chestnut

Environmental Considerations:

• Soil Quality

• Water Resources

• Air Quality and Climate Change

• Animal Communities

• Plant Communities

• Gene Flow and Weediness

• Microorganisms

• Biodiversity

Human Health Considerations:

• Consumer Health

Livestock Health Considerations:

• Animal Feed/Livestock Health

Socioeconomic Considerations:

• Domestic Economic Environment 

• Trade Economic Environment
3-3

3 ALTERNATIVES

This document analyzes the potential environmental consequences of a determination of 

nonregulated status of Darling 58 American chestnut. To respond favorably to a petition for 

nonregulated status, APHIS must determine that Darling 58 American chestnut is unlikely to 

pose a plant pest risk. Based on its PPRA (USDA-APHIS 2020), APHIS has concluded that 

Darling 58 American chestnut is unlikely to pose a plant pest risk. Therefore, in the absence of 

any new information, APHIS must determine that Darling 58 American chestnut is no longer 

subject to 7 CFR part 340.

Two alternatives are evaluated in this draft EIS: (1) No Action: Continuation as Regulated and 

(2) Preferred Alternative: Determination of Nonregulated Status of Darling 58 American 

chestnut. APHIS has assessed the potential for environmental impacts for each alternative in 

Chapter 4 Affected Environment and Environmental Consequences.

3.1 No Action Alternative: Continue as Regulated 

Under the No Action Alternative, APHIS would deny the petition. Darling 58 American chestnut

and progeny derived from Darling 58 American chestnut would continue to be regulated under 

the regulations at 7 CFR part 340. Any introduction of Darling 58 American chestnut would still 

require authorization by APHIS. In addition, measures to ensure physical and reproductive 

confinement of Darling 58 American chestnut would continue to be implemented. APHIS might 

choose this alternative if there were insufficient evidence to demonstrate the lack of plant pest

risk from the unconfined cultivation of Darling 58 American chestnut. 

This alternative is not the Preferred Alternative because APHIS has concluded through a PPRA

that Darling 58 American chestnut is unlikely to pose a plant pest risk (USDA-APHIS 2020). 

Choosing this alternative would not satisfy the purpose and need of making a determination of

plant pest risk status and responding to the petition for nonregulated status.

3.2 Preferred Alternative: Determination that Darling 58 American Chestnut is No 

Longer Regulated 

Under this alternative, Darling 58 American chestnut and progeny derived from it would no 

longer be regulated under the regulations at 7 CFR part 340. APHIS would no longer require 

authorizations for introductions of Darling 58 American chestnut and progeny derived from this 

event. This alternative best meets the purpose and need to respond appropriately to a petition for

nonregulated status based on the requirements in 7 CFR part 340 and the agency’s authority 

under the plant pest provisions of the PPA. Because the agency has concluded that Darling 58 

American chestnut is unlikely to pose a plant pest risk (USDA-APHIS 2020), a determination of 

nonregulated status of Darling 58 American chestnut is a response that is consistent with the 

plant pest provisions of the PPA, the regulations codified in 7 CFR part 340, and the 

biotechnology regulatory policies in the Coordinated Framework.
3-4 

3.3 Alternatives Considered but Dismissed from Detailed Analysis 

APHIS has evaluated several additional alternatives for consideration in previous EAs for 

petitions for nonregulated status. For example, APHIS has considered alternatives that would 

entail approving a petition request in part, mandatory isolation or geographic restriction of plants 

developed using genetic engineering and those developed not using genetic engineering, and 

requirements for testing for the presence of plant material from plants developed using genetic 

engineering in plants that were not developed using genetic engineering. 

Based on the PPRA for Darling 58 American chestnut (USDA-APHIS 2020), experience 

regulating organisms developed using genetic engineering, and broad general experience with 

plant varieties, APHIS determined that Darling 58 American chestnut trees are unlikely to pose a 

plant pest risk. Thus, the imposition of testing, release/planting, and/or isolation requirements on 

Darling 58 American chestnut would be inconsistent with the Agency’s statutory authority under 

the plant pest provisions of the PPA, implementing regulations at 7 CFR part 340, and federal 

regulatory policies embodied in the Coordinated Framework. Because it would be unreasonable 

to evaluate alternatives absent any jurisdiction to implement them, these additional alternatives 

stated above were dismissed from detailed analysis in this EA. 

3.4 Comparison of Alternatives

Table 1 presents a summary of the potential impacts associated with selection of either of the 

alternatives evaluated in this EIS. The impact assessment is presented in Chapter 4 of this EIS. 

Table 1. Summary of Issues of Potential Impacts and Consequences of Alternatives. 

Attribute/Measure Alternative A: No Action Alternative B: Determination of 

Nonregulated Status

Meets Purpose and 

Need and Objectives

No Yes

Management Practices

Action Area for 

Darling 58 American 

chestnut 

The American chestnut was once 

one of the most abundant trees 

within its range in the eastern 

United States. American chestnut 

was found at every elevation from 

sea level to over 5000 ft. from as 

far north as Maine and Ontario, 

Canada to as far south as Georgia 

and Mississippi, covering an 

approximate 200 million acres of 

land (Saucier 1973). As a 

consequence of blight, the 

abundance of chestnut has 

drastically declined; the number 

of live stems today is estimated at 

431 ± 30.2 million, approximately 

If Darling 58 American chestnut 

is granted nonregulated status and 

shows enhanced tolerance to the 

fungus C. parasitica, they may be 

able to establish and colonize 

much of the eastern United States 

where populations persist as 

stunted trees, becoming a selfsustaining forest tree species in its 

native range. Given the 

historically slow spread rates, low 

propagule pressure, and need for 

disturbance to provide sufficient 

light for fast growth, the rate of 

increase would likely be very 

slow. Without aggressive 
3-5 

Attribute/Measure Alternative A: No Action Alternative B: Determination of 

Nonregulated Status

10% of the pre-blight population 

(Dalgleish et al. 2015a). The 

American chestnut has been 

reduced from a dominant 

overstory tree to a small 

understory shrub (Elliott and 

Swank 2008; Dalgleish et al. 

2015b). 

restoration efforts, requiring 

considerable effort and 

coordination at landscape scales, 

it may require centuries before 

American chestnut becomes a 

significant presence in the 

landscape (Gustafson et al. 2017; 

Gustafson et al. 2018). 

Physical Environment

Soil Quality American chestnut commonly 

grows on sandy loams in 

association with other hardwoods 

(Saucier 1973). American 

chestnut trees prefer well-drained, 

sandy, and slightly acidic (i.e. pH 

of 5 to 6) soils, often on slopes 

and ridges (Russell 1987; Wang 

et al. 2013). Forest ecosystem 

processes, including 

decomposition, nutrient cycling, 

and productivity, likely changed 

following chestnut's replacement 

by other species.

No significant differences were 

found in colonization by 

ectomycorrhizal fungi in Darling 

58 American chestnut roots 

compared to non-transgenic 

controls, suggesting that the 

presence or expression of OxO in 

Darling 58 American chestnut 

does not pose risks to native soil 

fungi that are ecologically 

important for American chestnuts 

and other trees (ESF 2019). 

Introducing Darling 58 American 

chestnut back into eastern forests 

is unlikely to have negative 

impacts on soil quality. 

Water Resources All water in forested lands 

contain organic matter, inorganic 

matter, and dissolved gasses 

derived from the environment, 

organisms, and anthropogenic 

activities. The concentrations of 

all these substances, in addition to 

their biological, physical, and 

chemical effects, are the basic 

criteria of water quality (Chang 

2013). Surface and groundwater 

resources are key outputs of 

forests. These water resources are 

essential to ecosystem processes 

and functions across the action 

area. 

Introduction of Darling 58 

American chestnut to eastern 

forests should not impact overall 

water resources in the area. 

American chestnut does not have 

different water requirements than 

those tree species currently found 

in eastern forests. Considering the 

relatively slow re-introduction 

rate and spread of Darling 58 

American chestnuts, it is 

anticipated that impacts to water 

resources will remain relatively 

minimal across the landscape. 
3-6 

Attribute/Measure Alternative A: No Action Alternative B: Determination of 

Nonregulated Status

Air Quality and 

Climate Change

Forests play a role in improving 

local and regional air quality. 

Rates of airborne pollution 

removal vary based on the 

pollutant type, leaf season length, 

and precipitation levels. Trees 

remove air pollution by the 

interception of particulate matter 

on plant surfaces and the 

absorption of gaseous pollutants 

through the leaf stomata (Nowak 

et al. 2014). 

Forests play an important role in 

global climate regulation. In 

2007, U.S. forests absorbed an 

estimated 910 million metric tons 

of carbon dioxide equivalent, an 

amount equal to approximately 13 

percent of the country’s gross 

greenhouse gas emissions from 

industrial and other sources 

(Hanson et al. 2010). Carbon 

sequestration is an important 

ecosystem service provided by 

forests globally and represents a 

driving motivation for 

reforestation and conservation 

efforts worldwide. 

Considering that it may require 

centuries before American 

chestnut becomes a significant 

presence in the landscape again, 

in the short term, Darling 58 

American chestnut is unlikely to 

change the role forests play in 

regulating air quality. 

Because American chestnut is 

fast-growing, long-lived, and 

resistant to decay (Ellison et al. 

2005; de Bruijn et al. 2014), its 

reintroduction could result in 

increased carbon sequestration 

and storage in the form of living 

and dead trees and durable wood 

products, however, the magnitude 

of this effect may be minor 

(Gustafson et al. 2017). 

Biological Resources

Animal Communities Pre-blight, American chestnut 

was described as "the most 

important wildlife plant in the 

eastern United States" (Davis 

2005). American chestnut 

produced a heavy mast crop of 

calorie packed seeds. American 

chestnut disappeared from forests 

before many systematic studies of 

wildlife food habits were 

undertaken (Hill 1992), but we 

know that native mammals such 

as white-tailed deer (Odocoileus 

virginianus), cottontail rabbit 

American chestnut was 

considered a keystone species 

meaning it played a critical role in 

the function of the overall 

ecosystem. If Darling 58 

American chestnuts were to 

become established, it would 

influence ecosystem structure and 

function in areas where they were 

introduced, as American 

chestnuts did prior to the blight 

(Paillet 2002). 
3-7 

Attribute/Measure Alternative A: No Action Alternative B: Determination of 

Nonregulated Status

(Sylvilagus floridanus), and black 

bears (Ursus americanus) all 

consumed chestnuts (Diamond et 

al. 2000; Wang et al. 2013), as 

well as many other vertebrates 

including rodents (Lichti et al. 

2014) and birds including wild 

turkeys (Meleagris gallopavo),

American crow (Corvus 

brachyrhynchos), blue jay 

(Cyanocitta cristata), ruffed 

grouse (Bonasa umbellus), and 

the extinct passenger pigeon 

(Ectopistes migratorius) (Webb 

1986; Russell 1987) and heath 

hen (Hill 1992). Additionally, 

American chestnut provided a 

food source to numerous insect 

species, especially during the 

flowering period.

Since American chestnut 

provided a more stable and more 

abundant source of mast than 

oaks, hickory, and beech species 

(Diamond et al. 2000), if Darling 

58 American chestnuts were to 

become established it could result 

in population increases of the 

species that feed on chestnut (Hill 

1992). An increase in American 

chestnut would also result in a 

gradual decline in some cooccurring tree species. American 

chestnut will likely replace other 

tree species in proportion to their 

abundance, rather than replacing 

a single species or genus 

(Gustafson et al. 2017) and 

therefore those species will still 

be available to animal populations 

that use those trees. 

Oxalate oxidase is a common 

enzyme found in all grains, 

several other crops and food 

products, and many wild plants 

and microbes. Animal 

communities within Eastern 

forests are likely already exposed 

to the OxO gene in Darling 58 

American chestnut. Additionally, 

nutrition analyses have confirmed 

that transgenic chestnuts are not 

nutritionally different than their 

wild-type relatives (ESF 2019). 

No impacts are expected to 

animal communities from 

exposure to the OxO gene in 

Darling 58 American chestnut. 

Plant Communities American chestnut can be found 

in naturalized populations 

throughout the eastern United 

States. The plant communities for 

With an increase in American 

chestnut, some co-occurring tree 

species would gradually decline, 

the most likely being the same 
3-8 

Attribute/Measure Alternative A: No Action Alternative B: Determination of 

Nonregulated Status

American chestnut include all of 

the plants in a particular area, 

including native, introduced, 

desirable, and undesirable plants. 

The plant species in the action 

area may be generally 

characterized as forbes, vines, 

succulents, ferns, grasses, shrubs, 

and trees (BONAP 2020). 

Chestnuts were replaced mainly 

by oak and maple, and to a lesser 

extent, hickory, birch, black 

cherry, and others as dominant 

canopy tree species (Keever 

1953; Stephenson 1986; 

Stephenson et al. 1991; Brewer 

1995). No single species has 

emerged in a dominant role 

across a broad geographic range, 

comparable to the pre-blight 

status of American chestnut.

Plant communities in the action 

area are likely to remain largely

unchanged under the No Action 

Alternative.

trees (oak, maple, hickory, birch, 

and black cherry) that replaced 

American chestnut after chestnut 

blight was introduced. American 

chestnut will likely replace other 

tree species in proportion to their 

abundance, rather than replacing 

a single species or genus 

(Gustafson et al. 2017). Given the 

relatively close overlap between 

the niches of chestnut and oak 

(Keever 1953) competition from 

chestnut would likely affect oaks 

more than other species 

(Gustafson et al. 2017). 

Gene Flow and 

Weediness

American chestnut is primarily 

wind-pollinated (Clapper 1954; 

Johnson 1988) and can outcross 

to other chestnut species, 

including Chinese chestnut (C. 

mollissima), Japanese chestnut 

(C. crenata), European chestnut 

(C. sativa), and chinquapin (C. 

pumila) (Jaynes 1964) to form 

hybrids as well as other Castanea

species where their distributions 

overlap. 

American chestnut can regenerate 

vegetatively by sprouting new 

shoots from the root collar, but 

cannot sprout from roots (Paillet 

Under the Preferred Alternative, 

pollen mediated gene flow from 

Darling 58 American chestnut is 

possible. Darling 58 American 

chestnut is intended to be planted 

in proximity to native American 

chestnut trees with the hope that 

wild trees will flower and crosspollinate to yield blight resistant 

seeds with the intention to

increase the genetic diversity of 

the blight tolerant chestnuts. The 

transgenes from Darling 58 could 

also spread to related species 

through successful pollination 

with at least one transgenic parent 
3-9 

Attribute/Measure Alternative A: No Action Alternative B: Determination of 

Nonregulated Status

1984, 1993), so new shoots will 

be in the immediate location of 

the former tree; natural dispersal 

to a new location can take place 

only by seed. Chestnut dispersal 

is relatively slow without human 

intervention. 

to produce viable offspring (ESF 

2019). Gene flow to different 

species is not as likely because 

flowering times are not as 

synchronized, and trees may not 

be in proximity to Darling 58

American chestnut. 

Chestnut pollen is prone to 

desiccation and loses viability 

within as little as a few hours 

when air dried (Maynard 1991), 

so effective pollination becomes 

increasingly unlikely as pollen 

spreads farther from its source. 

Researchers have noted that trees 

need to be within 100 meters for 

successful pollination (Rutter 

1990; Jacobs et al. 2011), and 

trees 300 m apart are essentially 

reproductively isolated from one 

another (Rutter 1990) 

American chestnut trees spread at 

an average rate of "no more than 

a few kilometers per century" 

(Paillet and Rutter 1989). It may 

take a century or more for blight 

tolerant chestnut trees to become 

dominant after the first pioneer 

trees become established in a 

given area. 

Slow natural colonization rates 

and frequent animal and pest 

pressure on seeds and seedlings 

(Clark et al. 2014), in addition 

with the limitations on pollen 

spread, suggest that chestnuts, 

regardless of type or transgene 

status, will not rapidly invade 

new areas (Cook and Forest 

1978). Areas that are not 

intentionally planted with blight-
3-10

Attribute/Measure Alternative A: No Action Alternative B: Determination of 

Nonregulated Status

tolerant chestnuts will likely 

remain without chestnuts for 

decades or longer (ESF 2019). 

Biodiversity In a forest ecosystem,

biodiversity can be affected by 

several factors including climatic 

and soil conditions, evolution, 

changes in species’ geographical 

ranges, population and 

community processes, and natural 

disturbances or those caused by 

human activities (Carnus et al. 

2006). 

Impacts to biodiversity are 

possible with the planting of 

Darling 58 American chestnut. 

Pre-blight, American chestnut 

was described as "the most 

important wildlife plant in the 

eastern United States" (Davis 

2005), a keystone species that 

played a critical role in the 

function of the overall ecosystem. 

While it is difficult to predict 

what impacts Darling 58 

American chestnut will have on 

forest biodiversity, especially 

since the overall ecosystem has 

changed since American chestnut 

disappeared from the landscape, it 

is reasonable to believe that if 

Darling 58 American chestnut 

shows enhanced tolerance to 

chestnut blight and the trees are 

able to establish and spread, in 

the long term it will have positive 

impacts on increasing the 

biodiversity of animals and 

micro-organisms while 

decreasing the abundance of some 

tree species such as oaks (Paillet 

2002). 

Human and Animal Health

Risk to Human Health Pre-blight American chestnut was 

regularly consumed by people 

living within its historic range.

It is the responsibility of food and 

feed manufacturers to ensure that 

the products they introduce into 

commerce are safe and in 

The transformation process in 

Darling 58 American chestnut do 

not cause nutritional differences 

beyond those already present in 

traditionally-bred chestnuts (ESF 

2019) and is not expected to 

result in adverse human health
3-11

Attribute/Measure Alternative A: No Action Alternative B: Determination of 

Nonregulated Status

compliance with applicable laws 

and regulations. Organisms 

developed using genetic 

engineering for food and feed 

may undergo a voluntary 

consultation process with the 

FDA prior to release onto the 

market (US-FDA 2006). 

effects from direct or indirect 

human contact. 

The Darling 58 American 

chestnut tree does not represent a 

source of new, potentially 

allergenic or anti-nutrient 

proteins; the oxalate oxidase gene 

and protein are commonly found 

in a variety of non-allergenic 

foods (ESF 2019). 

An indirect consequence that may 

result from the introduction of 

Darling 58 American chestnut is 

the potential effects from higher 

rodent populations including 

increased Lyme disease risk to 

humans (Dalgleish and Swihart 

2012). While increased Lyme 

disease risk is possible, it is 

unlikely that this risk would be 

significant.

Risk to Animal Feed Historically American chestnut 

was used as feed for hogs (Davis 

2005). Chestnut has low calories 

with rich minerals, vitamins, and 

monounsaturated fatty acid (Joo 

et al. 2018). Currently, other 

varieties of chestnut are 

sometimes used as animal feed

supplement for pigs (All About 

Feed 2011; Joo et al. 2018), 

young poultry and rabbits (Joo et 

al. 2018), and cattle (All About 

Feed 2011). 

Nutritional analyses have 

confirmed that transgenic 

chestnuts are not nutritionally 

different than their wild-type 

relatives (ESF 2019). No impacts 

are expected to animals from 

exposure to the OxO gene in 

Darling 58 American chestnut if 

used in animal feed. 

Socioeconomic

Domestic Economic 

Environment 

American chestnut was a valued 

timber tree (Wang et al. 2013) 

used for construction lumber, 

shingles, fence posts and rails, 

telephone and telegraph poles, 

Darling 58 American chestnut 

would have the same qualities 

that made American chestnut a 

valued timber and nut tree preblight. The U.S. chestnut industry 
3-12

Attribute/Measure Alternative A: No Action Alternative B: Determination of 

Nonregulated Status

paneling, trim, furniture, coffins, 

interior decoration and firewood 

(Detwiler 1915; Buttrick 1925). 

In the early 1900s, American 

chestnut made up more than 25 

percent of all timber cut in the 

Southern Appalachians, was the 

most valuable tree in southern 

New England, and constituted 

almost half of the timber cut in 

Connecticut (Hawley and Hawes 

1918b; Hepting 1974; Wang et al. 

2013). 

Seven years after discovery of the 

blight in 1904 it was estimated 

that the blight had done $25 

million in damages to the 

chestnut industry (Buttrick 1915). 

The chestnut industry is in its 

infancy in the United States but is 

poised to grow due to increased 

consumer demand for chestnuts, 

increased demand for hardwood, 

and the potential for agroforestry 

to replace row crops as a means 

to mitigate climate change and 

the environmental impacts of 

conventional farming. U.S. 

domestic consumption is low at 

0.1lb per capita but demand still 

greatly exceeds supply such that 

90% of chestnuts consumed in the 

United States are imported.

Currently, the preferred species 

for commercial planting is 

Chinese chestnut because of its 

disease resistance, shorter stature, 

shorter time to nut production, 

and superior nut qualities. 

is expected to grow with other 

chestnut species regardless of the 

Darling 58 American chestnut 

regulatory status. 

Darling 58 American chestnut is 

not expected to be used in 

commercial plantings for timber 

or nut production for the 

foreseeable future. Chinese 

chestnut is preferred for nut 

production due to consumer 

preference for larger nut size and 

being easier to peel (Aguilar et al. 

2009). Additionally, Darling 58 

American chestnut was developed 

as a non-profit project for forest 

restoration. Its attributes have yet 

to be established for timber or nut 

production and so the risk of 

using Darling 58 American 

chestnut in a commercial venture 

is likely too high for adoption in 

the foreseeable future. 

Trade Economic 

Environment

Chestnut production in 2018 was 

2.4 million MT, more than any 

Darling 58 American chestnut is 

not expected to alter the trade
3-13

Attribute/Measure Alternative A: No Action Alternative B: Determination of 

Nonregulated Status

other tree nut except coconut

(Davison et al. 2021a). Most of 

this production is in China. The 

United States does not have a 

significant chestnut industry, less 

than one percent of total world 

production (AgMRC 2021). The 

United States has 1,587 farms 

producing chestnuts on 4,228 

acres (USDA-NASS 2019). It 

should be noted that due to 

chestnut blight these figures are

primarily for Chinese chestnut 

and not for American chestnut. A 

few cultivars in production in the 

US are hybrids between 

American chestnut and Chinese 

chestnut or three-way crosses 

between American Chestnut, 

Chinese Chestnut and Japanese 

Chestnut (Revord et al. 2021). 

economic environment for 

chestnut as it is not expected to be 

used in commercial plantings. 

Most growers consider Chinese 

chestnuts to be the best option 

currently available for 

establishing profitable orchards in 

eastern United States (Davison et 

al. 2021a). Darling 58 American 

chestnut was developed as a nonprofit project for forest 

restoration. Its attributes have yet 

to be established and it is 

susceptible to other diseases such 

as ink disease caused by 

Phytophthora cinnamomi (ESF 

2019). From the standpoint of 

performance characteristics, the 

risk of using Darling 58 in a 

commercial venture is much 

higher than Chinese chestnut. It 

also lacks some of the quality 

attributes that consumers prefer 

(large size and easy to peel)

(Aguilar et al. 2009) and because 

it is created using genetic 

engineering may have less public 

acceptance. 

Darling 58 American chestnut is 

not expected to adversely impact 

the biotech sensitive market for 

chestnut because it is unlikely to 

cross pollinate or commingle with 

commercial chestnuts. 

Darling 58 American chestnut is 

not expected to be grown in 

proximity to chestnut species 

used for commercial purposes. 

The majority of commercial 

chestnut operations are outside 

the native range of American 

chestnut (USDA-NASS 2019)
3-14

Attribute/Measure Alternative A: No Action Alternative B: Determination of 

Nonregulated Status

where Darling 58 American 

chestnut is expected to be planted. 

Commercial chestnut is expected 

to be predominantly a different 

species that is unlikely to overlap 

in flowering time. Effective 

pollination of chestnut occurs 

over relatively short distances 

(within 300 m), so isolation 

distances are manageable. 

Growers who cater to a biotech 

sensitive market are likely to sell 

direct to customers without going 

through a supply chain where 

commingling could happen. 

Other Regulatory Approvals

U.S. In 2021, ESF initiated a

consultation with the FDA on the 

safety of Darling 58 American 

chestnut pursuant to the voluntary 

consultation process for crop

plants developed using genetic 

engineering (21 C.F.R. Parts 192 

and 592). A food and feed safety 

and nutritional assessment of 

Darling 58 was submitted to the 

FDA for review (ESF 2021). 

In 2021, ESF initiated a

consultation with the FDA on the 

safety of Darling 58 American 

chestnut pursuant to the voluntary 

consultation process for crop

plants developed using genetic 

engineering (21 C.F.R. Parts 192 

and 592). A food and feed safety 

and nutritional assessment of 

Darling 58 was submitted to the 

FDA for review (ESF 2021).

Compliance with Other Laws

CWA, CAA, EOs Fully compliant Fully compliant
4-15

4 AFFECTED ENVIRONMENT AND ENVIRONMENTAL 

CONSEQUENCES

4.1 Scope of Analysis

Evaluation of the Potential Impacts of Agency Action

An impact would be any change, beneficial or adverse, from existing (baseline) conditions 

described for the affected environment. Thus, impacts or effects means changes to the human 

environment that could result from approval of the petition, release of Darling 58 American 

chestnut and subsequent recolonization of eastern forests by American chestnut. 

Pursuant to CEQ regulations (40 CFR § 1508.1(g), impacts/effects considered are those that are 

reasonably foreseeable and have a reasonably close causal relationship to the petition decision. 

Impacts/effects may occur soon after the Agency decision or occur later in time. Potential 

impacts/effects include ecological (such as the effects on natural resources and on the 

components and functioning of affected ecosystems), historic, cultural, social, or effects on 

public health. Economic effects, such as those on employment or markets, may also be 

considered. Impacts/effects include those resulting from actions that may have both beneficial 

and detrimental effects, even if on balance the agency believes that the effect will be beneficial

(40 CFR § 1508.1(g)).

In considering whether the effects of the proposed action are significant, agencies are to analyze 

the potentially affected environment, and degree of the effects of the action in relation to the 

affected environment (40 CFR § 1501.3). Agencies should also consider connected actions 

consistent with 40 CFR§ 1501.9(e)(1). The potentially affected environment (summarized 

below) is defined by the area(s) potentially impacted by the proposed action (e.g., national, 

regional, or local), and associated resources (e.g., natural, cultural). In considering the degree of 

the effects, agencies are to consider the following, as appropriate to the proposed action:

• Short- and long-term effects.

• Both beneficial and adverse effects.

• Effects on public health and safety.

• Effects that would violate federal, state, tribal, or local laws protecting the environment.

Potentially Affected Environment

The potential environmental impacts of a forest tree developed using genetic engineering occur 

within the context of a forest’s contribution to environmental change. Potential effects on the 

environment will depend on the success of Darling 58 chestnut to survive and spread over time. 

The former range of American chestnut covered approximately 200 million acres of land 

(Saucier 1973) along the Appalachian range, thus, the scale of potential impacts, should take into 

account the current state of eastern forests as well as the potential future forest systems should 

Darling 58 American chestnut be a successful restoration tree.
4-16

USDA-Forest Service defines a forest as a land cover type that is at least 10 percent stocked by 

single stemmed forest trees of any size that will be at least 4 meters tall at maturity (Wear 2013). 

Another characteristic of forests is that when viewed vertically, there is typically a 25 percent or 

greater canopy cover (Wear 2013). Forests provide valuable services in maintaining optimal air, 

water, and soil quality. Additionally, forests provide resources necessary to maintain animal and 

plant communities. Forest biodiversity is dependent on the intensity of management within the 

forest and is generally greater where forests are more structurally diverse.

Darling 58 American chestnut is intended to be used as a restoration tree to establish and 

colonize much of the eastern United States where stunted trees persist, once again becoming a 

self-sustaining forest tree species within its native range. It is within this context that APHIS 

evaluates the potential impacts of Darling 58 American chestnut on the human environment if 

released for restoration purposes.

4.2 Action Area

4.2.1 Previous Distribution

American chestnuts are a member of the genus Castanea in the plant family Fagaceae. 

Fagaceae contains approximately 900 species worldwide in 8 to 10 genera (Kremer et al. 2012), 

consisting of evergreen and deciduous trees and shrubs. The American chestnut was once one of 

the most abundant trees within its range in the eastern United States. American chestnut was 

found at every elevation from sea level to over 5000 ft. from as far north as Maine and Ontario, 

Canada to as far south as Georgia and Mississippi, covering an approximate 200 million acres of 

land (Saucier 1973).

Figure 1. Natural range of American chestnut. (Jacobs 2007)
4-17

American chestnut was the predominant forest tree species in terms of both stand density and 

stature (Buttrick 1915; Braun 1950). The American chestnut was a fast-growing and long-lived 

canopy tree that produced a consistent crop of nuts, could be harvested for lumber, and was 

considered a keystone species for wildlife. In many areas, especially on mountain slopes in the 

Appalachian range, American chestnut was the dominant forest tree. Reports estimated that 

American chestnut lived several hundred years, up to an age of 400 to 600 years, in the southern 

Appalachians, though trees over 100 years of age, grow hollow in the center while continuing to 

grow in diameter (Zon 1904; Buttrick 1925). American chestnut trees were commonly recorded 

at heights of 70 to 100 feet, with diameters of 3 to 5 feet or more (Detwiler 1915; Buttrick 1925; 

Smith 2000).

4.2.2 Current Distribution and Blight

The range of American chestnut was naturally expanding northwestward at the time of blight 

introduction (Brewer 1995). The American chestnut is now found in the lower peninsula of 

Michigan (Brewer 1995), southwestern Wisconsin (Paillet and Rutter 1989), Illinois (Russell 

1987), Iowa (Russell 1987; Farrar 2001), Louisiana, Missouri, and Florida (Dalgleish et al. 

2015a). The observed expansion is presumably due to naturalized populations from historic 

plantings. Some trees planted in the 1800s in the Pacific Northwest remain alive and blight-free 

today (Gillis 2017).

Commercial chestnut production in the United States extends outside the native range of 

American chestnut (Michigan, California, Iowa, Ohio, and Florida are the top 5 producers 

(USDA-NASS 2019) and largely utilizes a different chestnut species, Chinese chestnut (C. 

mollissima), due to its superior resistance to chestnut blight. Domestic chestnut production also 

includes hybrids between Chinese chestnut and American chestnut (C. dentata) and triple 

hybrids between Chinese, American, and Japanese chestnut (C. crenata) (Revord et al. 2021).

The hybrids have increased resistance to blight compared to American chestnut.

In the first decade of the twentieth century, a fungal canker disease was discovered in the Bronx 

Zoological Park, New York, which was disfiguring and quickly killing American chestnut trees 

(Merkel 1905). The fungus was described by mycologist William Murrill (Murrill 1906)) as 

Diaporthe parasitica, which was soon reclassified Endothia parasitica (Anderson and Anderson 

1912) and later Cryphonectria parasitica (Murrill) Barr. (herein referred to as C. parasitica). In 

the decades following its discovery, the blight spread throughout the American chestnut range 

and killed over 3 billion trees. By 1940 virtually all American chestnut was considered 

functional extinction due to blight (Saucier 1973).

The blight fungus infects the stem through wounds or cracks in the bark and forms cankers on 

the aboveground portions of the plant. The cankers girdle and kill living stems. The pathogen 

kills living tissue primarily by secreting a toxin called oxalic acid (McCarroll and Thor 1978). 

Although the aboveground portion of the plant is killed by the fungus, the plants have the ability 

to sprout from the root collar and lower portion of the stem.

As a consequence of blight, the abundance of chestnut has drastically declined; the number of 

live stems today is estimated at 431 ± 30.2 million, approximately 10% of the pre-blight 

population (Dalgleish et al. 2015a). While data on the size class distribution before the blight is 
4-18

limited, the current population is highly skewed toward small trees with the vast majority of 

stems (estimated 84%) less than 2.5cm diameter at breast height (dbh) (Dalgleish et al. 2015a). 

The American chestnut has been reduced from a dominant overstory tree to a small understory 

shrub (Elliott and Swank 2008; Dalgleish et al. 2015b). Chestnuts were replaced mainly by oak 

and maple, and to a lesser extent, hickory, birch, black cherry, and others as dominant canopy 

tree species (Keever 1953; Stephenson 1986; Stephenson et al. 1991; Brewer 1995). No single 

species has emerged in a dominant role across a broad geographic range, comparable to the preblight status of American chestnut.

Under the No Action Alternative, Darling 58 American chestnut would remain regulated and 

would not be able to be planted without APHIS authorizations. American chestnut would remain 

stumps and small understory shrubs (Elliott and Swank 2008; Dalgleish et al. 2015b) in the 

forests of the eastern United States where populations persist.

4.2.3 Potential Future Distribution

American chestnuts are still found wherever American chestnut was a canopy tree before the 

blight. They persist as stumps and small trees, often multi-stemmed, as a result of the blight, and 

no longer occupy a dominant canopy position (Keever 1953; Woods and Shanks 1959; Good 

1968; Mackey and Sivec 1973; Stephenson 1986; Elliott and Swank 2008; Ireland et al. 2011). If 

Darling 58 American chestnut shows enhanced tolerance to the fungus C. parasitica, they may 

be able to establish and colonize much of the eastern United States where populations persist as 

stunted trees, becoming a self-sustaining forest tree species in its native range. Given the 

historically slow spread rates, low propagule pressure, and need for disturbance to provide 

sufficient light for fast growth, the rate of increase would be very slow. Without aggressive 

restoration efforts, requiring considerable effort and coordination at landscape scales, it may 

require, centuries before American chestnut becomes a significant presence in the landscape

(Gustafson et al. 2017; Gustafson et al. 2018). Despite the slow spread, the action area for this 

EIS are those areas of the United States where American chestnut was once a dominant forest 

tree.

Forest Restoration with American Chestnut 

American chestnut is a fast-growing species with the ability to persist in shaded conditions and it 

responds favorably to forest management techniques that limit competition and increase 

available sunlight (Wang et al. 2013). One model found that an aggressive planting effort would 

be required to restore chestnut to its former prominence and that without this aggressive 

restoration effort, requiring considerable effort and coordination at landscape scales, it could take 

a millennium or more for chestnut to fully occupy its former landscapes (Gustafson et al. 2017). 

American chestnut is a prolific seed producer able to produce fruit as early as age 4 when opengrown, or at about 8 to 20 years old when competing with other trees in the forest (Paillet and 

Rutter 1989). Reproduction starts earlier and yields more nuts in trees reproduced by coppice 

than in trees regenerated from seed (Wang et al. 2013). Chestnuts were historically an important 

food for wildlife, to the point that seed predation combined with insect damage likely made 

natural regeneration from seeds rare pre-blight (Detwiler 1915). As a result of the high demand 

for chestnuts by wildlife and humans, Paillet and Rutter (1989) reported that only 1 to 5 viable 
4-19

seeds per tree germinated into seedlings that survived for more than 1 year. Additionally, small 

seedlings are easily killed by fire or frosts (Wang et al. 2013). As a result, artificial regeneration 

of nursery-grown seedlings will be a crucial component of restoration. 

Seeds may be sown directly in the soil or started in containers after cold stratification. Seedlings 

typically grow rapidly and with good form, supported by the seed’s supply of carbohydrates and 

nutrients (ESF 2019). Once sufficient lines of blight-resistant seedlings are developed and tested, 

they can be planted, cross-pollinate with one another, and sexually reproduce. However, 

restoration of this species will probably always involve planting, due to the loss of seedlings 

from nut predation and high seedling mortality (Wang et al. 2013).

Chestnut is self-infertile, so trees will only produce seed when a sexually compatible, flowering 

tree is close enough to pollinate female flowers. Awareness of effective pollination distance is 

crucial in planning restoration plantings and seed production orchards and will be used by 

managers to predict and control the hybridization of various chestnut species and varieties (ESF 

2019). If Darling 58 American chestnut is granted nonregulated status and shows enhanced 

tolerance to the fungus C. parasitica, the trees may be able to establish and colonize much of the 

eastern United States where stunted trees persist, once again becoming a self-sustaining forest 

tree species within its native range in the long term (centuries). Given the historically slow 

spread rates, low propagule pressure, and need for disturbance to provide enough light for fast 

growth, chestnut is not likely to spread significantly in the short term (decades).

4.3 Physical Environment

4.3.1 Soil Quality

Soil consists of solids (minerals and organic matter), liquids, and gases. This body of inorganic 

and organic matter is home to a wide variety of fungi, bacteria, and arthropods, as well as the 

growth medium for terrestrial plant life (USDA-NRCS 2004). Soil is characterized by its layers 

that can be distinguished from the initial parent material due to additions, losses, transfers, and 

transformations of energy and matter (USDA-NRCS 2010). It is further distinguished by its 

ability to support rooted plants in a natural environment. Soil plays a key role in determining the 

capacity of a site for biomass vigor and production in terms of physical support, air, water, 

temperature moderation, protection from toxins, and nutrient availability. Soils also determine a 

site’s susceptibility to erosion by wind and water, and a site’s flood attenuation capacity. Soil 

health may be monitored as an indicator of overall environmental health. Natural conditions and 

anthropogenic actions, such as soil preparation, planting, cultivating and irrigation, continuously 

affect and determine soil health, which in turn can alter the global environment (Lal 2008).

Soil properties including temperature, pH, soluble salts, the amount of organic matter, the carbon 

nitrogen ratio, the numbers of microorganisms, and soil fauna all vary seasonally, as well as over 

extended periods of time (USDA-NRCS 1999). Soil texture and organic matter levels directly

influence its shear strength, nutrient holding capacity, and permeability. Soil taxonomy was 

established to classify soils according to the relationship between soils and the factors 

responsible for their character (USDA-NRCS 1999). Soils are organized into four levels of 

classification, the highest being the soil order. Soils are differentiated based on characteristics 

such as particle size, texture, and color, and classified taxonomically into soil orders based on 
4-20

observable properties such as organic matter content and degree of soil profile development

(USDA-NRCS 2010). The Natural Resources Conservation Service (NRCS) maintains soil maps 

on a county level for the entire United States and its territories (Palm et al. 2007; USDA-NRCS 

2010). 

Although adapted to a variety of site conditions, American chestnut commonly grows on sandy 

loams in association with other hardwoods (Saucier 1973). American chestnut trees prefer welldrained, sandy, and slightly acidic (i.e. pH of 5 to 6) soils, often on slopes and ridges (Russell 

1987; Wang et al. 2013). Alkaline or limestone-derived soils, or very wet or dry soils, do not 

support chestnut colonization (Paillet 2002).

American chestnut may have affected nutrient cycling and soil chemistry (Ellison et al. 2005).

Forest ecosystem processes, including decomposition, nutrient cycling, and productivity, likely

changed following chestnut's replacement by other species because American chestnut had a 

rapid growth rate and sprouting ability, wood with an extremely high tannin content, and leaves 

with a relatively low carbon to nitrogen ratio which influenced these ecosystem processes

(Ellison et al. 2005). Decomposition of chestnut wood is much slower than other co-occurring 

hardwoods and its high tannin concentrations could restrict the mobilization of nutrients in soils. 

Additionally, chestnut's fast growth rate might have resulted in rapid sequestration of carbon and 

nutrients (Ellison et al. 2005).

Chestnut leaf litter decomposes more rapidly in the first year than oak or cherry leaf litter, and 

soils with chestnut leaf litter were shown to have lower nitrogen leaching rates and greater 

dissolved organic carbon than soils with cherry or oak leaf litter (Schwaner and Kelly 2019). 

These differences represent a potential for increased storage of carbon in surface soil of forests 

with successful introduction of Darling 58 American chestnut as the microbial community 

accumulates biomass in a nitrogen-limiting environment. Studies conducted by the petitioner 

showed there were no significant differences in colonization by ectomycorrhizal fungi in roots 

compared to non-transgenic controls, suggesting that the presence or expression of OxO in 

Darling 58 American chestnut does not pose risks to native soil fungi that are ecologically 

important for American chestnuts and other trees (ESF 2019). Introducing Darling 58 American 

chestnut back into eastern forests is unlikely to have negative impacts on soil quality.

4.3.2 Water Resources 

The principal law governing the nation’s water resources is the Federal Water Pollution Control 

Act of 1972, better known as the Clean Water Act. The Clean Water Act establishes water 

quality standards, permitting requirements, and monitoring to protect water quality. The EPA sets 

the standards for water pollution abatement for all waters of the United States under the programs 

contained in the Clean Water Act, but, in most cases, gives qualified states the authority to issue

and enforce permits. Drinking water is protected under the Safe Drinking Water Act of 1974 

(Public Law 93-523, 42 U.S.C. 300 et seq.) (US-EPA 2012).

Surface water in rivers, streams, creeks, lakes, and reservoirs support everyday life through the 

provision of water for drinking and other public uses, irrigation, and industry (USGS 2015). In 

2010, about 75 percent of the freshwater used in the United States came from surface water 

sources, whereas the other 25 percent originated from groundwater (USGS 2015). Groundwater 
4-21

is water that flows underground and is stored in natural geologic formations called aquifers

(USGS 2015). In the United States, approximately 40 percent of the population depends on 

groundwater for its drinking water supply (USGS 2018). Currently, the largest use of 

groundwater in the United States is irrigation, representing approximately 70 percent of all the 

groundwater pumped each day (USGS 2018).

All water in forested lands contain organic matter, inorganic matter, and dissolved gasses derived 

from the environment, organisms, and anthropogenic activities. The concentrations of all these 

substances, in addition to their biological, physical, and chemical effects, are the basic criteria of 

water quality (Chang 2013). Surface and groundwater resources are key outputs of forests. These 

water resources are essential to ecosystem processes and functions across the action area. 

American chestnut is a relatively drought tolerant species, as suggested by its historical 

dominance on upland sites with well-drained, sandy soils (Wang et al. 2013). During an earlyseason drought in a hardwood forest in Pennsylvania, American chestnut saplings maintained 

higher leaf water potential than several associated oak (Quercus) species that are known for their 

drought tolerance (Abrams et al. 1990). When compared to co-occurring hardwood species

American chestnut seedlings reported high water use efficiency during exposure to drought

(Bauerle et al. 2006).

Drought stress or overwatering was occasionally unintentionally applied during growth chamber, 

greenhouse, and outdoor care of chestnuts by the petitioner, but Darling 58 offspring were not 

observed to respond any differently than non-transgenic relatives to these stresses (ESF 2019). 

Intentional experiments were not conducted by the petitioner on Darling 58 regarding abiotic 

stress tolerance, but numerous anecdotal observations on combined batches of transgenic and 

non-transgenic chestnuts have not shown obvious differences (ESF 2019).

If blight-tolerant American chestnut were to become established as an important canopy tree, it 

would begin to influence ecosystem structure and function in these areas, as it did prior to the 

blight (Paillet 2002). Chestnut leaf litter may alter aquatic ecology. Chestnut leaf litter 

decomposes more rapidly in the first year than oak or cherry leaf litter, and soils with chestnut 

leaf litter were shown to have lower N leaching rates and greater dissolved organic carbon than 

soils with cherry or oak leaf litter (Schwaner and Kelly 2019), potentially impacting water 

quality through a change in nutrient levels.

Introduction of Darling 58 American chestnut to eastern forests should not impact overall water 

resources in the area. As noted above, American chestnut does not have different water 

requirements than those tree species currently found in eastern forests. If American chestnut 

were to become established throughout its previous range, it is anticipated that the impacts to 

water quality would be similar to what it was prior to the blight (Paillet 2002), altering aquatic 

ecology and changing nutrient levels. These long-term impacts would be dependent on the 

location and density of American chestnut trees.

4.3.3 Air Quality and Climate Change

The Clean Air Act (CAA) requires the maintenance of National Ambient Air Quality Standards 

(NAAQS). The NAAQS, developed by the EPA to protect public health, establishes limits for 
4-22

six criteria pollutants: ozone, nitrogen dioxide, carbon monoxide, sulfur dioxide, lead, and 

Particulate Matter (US-EPA 2020a). The CAA requires states to achieve and maintain the 

NAAQS within their borders. Each state may adopt requirements stricter than those of the 

national standard and each is required by the EPA to develop a State Implementation Plan that 

contains strategies to achieve and maintain the national standard of air quality within the state. 

Areas that violate air quality standards are designated as non-attainment areas for the relevant 

pollutants, whereas areas that comply with air quality standards are designated as attainment 

areas (US-EPA 2020b). 

Forests play a role in improving local and regional air quality. Trees can absorb or trap nitrogen 

dioxide, sulfur dioxide, and particulate matter 10 microns or less in size (Hanson et al. 2010). 

Rates of airborne pollution removal vary based on the pollutant type, leaf season length, and 

precipitation levels. Trees remove air pollution by the interception of particulate matter on plant 

surfaces and the absorption of gaseous pollutants (ozone, nitrogen dioxide, and sulfur dioxide)

through the leaf stomata (Nowak et al. 2014). In 2010, trees and forests in the conterminous 

United States are estimated to have removed 17.4 million tonnes (t) of air pollution, with the 

ozone and nitrogen dioxide accounting for the greatest amount of pollution removal (Nowak et 

al. 2014). The impacts from introducing Darling 58 American chestnut into eastern forests is 

unlikely to change the role forests play in regulating air quality.

Climate change represents a statistical change in global climate conditions, including shifts in the 

frequency of extreme weather (Rosenzweig et al. 2001). The EPA has identified carbon dioxide 

(CO2), methane (CH4), and nitrous oxide (N2O) as the key greenhouse gases (GHG) contributing 

to climate change. Greenhouse gases, including CO2, CH4, and N2O, function as retainers of 

solar radiation (Aneja et al. 2009). While each of these gases occurs naturally in the atmosphere, 

human activity has significantly increased the concentrations of these gases since the beginning 

of the industrial revolution. The primary sources of GHG emissions in the United States are: 

Transportation (29%), Electricity production (25%), Industry (23%), Commercial and 

Residential (13%), and Agriculture (10%). In 2019, U.S. greenhouse gas emissions totaled 6,558

million metric tons of CO2 equivalent (CO2–eq), or 5,746 million metric tons of CO2-eq after 

accounting for sequestration from the land sector (US-EPA 2021). 
4-23

Figure 2. Sources of U.S. Greenhouse Gas Emissions

Source: (US-EPA 2021)

Forests play an important role in global climate regulation. In 2007, U.S. forests absorbed an 

estimated 910 million metric tons of carbon dioxide equivalent, an amount equal to 

approximately 13 percent of the country’s gross greenhouse gas emissions from industrial and 

other sources (Hanson et al. 2010). Carbon sequestration is an important ecosystem service 

provided by forests globally and represents a driving motivation for reforestation and 

conservation efforts worldwide.

Because American chestnut is fast-growing, long-lived, and resistant to decay (Ellison et al. 

2005; de Bruijn et al. 2014), its reintroduction could result in increased carbon sequestration and 

storage in the form of living and dead trees and durable wood products (Gustafson et al. 2017). 

Given relatively low decay rate of chestnut wood (de Bruijn et al. 2014), this carbon also would 

be expected to remain in storage for a longer period (Gustafson et al. 2017). Darling 58 

American chestnut may provide rapid sequestration of carbon and nutrients, increasing long term 

carbon storage, especially when used in afforestation (Ellison et al. 2005; Gustafson et al. 2017).

4.4 Biological Resources

This section provides a summary of the biological environment and includes an overview of 

animals, plants, gene transfer, weeds and weediness, and biodiversity, as well as an assessment 

of the potential impacts to plant and animal communities and the potential for gene movement

from a determination of nonregulated status of Darling 58 American chestnut. 

4.4.1 Animal Communities

Animal communities include wildlife species and their habitats. Wildlife refers to both native 

and introduced species of mammals, birds, amphibians, reptiles, invertebrates, fish and shellfish.

Animals that might be exposed to American chestnut would be individuals of species that 

typically inhabit eastern forests or feed on chestnuts. 
4-24

Pre-blight, American chestnut was described as "the most important wildlife plant in the eastern 

United States" (Davis 2005). American chestnut produced a heavy mast crop of calorie packed 

seeds. Chestnut mast production in pre-blight forests was not directly measured, but estimates 

range from 270 kg/ha to 2500 kg/ha (Diamond et al. 2000; Gilland et al. 2012). Annual hard 

mast output is highly variable for most species and usually less than that produced by American 

chestnut, fluctuating between 9.5 kg/ha and 924.5 kg/ha annually for hickory, black oak, 

northern red oak, chestnut oak, white oak and scarlet oak combined (Diamond et al. 2000). 

American chestnut disappeared from forests before many systematic studies of wildlife food 

habits were undertaken (Hill 1992), but we know that native mammals such as white-tailed deer

(Odocoileus virginianus), cottontail rabbit (Sylvilagus floridanus), and black bears (Ursus 

americanus) all consumed chestnuts (Diamond et al. 2000; Wang et al. 2013), as well as many 

other vertebrates including rodents (Lichti et al. 2014) and birds including wild turkeys 

(Meleagris gallopavo), American crow (Corvus brachyrhynchos), blue jay (Cyanocitta cristata), 

ruffed grouse (Bonasa umbellus), and the extinct passenger pigeon (Ectopistes migratorius) 

(Webb 1986; Russell 1987) and heath hen (Hill 1992).

Like many other large-seeded trees, American chestnut seeds were dispersed by wildlife species 

that included birds and squirrels (Sciurus spp.). The American crow, blue jay, wild turkey, ruffed 

grouse and passenger pigeon were major dispersers of large fruit and probably important for 

chestnut dispersal (Webb 1986; Russell 1987). Large mammals such as white-tailed deer or 

black bear may have also played an important role in the dispersal of American chestnut, as the 

bur is thought to be an evolutionary adaptation for hitchhiking on mammalian fur (Wang et al. 

2013).

American chestnut provided a food source to numerous insect species, especially during the 

flowering period. Pollen feeders in the insect orders coleoptera (beetles), lepidoptera (moths) and 

hymenoptera (bees) have been observed visiting chestnut catkins (Clapper 1954; Opler 1978; 

Hasegawa et al. 2015; Tumminello 2016; Zirkle 2017). In addition to these observed 

interactions, chestnut pollen has been shown to be especially nutritious to bumble bees (Bombus 

terrestris) (Tasei and Aupinel 2008). Chestnut trees and seedlings are also known to be a food 

source and subject to damage by native insects including chestnut sawfly (Craesus castaneae) 

and periodical cicadas (Magicicada spp.), as well as non-native insects including Asiatic oak 

weevil (Cyrtepistomus castaneus) and chestnut gall wasp (Dryocosmus kuriphilus) (Cook et al. 

2013; Clark et al. 2014; Clark et al. 2015). There is evidence suggesting some native insect 

species followed the tree to functional extinction (Opler 1978).

Forest leaf litter can be an important food source for invertebrate communities. A study by 

Smock and MacGregor (1988) on leaf litter nutritional quality showed that American chestnut 

leaf litter is of similar quality to that of pignut hickory (Carya glabra), both of which are of 

better nutritional quality than northern red oak leaf litter. In a laboratory experiment, leaf 

shredding stream invertebrates preferred chestnut and hickory leaf litter to oak, and growth rates 

were faster on chestnut and hickory than on oak. Oaks are the most important tree species to 

replace chestnut, with hickory and other species locally important in some areas (Smock and 

MacGregor 1988). In areas where oaks replaced chestnut, the change in leaf litter quality would 

have directly impacted leaf shredding insect populations, as well as downstream impacts such as 

reduction in fine particulate organic matter released as a byproduct of feeding (Smock and 

MacGregor 1988).
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Preliminary laboratory tests of aquatic insect herbivory on various types of deciduous leaves 

indicated that mayfly (Frenesia difficilis) larvae preferred American chestnut leaves to most 

other leaves commonly found in eastern forests. Nine leaf types were studied: American beech 

(Fagus grandifolia), American chestnut, American sycamore (Platanus occidentalis), chestnut 

oak (Quercus montana), combined red and sugar maple (Acer rubrum and Acer saccharum), 

shagbark hickory (Carya ovata), tulip-tree (Liriodendron tulipifera), and white oak (Quercus 

alba). Only maple and shagbark hickory were preferred over American chestnut by mayfly 

larvae (ESF 2019).

Since American chestnut was considered a keystone species, it played a critical role in the 

function of the overall ecosystem. The loss of American chestnut and changes in food 

availability had far reaching effects, and may have contributed to more unstable, less resilient 

community dynamics (Kelly et al. 2008; Dalgleish and Swihart 2012). If Darling 58 American 

chestnuts were to become established, it would influence ecosystem structure and function in 

areas where they were introduced (Paillet 2002). Since American chestnut provided a more 

stable and more abundant source of mast than oaks, hickory, and beech species (Diamond et al. 

2000), if Darling 58 American chestnuts were to become established it could result in population 

increases of the species that feed on chestnut (Hill 1992). The increase would be most 

pronounced during years of low seed production by other masting species, resulting in less 

fluctuation in those species that consume seeds (Dalgleish and Swihart 2012).

Higher populations could increase the pest potential of some species, and human-wildlife 

interactions could increase. Other indirect and complex consequences may result from changes 

in animal populations such as rodents. Higher rodent populations could potentially lead to 

increased pressure on songbirds as generalist predator populations increase, reduced gypsy moth 

(Lymantria dispar) outbreaks (moth pupae are eaten by mice), and increased Lyme disease risk 

to humans (Dalgleish and Swihart 2012).

The replacement of oak by Darling 58 American chestnut may increase macroinvertebrate 

activity, with potential consequences on population, community, and ecosystem levels since 

chestnut leaf litter is of higher nutritional value for aquatic macroinvertebrates than oak (Smock 

and MacGregor 1988). Additionally, slow decaying chestnut wood may increase stream channel 

complexity as it replaces faster decaying species, providing additional habitat for fish and 

invertebrates (Ellison et al. 2005).

Animal communities within Eastern forests are likely already exposed to the Oxalate oxidase 

gene in Darling 58 American chestnut. Oxalate oxidase is a common enzyme found in all grains, 

several other crops and food products, and many wild plants and microbes. A list of cultivated 

and wild plants containing the OxO gene can be found in Tables 4.2a and 4.2b in the petition 

(ESF 2019). Additionally, nutrition analyses have confirmed that transgenic chestnuts are not 

nutritionally different than their wild-type relatives (ESF 2019). No impacts are expected to 

animal communities from exposure to the OxO gene in Darling 58 American chestnut.

4.4.2 Plant Communities

As noted in Section 4.2 – Action Area, American chestnut can be found in naturalized 

populations throughout the eastern United States. The plant communities for American chestnut 
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include all of the plants in a particular area, including native, introduced, desirable, and 

undesirable plants. The plant species in the action area represents a diverse variety of plant 

species, ranging from understory grasses to overstory trees. 

Plant species in a particular area may be generally characterized as forbes, vines, succulents, 

ferns, grasses, shrubs, and trees (BONAP 2020). The plant classification descriptions below are 

derived from the Biota of North America Project4 (BONAP 2020): 

• Forbes - Herbaceous plants most commonly with relatively broad, usually pinnately 

veined leaves (contrasted with parallel-veined in "grass or grass-like" plants), with all 

perennating or overwintering organs at or below ground level. The forb category, which 

was originally established in an agricultural context to contrast with grass and grass-like 

plants, includes a wide range of herbaceous growth habits, especially if aquatic plants are 

added. Vining, creeping, and trailing herbs also are included within this broad category 

(in the BONAP system). Plants with annual stems becoming woody at the base are 

included as forbs. Primarily herbaceous plants bearing terminal buds at the tips of woody 

caudex branches at or near ground level are referred to the forb category; "cushion 

plants" belong with these. 

• Vines - Perennial plants with woody, above-ground stems that bear overwintering buds 

and do not die back to a basal stem or rhizome in winter. Trees usually have a single 

main stem, are at least 4 meters tall, and have a more or less distinct and elevated crown. 

A few species produce normally short-lived but rapidly growing plants that occasionally 

attain tree-like proportions (e.g., Ricinus communis). 

• Succulents – Plants with stems and leaves that are very soft, fleshy, and often filled with 

juice or sap.

• Ferns - Spore-producing but flowerless and seedless vascular plants that are usually 

differentiated into roots, stems, and leaf-like fronds.

• Grasses - Herbaceous plants with long, narrow, entire, parallel-veined leaves, often 

produced in a basal cluster, with all perennating or overwintering organs below the 

ground. The flowers of these plants usually are reduced in complexity and thus 

inconspicuous. Grasses and grass-like plants include all members of the monocot families 

Cyperaceae, Juncaceae, Juncaginaceae, and Poaceae, some members of the Liliaceae, and 

all members of the pteridophyte family Isoetaceae, but similar leaved-species occurring 

in numerous dicot families were not scored. 

• Shrubs - Perennial plants with woody, above-ground stems that bear overwintering buds 

relatively evenly positioned on the stems and do not die back to a basal stem or rhizome 

in winter. Shrubs are multi-stemmed from the ground, generally attaining a low stature 

(variable in size but usually under 5 meters tall), and producing a poorly-defined crown. 

Some shrubs may be creeping (e.g., Juniperus horizontalis, Gaultheria hispidula); others 

4 Additional information on plant classes can be found on the BONAP website. http://www.bonap.org/ Last 

accessed September, 2020.
4-27

may be "mat-like" or "mound-like" (e.g., Arctostaphylos nevadensis). Various 

exceptional species are also placed here (e.g., Coreopsis gigantea, a "fleshy-stemmed 

shrub"; Coreopsismaritima, a "hollow-stemmed shrub"; Leucanthemum nipponicum, a 

"soft shrub"), and some primarily shrubby species that occasionally reach tree size are 

also characterized as trees

• Trees - Perennial plants with woody, above-ground stems that bear overwintering buds 

and do not die back to a basal stem or rhizome in winter. Trees usually have a single 

main stem, are at least 4 meters tall, and have a more or less distinct and elevated crown. 

A few species produce normally short-lived but rapidly growing plants that occasionally 

attain tree-like proportions (e.g., Ricinus communis).

As noted above in Section 4.2.3, if Darling 58 American chestnut is granted nonregulated status 

and shows enhanced tolerance to the fungus C. parasitica, the trees may be able to establish and 

colonize much of the eastern United States where stunted trees persist, once again becoming a 

self-sustaining forest tree species within its native range. Models have shown that to restore 

American chestnut to its former prominence it would require an aggressive planting effort and 

without that aggressive planting effort it could take a millennium or more for American chestnut 

to fully occupy its former landscapes (Gustafson et al. 2017). 

With an increase in American chestnut, some co-occurring tree species would gradually decline,

the most likely being the same trees that replaced American chestnut after chestnut blight was 

introduced. American chestnut will likely replace other tree species in proportion to their 

abundance, rather than replacing a single species or genus (Gustafson et al. 2017). Given the

relatively close overlap between the niches of chestnut and oak (Keever 1953) competition from 

chestnut would likely affect oaks more than other species (Gustafson et al. 2017). There is no 

reason to believe that the reintroduction of American chestnut would result in reduction of any 

competing tree species to threatened or endangered levels.

4.4.3 Gene Flow and Weediness

Gene flow is a biological process that facilitates the production of hybrid plants, introgression of 

novel alleles, and evolution of new plant genotypes. Gene flow to and from an ecosystem can 

occur on both spatial and temporal scales. In general, plant pollen tends to represent the major 

reproductive method for moving across areas, while both seed and vegetative propagation tend to 

promote the movement of genes across time and space.

American chestnut is primarily wind-pollinated (Clapper 1954; Johnson 1988), though, insects, 

especially bees, likely play a role (Clapper 1954). Multiple bee species have been observed 

visiting catkins on American chestnuts and other chestnut species (Hasegawa et al. 2015; 

Tumminello 2016; Zirkle 2017). Chestnut species are considered self-incompatible (Clapper 

1954; Russell 1987) although there is at least one report that self-fertilization may occur rarely in 

American chestnut, with < 1% to perhaps < 5% of the tree’s flowers (Rutter 1990).

To have high pollination rates, chestnut trees need to be within 30 to 100 m apart and trees 

further than 300 to 400 m apart will generally not pollinate each other (Forest et al. 1977; Russell 

1987; Rutter 1990). Chestnut pollen can travel up to 100 km but effective pollination distances 
4-28

are much shorter due to rapid desiccation as pollen viability decreases with time spent in the air

(Fernandez-Lopez and Alia 2003). 

American chestnut can outcross to other chestnut species, including Chinese chestnut (C. 

mollissima), Japanese chestnut (C. crenata), European chestnut (C. sativa), and chinquapin (C. 

pumila) (Jaynes 1964) to form hybrids. Darling 58 chestnuts have produced offspring through 

controlled pollinations with C. dentata x C. mollissima F1 hybrids, Allegheny chinquapin, and 

European chestnut (ESF 2019). Additionally, hybridization has been shown to occur between 

American chestnut and other Castanea species where their distributions overlap. Although 

chestnut species are sexually compatible, crossing efficiency in the wild between species is less 

efficient than within the species because flowering times often are not coincident (Pennsylvania 

Chapter The American Chestnut Foundation 2006). 

In the central and southern Appalachians, a hybrid population, C. x neglecta, has been described

as plants with intermediate morphology between American chestnut and Allegheny chinquapin; 

(Hardin and Johnson 1985; Johnson 1988). Others have described a taxon called C. x 

alabamensis (Elias 1971), which may be another hybrid, though it has also been considered an 

isolated population of C. ozarkensis (Johnson 1988), or an entirely separate species (Ashe 1923; 

Graves 1950). More recent analyses of C. x alabamensis confirm it is morphologically and 

phylogenetically unique from C. dentata and not likely a hybrid, but leave its species status 

unresolved (Perkins 2016; Perkins et al. 2019). The evolutionary history of North American 

Castanea species is complicated by recent and past hybridization and incomplete lineage sorting, 

and is still not fully understood (Shaw et al. 2012). 

Under the Preferred Alternative, pollen mediated gene flow from Darling 58 American chestnut 

to wild American chestnut populations is intended. To restore American chestnut to the 

landscape, ESF intends to intentionally cross-pollinate Darling 58 American chestnuts with 

surviving remnant American chestnut populations (ESF 2019). The transgenes from Darling 58 

could spread to related species through successful pollination with at least one transgenic parent 

to produce viable offspring (ESF 2019). Chestnut growers have been managing unwanted 

pollination in chestnut orchards since pollen from certain hybrid or interspecific crosses can be 

detrimental to harvests. These same methods for controlling pollination by transgenic chestnuts 

can be applied to potential restoration programs if needed (ESF 2019). 

American chestnut would also likely disperse through the spread of seeds. Chestnut seeds do not 

survive multiple years in natural conditions, so there is no seed bank (Davelos and JAROSZ 

2004). However, rodents, including squirrels, eastern chipmunks (Tamias striatus) and mice 

(Peromyscus spp.), cache seeds for future consumption, including American chestnuts (Toumey 

and Korstain 1947; Lichti et al. 2014) and these seeds may germinate and become established 

seedlings. Blue jays (Darley-Hill and Johnson 1981; Johnson and Webb III 1989), crows (Zon 

1904), and, historically, the now-extinct passenger pigeon (Webb 1986) also likely played a role 

in dispersing American chestnuts. Whole burs are possibly transported by large mammals such 

as black bear and white-tailed deer by hitchhiking on the animals fur (Wang et al. 2013). 

Weediness of American chestnut
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The speed and extent to which a blight-tolerant American chestnut would become established as 

a dominant canopy tree in today’s eastern U.S. forests is difficult to predict. American chestnut 

has the ability to regenerate vegetatively by sprouting new shoots from the root collar, which

allows individual chestnuts to remain alive even in the presence of blight, and allows 

regeneration after fire (Hawley and Hawes 1918b; Toumey and Korstain 1947; Paillet 2002) or 

logging (Buttrick 1915; Faison and Foster 2014). The ability of chestnut trees to generate sprouts 

declines with stem age, but trees over 100 years old may retain the ability to produce sprouts 

(Zon 1904; Russell 1987). Chestnut does not sprout from roots (Paillet 1984, 1993), so new 

shoots will be in the immediate location of the former tree; natural dispersal to a new location 

can take place only by seed.

Since chestnut dispersal is relatively slow, it is likely that at least the first several decades of 

chestnut restoration will depend on people intentionally planting trees and caring for them 

(Gustafson et al. 2017). Chestnut cannot be restored in a short time frame and may require 

considerable effort and coordination at landscape scales (Gustafson et al., 2017).

Areas that are not intentionally planted with blight-tolerant chestnuts will likely remain without 

chestnuts for decades or longer (ESF 2019). Slow natural colonization rates and frequent animal 

and pest pressure on seeds and seedlings (Clark et al. 2014) suggest that chestnuts, regardless of 

type or transgene status, will not rapidly invade new areas (Cook and Forest 1978). A study by 

Paillet and Rutter (1989) of American chestnut trees outside of their natural range showed that 

chestnut trees spread at an average rate of "no more than a few kilometers per century" though 

the rate of spread appeared to be increasing with increased seed production by the established 

trees. American chestnut has the ecological capacity to achieve canopy dominance on favorable 

sites, but that it may take a century or more for blight tolerant chestnut trees to become dominant 

after the first pioneer trees become established in a given area (Paillet and Rutter 1989).

4.4.4 Biodiversity

Biodiversity refers to all plants, animals, and microorganisms interacting in an ecosystem

(Wilson 1988). Biodiversity provides valuable genetic resources for crop improvement and also 

provides other functions beyond food, fiber, fuel, and income (Harlan 1975). The primary 

function of biological diversity is to contribute to ecosystem services. These ecosystem services 

may include: pollination, genetic introgression, biological control, nutrient recycling, 

competition against natural enemies, soil structure, soil and water conservation, disease 

suppression, control of local microclimate, control of local hydrological processes, and 

detoxification of noxious chemicals (Altieri 1999). The loss of biodiversity results in a need for 

costly management practices in order to provide these functions (Altieri 1999).

In a forest ecosystem, biodiversity can be affected by several factors including climatic and soil 

conditions, evolution, changes in species’ geographical ranges, population and community 

processes, and natural disturbances or those caused by human activities (Carnus et al. 2006).

Impacts to biodiversity are possible with the planting of Darling 58 American chestnut. Preblight, American chestnut was described as "the most important wildlife plant in the eastern 

United States" (Davis 2005), playing a critical role in the function of the overall ecosystem. 

American chestnut provided a food source to numerous species. If Darling 58 American chestnut 
4-30

is granted nonregulated status and shows enhanced tolerance to the fungus C. parasitica, the 

trees may be able to establish and colonize much of the eastern United States, influencing 

ecosystem structure and function in areas where they were introduced, as American chestnuts did 

prior to the blight (Paillet 2002). Since American chestnut provided a more stable and more 

abundant source of mast than oaks, hickory, and beech species (Diamond et al. 2000), if Darling 

58 American chestnuts were to become established it could result in population increases of the 

species that feed on chestnut (Hill 1992). The increase would be most pronounced during years 

of low seed production by other masting species, resulting in less fluctuation in those species that 

consume seeds (Dalgleish and Swihart 2012). 

As noted above as American chestnut populations increase, some co-occurring tree species 

would likely gradually decline. American chestnut will likely replace other tree species in 

proportion to their abundance, however, given the relatively close overlap between the niches of 

chestnut and oak (Keever 1953) competition from chestnut would likely affect oaks more than 

other species (Gustafson et al. 2017). The replacement of oak by Darling 58 American chestnut 

may increase macroinvertebrate activity, with potential impacts to population, community, and 

ecosystem levels since chestnut leaf litter is of higher nutritional value for aquatic 

macroinvertebrates than oak (Smock and MacGregor 1988) and slow decaying chestnut wood 

may increase stream channel complexity providing additional habitat for fish and invertebrates 

(Ellison et al. 2005). 

While it is difficult to predict what impacts Darling 58 American chestnut will have on forest 

biodiversity, it is reasonable to believe it will have positive impacts on increasing the 

biodiversity of animals and micro-organisms while decreasing the abundance of some tree 

species such as oaks in the long term. 

4.5 Human Health

Human health considerations associated with plants developed using genetic engineering are 

those related to (1) the safety and nutritional value of the plants and their products to consumers, 

and (2) the potential health effects of pesticides that may be used in association with plants 

developed using genetic engineering. As for food safety, consumer health concerns are in regard 

to the potential toxicity or allergenicity of the introduced genes/proteins, the potential for altered 

levels of existing allergens in plants, or the expression of new antigenic proteins. Consumers 

may also be concerned about the potential consumption of pesticides on foods derived from 

plants developed using genetic engineering. In the case of American chestnut; the chestnuts 

produced could be used for food and feed purposes. 

In the United States, plants developed using genetic engineering and other organisms are 

regulated and evaluated for public health and environmental safety under the Coordinated 

Framework for the Regulation of Biotechnology, described in Section 1.3. The safety assessment 

of plants derived through biotechnology includes characterization of the DNA insert or other 

genetic material, characterization of the biochemical and functional properties of the expressed 

protein(s), and compositional analysis of the plants developed using genetic engineering. 

Food Safety 
4-31

Under the FFDCA and the Food Safety Modernization Act (FSMA), it is the responsibility of 

food and feed manufacturers to ensure that the products they introduce into commerce are safe 

and in compliance with applicable laws and regulations. Organisms developed using genetic 

engineering for food and feed may undergo a voluntary consultation process with the FDA prior 

to release onto the market (US-FDA 2006). Although a voluntary process, thus far all applicants 

who wish to commercialize a variety developed using genetic engineering that will be included 

in the food supply have completed a consultation with the FDA. APHIS considers the voluntary 

FDA assessment in evaluating the potential impacts of a determination on nonregulated status of 

a plant or other organism developed using genetic engineering.

Food safety reviews frequently compare the compositional characteristics of the plant developed 

using genetic engineering with plants not developed using genetic engineering, conventional 

varieties of that plant. Compositional characteristics evaluated in these comparative tests 

typically include plant components such as protein, fat, carbohydrates, ash, minerals, dietary 

fiber, essential and non-essential amino acids, fatty acids, vitamins, and anti-nutrients. Various 

developers have performed characterization analyses of trait genes and proteins, safety 

assessments of the genes and proteins, compositional analyses of food and feed, and safety and 

nutritional assessments of food and feed products derived from plants developed using genetic 

engineering containing these traits (i.e., those submissions listed at (US-FDA 2020; USDAAPHIS 2021). The FDA evaluates the submission and responds to the developer by letter with 

any concerns it may have or additional information it may require. 

ESF plans to consult with the FDA on the safety of Darling 58 American chestnut pursuant to the 

voluntary consultation process for crop plants developed using genetic engineering (21 C.F.R. 

Parts 192 and 592). A food and feed safety and nutritional assessment of Darling 58 will be 

submitted to the FDA for review (ESF 2019).

In addition, foods derived from plants developed using genetic engineering typically undergo a 

safety evaluation among international agencies before entering foreign markets, including 

reviews under Codex Alimentarius guidelines, the European Food Safety Agency (EFSA), and 

Australia and New Zealand Food Standards Agency (ANZFS) (e.g., see (WHO 2005; FAO 2009; 

EFSA 2015)). 

In general, based on over 15 years of peer reviewed research and regulatory review, rather broad 

agreement among the scientific and regulatory communities has emerged that food products 

derived from plants developed using genetic engineering currently on the market are as safe as 

and nutritionally equivalent to their counterparts not developed using genetic engineering, and 

pose no more risks than foods derived from conventional crop varieties (e.g., see (CAST 2005; 

WHO 2005; Batista and Oliveira 2009; Ronald 2011; AAAS 2012; AMA 2012; DeFrancesco 

2013; Goldstein 2014; Nicolia et al. 2014), and reviews by FDA (US-FDA 2020), EFSA (EFSA 

2015), and ANZFS (ANZFS 2015)). 

While the safety of foods derived from current crops developed using genetic engineering has 

been established through peer reviewed research and regulatory agency reviews (e.g., (WHO 

2005; Batista and Oliveira 2009; AAAS 2012; AMA 2012; DeFrancesco 2013; Goldstein 2014), 

and others), some consumers may worry about potential negative health effects from food 

derived from plants developed using genetic engineering; such as through the consumption of 
4-32

introduced DNA, or changes in nutritional quality or allergenicity. Consequently, consumer 

preferences can tend towards avoidance of food derived from plants developed using genetic 

engineering unless such food contains perceptible benefits (Lucht 2015).

The Darling 58 American chestnut tree does not represent a source of new, potentially allergenic 

or anti-nutrient proteins; the oxalate oxidase gene and protein are commonly found in a variety 

of non-allergenic foods (ESF 2019). The OxO enzyme (and its encoding gene) is eaten by over a 

billion people daily in wheat and other grains, and has not been identified as an allergen in any 

known reports (ESF 2019). Additionally, the fact that the OxO enzyme is present in corn, rice, 

sorghum, and many other foods that are not considered allergens (Hefle et al. 1996; FDA 2006)

provides support for a lack of allergenicity in OxO.

Oxalate oxidase and similar enzymes are consumed and handled daily in cereal grains and many 

other foods without any reports of toxicity, and we have found no evidence that it should be 

considered a toxin (ESF 2019).

Under the Preferred Alternative, potential impacts to human health are not anticipated to be 

different from those under the No Action Alternative. The presence of the OxO transgene or the 

transformation process in Darling 58 American chestnut do not cause nutritional differences 

beyond those already present in traditionally-bred chestnuts (ESF 2019) and is not expected to 

result in adverse human health effects from direct or indirect human contact.

Other indirect consequences may result from introduction of Darling 58 American chestnut and 

successful re-establishment of American chestnut to eastern forests. A more stable and more 

abundant source of mast could result in population increases of the species that feed on chestnut 

(Hill 1992) including rodents. Potential effects of higher rodent populations could include 

increased Lyme disease risk to humans (Dalgleish and Swihart 2012). While increased Lyme 

disease risk is possible, it is unlikely that this risk would be significant given the historically 

slow spread rates and that the rate of increase would be very slow. The increased mast 

availability that leads to the increased rodent populations could require centuries.

4.6 Animal Feed

Historically American chestnut was used as feed for hogs (Davis 2005). Unlike other nuts and 

seeds, chestnut has low calories with rich minerals, vitamins, and monounsaturated fatty acid 

(Joo et al. 2018). Currently, other varieties of chestnut not meeting quality standards are used as 

an animal feed supplement for pigs (All About Feed 2011; Joo et al. 2018), have been added to 

the diet of young poultry and rabbit to enhance the growth performance by the improvement of 

gastrointestinal microflora stability (Joo et al. 2018), and have been added to the diets of cattle in 

the last few months of fattening for the market (All About Feed 2011). Additionally tannins 

derived from chestnuts have been used with calves, sheep, and other animals as an alternative to 

antibiotics (Liu et al. 2011a; Liu et al. 2011b; Buccioni et al. 2015; Bonelli et al. 2018).

It is unlikely that American chestnut would be used for animal feed in the near future as chestnut 

dispersal is relatively slow and it will take at least several decades for chestnut restoration to

reach the point of having a surplus nut crop. However, nutritional analyses have confirmed that 

transgenic chestnuts are not nutritionally different than their wild-type relatives (ESF 2019). No 
4-33

impacts are expected to animals from exposure to the OxO gene in Darling 58 American 

chestnut if used in animal feed.

It is the responsibility of feed manufacturers to ensure that the products they market are safe and 

properly labeled. Feed derived from American chestnut developed using genetic engineering 

must comply with all applicable legal and regulatory requirements, which are designed to protect 

human health. To help ensure compliance, a voluntary consultation process with the FDA may 

be implemented before release of commodity products with origins from plants developed using 

genetic engineering as animal feed into the market. 

4.7 Socioeconomics

4.7.1 Domestic Economic Environment

American chestnut was a valued eastern hardwood species due to its use as a timber tree, its 

abundant nut production, and its secondary wood products (Wang et al. 2013). Historically 

American chestnut was used for construction lumber, shingles, fence posts and rails, telephone 

and telegraph poles, paneling, trim, furniture, coffins, interior decoration and firewood (Detwiler 

1915; Buttrick 1925). American chestnut cordwood supplied tanneries one-half of their raw

source of tannin (Buttrick 1925) and was also used in the manufacture of wood pulp (Buttrick 

1925).

In the early 1900s, American chestnut made up more than 25 percent of all timber cut in the 

Southern Appalachians, was the most valuable tree in southern New England, and constituted 

almost half of the timber cut in Connecticut (Hawley and Hawes 1918b; Hepting 1974; Wang et 

al. 2013). The timber yield of American chestnut varied greatly depending on the dominance of 

American chestnut in the stand. Buttrick (1925) estimated that pure stands of American chestnut 

could yield as high as 20,000 board feet per acre while mixed stands would yield an average of 

4,000 board feet per acre. The yield on slopes was estimated at 2,000 to 3,000 board feet per acre 

and that on ridges was reported at 1,500 board feet per acre (Buttrick 1925). Seven years after 

discovery of the blight in 1904 it was estimated that the blight had done $25 million in damages 

to the chestnut industry (Buttrick 1915).

Darling 58 American chestnut would have the same qualities that made American chestnut a 

valued timber and nut tree pre-blight. The Chinese primarily cultivate chestnuts for nut 

production (Qin and Feng 2009) and have selected chestnut trees that grow well in orchards, 

though some varieties of Chinese chestnut were found to have high story canopies suitable for 

timber (Diller 1947; Commender 2017). European chestnut, which grows about the same size as 

American chestnut, is grown for nuts, timber, or both (Pereira-Lorenzo et al. 2009). American 

chestnut, with its vigorous upright growth and open canopy is valued for high quality lumber

(Davison et al. 2021a) and the nuts, while sweet tasting, are less valuable for human 

consumption based on their size. 

Chestnuts in Timber Production

In 2012, 76% of timberland forests in the eastern United States were privately owned (58% by 

non-corporate entities, 24% by corporate entities) and 24% of the timberland forests were on 

public lands (USDA-FS 2014). In the United States, very few hardwood trees are planted in 
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plantations (Zhang and Polyakov 2010). Most hardwood timberland forests are regenerated 

through natural means (i.e, seedlings in the understory are given the opportunity to grow upon 

the selective removal or clear cutting of larger trees) (Mississippi State Extension NA). In cases 

where the seedlings in the understory are undesirable, landowners may incur the expense of 

planting seeds or seedlings to improve the quality of the forest (Mississippi State Extension NA). 

While this is not a common practice for hardwood trees it may become more widely practiced as 

hardwood trees with improved genetics are developed. Darling 58 American chestnut was 

developed as a non-profit project for forest restoration in American chestnut’s native range. 

Darling 58 American chestnut seedlings are intended to be planted in eastern forests. As the 

climate and ecology of the eastern forests have changed in the last hundred years it is unknown 

whether Darling 58 American chestnuts will ever regain the dominance it exhibited in the 

nineteenth century. For example, ink disease, to which Darling 58 American chestnut is 

susceptible has been a problem in the Southern United States (Russell 1987; Gailing and Nelson 

2017) and climate change might expand the problem northward. Nevertheless, the possibility 

remains that if Darling 58 American chestnut is granted nonregulated status and shows enhanced 

tolerance to the fungus C. parasitica, the trees may be able to establish and colonize much of the 

eastern United States where stunted trees persist, once again becoming a self-sustaining forest 

tree species within its native range and potentially providing a valuable source of lumber. Human 

assistance will be essential for the restoration to be successful. However, given the historically 

slow spread rates, low propagule pressure, and need for disturbance to provide enough light for 

fast growth, the rate of increase may require centuries before becoming a significant enough 

presence in the landscape to allow for timber harvest. The rate of increase will also depend on 

the degree of human assistance. If landowners have a strong interest in American chestnut 

restoration or the perception that Darling 58 American chestnut will provide more value to their 

forest than their existing tree stand and Darling 58 American chestnut seedlings become 

available in large numbers, the increase could be accelerated. Pre-blight it was recommended 

that American chestnut be maintained in a rotation of 80 years (Hawley and Hawes 1918a; 

Pinchot 2011) for timber production and it has been noted that “regular and plentiful” nut crops 

are produced after 20 years (Zon 1904; Paillet and Rutter 1989). Darling 58 American chestnut’s 

attributes have yet to be established for timber production and so the risk of using Darling 58 

American chestnut in a commercial venture is likely too high for adoption in the foreseeable 

future. Silviculture for timber production from American chestnut has not had the opportunity to 

advance for over one hundred years and would likely evolve if the restoration is successful. 

Chestnuts in Agroforestry systems

Agroforestry has been touted as a solution to the negative environmental impacts caused by 

monoculture row crops (Davison et al. 2021a). In agroforestry, trees are integrated into the 

agricultural landscape providing direct economic benefits through food, fodder, and lumber, 

climate change mitigation and adaptation through carbon sequestration, and ecosystem services 

such as soil retention, increasing soil water infiltration and storage, reducing evaporation of soil 

moisture, protecting crops from wind stress, stabilizing air and soil temperatures, increasing 

biodiversity, and preventing nutrient runoff (Davison et al. 2021a). Chestnuts, specifically 

Chinese chestnuts, have been proposed as a key tree species in agroforestry systems due to their 

rapid growth, high yield, and consistent production. Agroforestry pilot projects utilizing chestnut 

have been started at both public institutions: University of MO Horticulture and Agroforestry 
4-35

Research5 ; SUNY Adirondack Warren County Conservation District6; North Carolina A and T 

University Four Farms, U NE Horning State Farm Demonstration Forest, and Penn State 

University Rodale Farm7

; and private ventures in New York8 and MO9. If these systems show 

promise, chestnut plantings, but not necessarily American chestnut, are likely to increase. 

Chestnut Nut Production 

There is renewed interest in chestnuts as a food source because the nuts are considered a 

nutritious food. Chestnuts are a source of dietary fiber, contain a significant amount of vitamin 

C, but contain no cholesterol or gluten (Warmund 2011). Unlike other nuts, they are low in fat 

and rich in carbohydrates (Warmund 2011). The nuts could be processed into a gluten free flour 

alternative (Warmund 2011), however chestnut processing is virtually non-existent in the United 

States because demand for fresh chestnuts exceeds the meager supply produced in the United 

States (Davison et al. 2021a). 

Chinese chestnuts have been cultivated and selected for improved varieties for over 2000 years 

in Asia. The chestnut grown in Asia is a different species from American chestnut. As a result of 

the extensive cultivation and selection for nut production, Chinese chestnut has superior qualities 

for commercial nut production. In response to the blight harming American chestnuts, Chinese 

chestnuts, which are resistant to both blight and ink disease caused by Phytophthora cinnamomi

(ESF 2019) have been explored for commercial uses in the United States. An active breeding 

program utilizing Chinese chestnut began in the 1930’s to attempt to adapt varieties to climates 

within the United States as well as to test Asiatic chestnuts for the production of timber (Diller 

1947). The nascent chestnut industry in the United States revolves principally around Chinese 

chestnut (C. mollissima Bl.) though there are some operations using European, Japanese, and 

hybrid chestnuts (Revord et al. 2021). The Agroforestry program at the University of MO Center 

for Agroforestry has been breeding chestnuts and supporting the chestnut industry for 20 years. 

They recently announced a Chestnut Improvement Network, a participatory breeding program 

led by UMCA and partner growers to expand and accelerate efforts in chestnut breeding (Revord 

et al. 2022). Chinese chestnut is the most prevalent species in the breeding program due to its 

superior nut qualities, climatic adaptation, resistance to chestnut blight and phytophthora root rot, 

and shorter stature amenable to orchard production. Most growers consider Chinese chestnuts to 

be the best option currently available for establishing profitable orchards in eastern United States

(Davison et al. 2021a). In contrast, Darling 58 American chestnut’s commercial attributes are 

untested and it remains susceptible to phytophthora root rot, has smaller nuts unfavored by U.S. 

consumers (Aguilar et al. 2009), and is taller than Chinese chestnut making it less desirable for 

nut production in an orchard setting though it may be more suitable for timber (Davison et al. 

2021a). Furthermore, crops created through genetic engineering have less public acceptance 

than conventional crops and as a long-term investment, would also be less likely to be used on 

that basis for nut production. Therefore, despite the promise of an expanding chestnut industry, 

5 FarmHorticulture and Agroforestry Research Farm // Missouri Agricultural Experiment Station 6 Pilot Program To Develop Agroforestry Plan Will Be Implemented At SUNY Adirondack - Glens Falls Business Journal 7 USDA/ 2011. Agroforestry USDA reports to America, Fiscal years 2011-2012-Comprehensive Version 8 Propagate Case Study — Propagate (propagateag.com)), 9 USDA/ 2011. Agroforestry USDA reports to America, Fiscal years 2011-2012-Comprehensive Version: Bringing chestnuts 

back to American landscapes and diets one graft at a time
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Darling 58 American chestnut is not expected to be used in commercial nut operations for the 

foreseeable future.

American chestnut can cross-pollinate with other chestnut species where their distributions 

overlap (Jaynes 1964) potentially leading to impacts to chestnut producers catering to a biotechsensitive market. Currently organic chestnut production only represents 0.33% of the chestnuts 

sold in the United States (Davison et al. 2021a); it is unknown whether organic chestnut 

production would capture additional market share if the industry dramatically expanded. 

American chestnut is primarily wind-pollinated (Clapper 1954; Johnson 1988). To have high 

pollination rates, chestnut trees need to be within 30 to 100 m apart and trees further than 300 to 

400 m apart will generally not pollinate each other (Forest et al. 1977; Russell 1987; Rutter 

1990). Chestnut pollen can travel up to 100 km but effective pollination distances are much 

shorter due to rapid desiccation as pollen viability decreases with time spent in the air

(Fernandez-Lopez and Alia 2003). Managing unwanted pollination of chestnut orchards is 

already an issue that is addressed by chestnut growers, since pollen from certain hybrid or 

interspecific crosses can be detrimental to harvests. Small effective pollination distances for 

chestnut mean that such management is achievable. Controlling pollination by transgenic 

chestnuts after implementation of potential restoration programs would be similarly manageable 

for growers if needed (ESF 2019). There are unlikely to be any impacts to biotech-sensitive 

chestnut producers from a determination of non-regulated status for Darling 58 American 

chestnut for several reasons:

1. As noted above, Darling 58 American chestnut is not expected to be used in commercial 

nut plantings for the foreseeable future. 

2. Darling 58 American chestnut is most likely to be planted in the native American

chestnut range while the majority of commercial chestnut production (58%) occurs in 

states outside the native range (USDA-NASS 2019). 

3. Darling 58 American chestnut is a different species than the chestnut used for 

commercial production. As such, successful hybridization occurs at a lower frequency 

than within the species (ESF 2019). Chinese chestnut also usually flowers earlier than 

American chestnut which decreases effective cross-pollination (Pennsylvania Chapter 

The American Chestnut Foundation 2006).

4. Chestnut pollen does not travel long distances. Effective pollination does not occur 

beyond 400 m so there is likely to be adequate isolation distances between Darling 58

American chestnut and chestnut orchards catering to the biotech-sensitive market.

5. Chestnut farms catering to the biotech-sensitive market are likely to be low input 

operations catering directly to the consumer (“uPick” operations) (Gullickson 2019),

which would not be subject to commingling in a supply chain.

4.7.2 Trade Economic Environment

Worldwide demand for chestnuts exceeds all other nuts except for coconuts and peanuts

(Davison et al. 2021a). China alone produces 1.9 million Metric tons (M MT) of chestnut 

(Modor Intelligence 2022) which is greater than worldwide production of almonds (1.6 M MT), 
4-37

walnuts (1.0 M MT), cashews (0.89 M MT), pistachios (0.80 M MT), or hazelnuts (0.55 M MT) 

(International Nut and Dried Fruit Council 2022). The value of the nut is related to its size. 

Generally, the value of chestnuts ranges from $0.75 to $2.50 per pound wholesale and 

from $2.00 to $5.00 per pound retail, depending on the market (AgMRC 2021). The United 

States does not have a significant chestnut industry, less than one percent of total world 

production, despite being one of the few nations in the world that can grow chestnuts (AgMRC 

2021). The United States has 1,587 farms producing chestnuts on 4,228 acres (USDA-NASS 

2019). The top five states with the most chestnut acreage were Michigan, California, Iowa, Ohio,

and Florida (USDA-NASS 2019). These farms primarily produce chestnuts for fresh market 

domestic consumption. Because demand greatly exceeds supply, about 90% of chestnuts 

consumed in the United States are imported, 3,200 metric tons of chestnuts in 2017 (AgMRC 

2021). The United States has a very low per capita consumption of chestnuts (0.1 lb/person/yr) 

compared to Europe at 1 lb per capita and Korea at 4 lbs per capita. Chestnuts are grown in 27 

countries and worldwide production in 2018 was 2.4 million tons with a value of $5.4 billion

(Davison et al. 2021a). Growth is projected to increase annually by 2.2% over the next five years

(Davison et al. 2021a). Growth is being driven by an expanding middle class around the world 

and increased interest in healthy eating and gluten free alternatives to grain (Davison et al. 

2021a). The current U.S. consumption of 0.1 pounds per capita would support 20,000 acres of 

chestnuts on U.S. farms. If the United States reached the European level, that would support 

200,000 acres of chestnuts and result in a $1.2 billion chestnut industry. If U.S. consumers 

replaced a portion of the starch they consume in the form of grains and vegetables with 

chestnuts, this level of consumption would support 40,000,000 acres of chestnuts (Davison et al. 

2021a). As noted above, expansion of chestnut planting for food and feed is not likely to use 

Darling 58 American chestnut. Therefore, Darling 58 American chestnuts are anticipated to have 

minimal impacts on trade.

4.8 Compliance with Federal and State Laws and Regulations, Executive Orders, 

Policies, and Treaties 

4.8.1 Federal Laws and Regulations

The laws most relevant to APHIS determinations of regulatory status are the National 

Environmental Policy Act of 1969 (NEPA), the Clean Water Act of 1972 (CWA), the Safe 

Drinking Water Act of 1974 (SDWA), the Clean Air Act of 1970 (CAA), the Endangered 

Species Act of 1973 (ESA), and the National Historic Preservation Act of 1966 (NHPA). 

Compliance with the requirements of the ESA has been addressed in Chapter 6. Compliance with 

the requirements of NEPA, CWA, SDWA, CAA, and NHPA, are specifically addressed in the 

following subsections. 

4.8.2 National Environmental Policy Act (NEPA)

NEPA (42 United States Code (U.S.C) 4321, et seq.) is designed to ensure transparency and 

communication of the possible environmental effects of federal actions prior to implementation. 

The Act and implementing regulations require federal agencies to document, in advance and in 

detail, the potential effects of their actions on the human environment, so as to ensure that there 

is a full understanding of the possible environmental outcomes of federal actions by both the 

decision-makers and the public. This EIS documents the potential environmental outcomes of the 
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alternatives considered, approval or denial of ESF’s petition, and is consistent with the 

requirements of NEPA and Council on Environmental Quality implementing regulations at 40 

CFR parts 1500-1508.

4.8.3 Compliance with the Clean Air Act, Clean Water Act, and Safe Drinking Water Act

The CAA, CWA, and SDWA authorize the EPA to regulate air and water quality in the United 

States. Apart from the blight-tolerance, Darling 58 American chestnut are equivalent to native

chestnut varieties. Therefore, the potential impacts on water resources and air quality are the 

same under both the No Action and Preferred Alternatives. Darling 58 American chestnut are 

expected to be used for forest restoration, which could contribute to potential positive impacts on 

air quality, and potentially water quality. The sources and degree of these impacts would be 

small or no different than that which occurs in current forest systems. As discussed in this EIS, 

the blight-tolerance in Darling 58 American chestnut presents no known risks to water or air 

quality. Considering these factors, a determination of nonregulated status for Darling 58 

American chestnut would not lead to circumstances that resulted in non-compliance with the 

requirements of the CWA, CAA, and SDWA.

4.8.4 Executive Orders Related to Domestic Issues

The following executive orders require consideration of the potential impacts of the Federal 

action to various segments of the population.

• EO 12898 – Federal Actions to Address Environmental Justice in Minority Populations and 

Low-Income Populations 

This EO requires federal agencies to conduct their programs, policies, and activities that 

substantially affect human health or the environment in a manner so as not to exclude 

persons and populations from participation in or benefiting from such programs. It also 

enforces existing statutes to prevent minority and low-income communities from being 

subjected to disproportionately high and adverse human health or environmental effects.

• EO 13045 – Protection of Children from Environmental Health Risks and Safety Risks 

Children may suffer disproportionately from environmental health and safety risks due to 

their developmental stage, higher metabolic rates, and behavior patterns, as compared to 

adults. This EO requires each federal agency to identify, assess, and address the potential 

environmental health and safety risks that may disproportionately affect children.

The No Action and Preferred Alternatives were analyzed with respect to EO 12898 and EO 

13045. Neither alternative evaluated in this EIS is expected to have disproportionate adverse 

impacts on minorities, low-income populations, or children. As reviewed in the EIS, it is highly 

improbable the trait genes and gene products in Darling 58 American chestnut present any risks 

to human health, nor to animal health and welfare. Darling 58 American chestnut would

primarily be used for forest restoration projects and not as a nursery or orchard crop and is 

expected to have overall beneficial impacts on the environment. 

• EO 13175 – Consultation and Coordination with Indian Tribal Governments
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Executive departments and agencies are charged with engaging in consultation and 

collaboration with tribal governments; strengthening the government-to-government 

relationship between the United States and Indian tribes; and reducing the imposition of 

unfunded mandates upon Indian tribes. This EO emphasizes and pledges that federal 

agencies will communicate and collaborate with tribal officials when proposed federal 

actions have potential tribal implications.

Tribal entities are recognized as independent governments and agricultural activities on tribal 

lands would only be conducted if approved by the tribe. Tribes would have control over any 

potential conflict with cultural resources on tribal properties. Approval nor denial of the petition 

is not expected to have any effect on Indian tribal self-governance or sovereignty, tribal treaties, 

or other rights.

Consistent with EO 13175, APHIS sent a letter of notification and request for comment and 

consultation on the proposed action to federally recognized tribes on August 24, 2020. This letter 

contained information regarding the ESF petition request and Darling 58 American chestnut. 

Additionally, this notification asked tribal leaders to contact APHIS if they believed that there 

were potentially significant impacts to tribal lands or resources that should be considered. APHIS 

received two replies, one from the San Manuel Band of Mission Indians, stating they have 

elected to not consult on this announcement, and one from the Ysleta del Sur Pueblo tribe, 

stating they had no comments for APHIS on ESF’s petition request. Additionally, APHIS sent a 

letter informing tribes of APHIS’ intent to prepare an EIS on August 6, 2021. APHIS received 

two replies. One from the Sac & Fox Tribe of the Mississippi in Iowa (Meskwaki Nation) and 

one from the Klamath Tribes, both stating they had no comments for APHIS on ESF’s petition 

request.

APHIS will continue to consult and collaborate with tribal officials to ensure that they are wellinformed and represented in policy and program decisions that may impact their agricultural 

interests, in accordance with EO 13175. A determination of nonregulated status for Darling 58 

American chestnut will not adversely impact cultural resources on tribal properties. 

The No Action and Preferred Alternatives were analyzed with respect to EO 12898, EO 13045, 

and EO 13175. Neither alternative is expected to have a disproportionate adverse impact on 

minorities, low-income populations, or children. Nor is any alternative expected to have potential 

Tribal implications.

• EO 13751 – Safeguarding the Nation from the Impacts of Invasive Species

Invasive species are a significant issue in the United States, causing both adverse economic 

and environmental impacts. This EO directs federal agencies to take action to prevent the 

introduction of invasive species, to provide for their control, and to minimize the economic, 

ecological, and human health impacts that invasive species cause.

American chestnut is not listed in the United States as a noxious weed species by the Federal 

government (USDA-NRCS 2020), nor is it listed as an invasive species by major invasive plant 

data bases. Based on observations and data submitted by the applicant and reviewed by APHIS, 

Darling 58 American chestnut is not expected to become weedy or invasive. While Darling 58 

American chestnut is expected to be resistant to chestnut blight and therefore able to survive 
4-40

unlike other chestnut varieties currently found in Eastern forests, it’s slow spread without human 

intervention, slow natural colonization rates and frequent animal and pest pressure on seeds and 

seedlings (Clark et al. 2014) make it unlikely to be invasive. As part of its PPRA, APHIS 

evaluated the potential weediness and invasiveness of Darling 58 American chestnut and 

concluded that it is unlikely that Darling 58 American chestnut will become weedy or invasive in 

areas where it is grown (USDA-APHIS 2020).

• EO 13186 – Responsibilities of Federal Agencies to Protect Migratory Birds

Federal agencies taking actions that have, or are likely to have, a measurable negative 

effect on migratory bird populations are directed to develop and implement, within two 

years, a Memorandum of Understanding (MOU) with the Fish and Wildlife Service that 

shall promote the conservation of migratory bird populations.

Chestnuts can provide food sources for migratory birds, along migratory routes in North 

America. Migratory birds may visit chestnuts to feed on chestnuts or insects on chestnut trees, 

which provides a valuable source of nutrition to migratory birds. As reviewed in this EIS, it is 

highly unlikely the trait genes and their products present any risks to the health of migratory 

birds. Oxalate oxidase is a common enzyme found in all grains, several other crops and food 

products, and many wild plants, on which birds may forage and the nptII gene and associated 

NPTII enzyme are present in soil and aquatic bacteria and animal gastrointestinal flora (ESF 

2019). The genes and gene products inserted into Darling 58 American chestnut are already 

found in the environment. Because migratory birds that forage on Darling 58 American chestnut 

are unlikely to be adversely affected by ingesting the chestnuts or other plant parts, it is unlikely 

that a determination of nonregulated status for Darling 58 American chestnut would have a 

negative impact on migratory bird populations.

4.9 Conclusions: Potential Impacts on the Human Environment

As discussed in the Scope of Analysis for this EIS (Section 4.1), in considering whether the 

effects of the proposed action could be significant, APHIS analyzed the affected environment 

and degree of the potential effects identified (40 CFR § 1501.3). As part of this analysis APHIS 

considered those requirements outlined in sections 102(2)(C)(ii),(iv), and (v) of NEPA, 40 CFR 

§ 1502.16– Environmental consequences, 40 CFR § 1501.3–Determine the appropriate level of 

NEPA review, 40 CFR § 1502.24–Environmental review and consultation requirements, and 40 

CFR § 1502.15–Affected environment, which are addressed below. 

4.9.1 Adverse environmental effects that cannot be avoided should the proposal be 

implemented.

Planting long lived trees for restoration purposes or for any other purpose is likely to have some 

degree of impact on the environment, as discussed in this EIS. If blight-tolerant American 

chestnut were to become established as an important canopy tree, it would begin to influence 

ecosystem structure and function in these areas, as it did prior to the blight. Impacts to air

quality, surface water and groundwater, soils, biodiversity, and habitats are all possible from 

planting Darling 58 American chestnut. The degree of environmental impacts will depend on a 

variety of factors that include the geographic locale, local biota, weather, inherent soil 

characteristics, and prevalence and diversity of insect pests. Given the historically slow spread 
4-41

rates, low propagule pressure, and need for disturbance to provide sufficient light for fast growth, 

the rate of increase would likely be very slow. Without aggressive restoration efforts, requiring 

considerable effort and coordination at landscape scales, it may require centuries before 

American chestnut becomes a significant presence in the landscape (Gustafson et al. 2017; 

Gustafson et al. 2018). The scale of potential impacts is anticipated to remain relatively minimal 

across the landscape.

4.9.2 The relationship between short-term uses of man’s environment and the maintenance 

and enhancement of long-term productivity. 

Planting of Darling 58 American chestnut would not be considered a short-term use of the 

environment since the trees can live hundreds of years. Long-term productivity depends on the 

sustainable use of natural resources. Restoration with Darling 58 American chestnut is expected 

to occur on long-term research plots and relatively small-scale public horticultural lands before 

being planted on a larger scale in eastern forests and utilize similar resources as other forest 

trees.

4.9.3 Irreversible or irretrievable commitments of resources that would be involved in the 

proposal should it be implemented.

An irreversible or irretrievable commitment of resources refers to impacts on or losses of 

resources that cannot be recovered or reversed. Irreversible commitments of resources are those 

that cause either direct or indirect use of natural resources such that the resources cannot be 

restored or returned to their original condition. Irreversible impacts entail the loss of future 

options, and applies primarily to the use of nonrenewable resources such as fossil fuels, and 

resources that are renewable only over long time spans. Irretrievable is a term that involves the 

loss of productive value or use of resources. For example, certain opportunities can be foregone 

during the conduct of a proposed action, during which a resource cannot be used. These actions 

may be reversible or temporary, but the utilization opportunities foregone during the action are 

irretrievable. 

Production of Darling 58 American chestnut may involve the irreversible consumption of 

nonrenewable resources. The forests of today differ greatly from the forests of a hundred years 

ago in land use, species composition (including many nonnative species not formerly present), 

fire regimes, and climate change. If blight-tolerant American chestnut were to become 

established as an important canopy tree, it would begin to influence ecosystem structure and 

function in these areas, as it did prior to the blight. With an increase in American chestnut, the 

populations of some co-occurring tree species would gradually decline. Chestnut will likely 

replace other tree species in proportion to their abundance, rather than replacing a single species 

or genus. Darling 58 American chestnut spread entails the irretrievable removal of some natural 

habitat as it replaces other tree species from the landscape. 

Renewable and nonrenewable resources utilized for Darling 58 American chestnut production 

would differ little from that of other forest trees. Any irreversible or irretrievable commitments 

of resources in Darling 58 American chestnut production would be the same as or very similar to 

that of other forest trees. Darling 58 American chestnut is expected to be produced on long-term 

research plots and relatively small-scale public horticultural lands before being planted on a 
4-42

larger scale in eastern forests. Restoration efforts will be primarily managed by The American 

Chestnut Foundation. 

4.9.4 Whether the action would violate or conflict with a federal or state laws or local 

requirements governing protection of the environment. 

As reviewed in Section 4.8.3, approval of the petition would not lead to circumstances that 

resulted in non-compliance with any federal, state, or local laws and regulations providing 

protections for environmental and human health. ESF plans to consult with the FDA on the 

safety of Darling 58 American chestnut pursuant to the voluntary consultation process for crop

plants developed using genetic engineering (21 C.F.R. Parts 192 and 592). A food and feed

safety and nutritional assessment of Darling 58 will be submitted to the FDA for review (ESF 

2019).

4.9.5 Possible conflicts between the proposed action and the objectives of federal, regional, 

state, tribal, and local land use plans, policies, and controls for the area concerned. 

There are no conflicts with approval of the petition, and subsequent planting of Darling 58 

American chestnut, with federal, state, tribal, or local land use plans or policies. 

Federal Lands

There are four major federal land management agencies that administer 606.5 million acres (as 

of September 30, 2018). These are the Bureau of Land Management, Fish and Wildlife Service 

(FWS), National Park Service in the Department of the Interior, and the Forest Service in the 

USDA. A fifth agency, the Department of Defense, administers 8.8 million acres in the United 

States (as of September 30, 2017). Together, the five agencies manage about 615.3 million acres, 

or 27% of the U.S. land base (CRS 2020). Many other agencies administer the remaining federal 

acreage. The lands administered by the four major agencies are managed primarily for purposes 

related to preservation, recreation, and development of natural resources (CRS 2020).

APHIS approval of the petition would have no effect on lands governed by federal land 

management agencies. Any planting of Darling 58 American chestnut on federal lands would 

require approval by a federal land management agency. 

Tribal Nations

Executive Order 13175–Consultation and Coordination with Indian Tribal Governments, charges 

executive departments and agencies with engaging in consultation and collaboration with tribal 

governments; strengthening the government-to-government relationship between the United 

States and Indian tribes; and reducing the imposition of unfunded mandates upon Indian tribes. 

The EO emphasizes and pledges that federal agencies will communicate and collaborate with 

tribal officials when proposed federal actions have potential tribal implications. Tribal entities 

are recognized as independent governments and agricultural activities on tribal lands would only 

be conducted if approved by the tribe. Tribes would have control over any potential conflict with 

cultural resources on tribal properties. Approval nor denial of the petition would not have any 

effect on Indian tribal self-governance or sovereignty, tribal treaties, or other rights. APHIS 

conducted outreach to tribal nations informing tribes of ESF’s petition by sending a letter on 

August 24, 2020. APHIS received two replies, one from the San Manuel Band of Mission 
4-43

Indians, stating they have elected to not consult on this announcement, and one from the Ysleta 

del Sur Pueblo tribe, stating they had no comments for APHIS on ESF’s petition request. 

Additionally, APHIS sent a letter informing tribes of APHIS’ intent to prepare an EIS on August 

6, 2021. APHIS received two replies. One from the Sac & Fox Tribe of the Mississippi in Iowa

(Meskwaki Nation) and one from the Klamath Tribes, both stating they had no comments for 

APHIS on ESF’s petition request.

State and Local Land Use Plans and Policies 

The PPA contains a preemption clause (7 U.S.C. § 7756) that prohibits state regulation of any, 

“plant, biological control organism, plant pest, noxious weed, or plant product” to protect against 

plant pests or noxious weeds if the Secretary (USDA) has issued regulations to prevent the 

dissemination of biological control organisms, plant pests, or noxious weeds within the United 

States. The PPA preemption clause does however allow states to impose additional prohibitions 

or restrictions based on special needs supported by sound scientific data or risk assessment. 

Consequently, while the PPA limits states' issuance of laws and regulations governing organisms 

developed using genetic engineering and bars conflicting state regulation, it does allow state 

oversight when there is a special need for additional prohibitions or restrictions.

States use a variety of mechanisms to regulate the movement or release of organisms developed 

using genetic engineering within their jurisdiction. For example, South Dakota simply authorizes 

holders of a federal permit issued under 7 CFR part 340 to use it within the state (SD Stat § 38-

12A-31 (2015)). Minnesota issues state permits for release of organisms developed using genetic 

engineering only after federal applications or permits are on file (MN Stat § 18F.07 (2015)). 

Nebraska may rely on APHIS or other experts before they issue their permit (NE Code § 2-

10,113 (2015)). These illustrative examples show the range of state approaches to regulating the 

movement and release of organisms developed using genetic engineering within state 

boundaries.

Neither of the alternatives considered would affect APHIS partnerships with states in the 

oversight of organisms developed using genetic engineering, to include the planting of Darling 

58 American chestnut. Under both alternatives, APHIS would continue working with states. The 

range of state legislation addressing agricultural biotechnology, namely in the way of permitting, 

crop protection, seed regulation, and economic development, would be unaffected by denial or 

approval of the petition.

4.9.6 Energy requirements and conservation potential of various alternatives and mitigation 

measures. 

The energy requirements involved with the full life cycle of Darling 58 American chestnut

production would differ little from that of other forest trees. However, the conservation potential 

of planting Darling 58 American chestnut could be substantial. American chestnut was 

considered a keystone species, described as "the most important wildlife plant in the eastern 

United States" (Davis 2005), and played a critical role in the function of the overall ecosystem. If 

Darling 58 American chestnuts were to become established, it would influence ecosystem 

structure and function in areas where they were introduced, as American chestnuts did prior to 

the blight (Paillet 2002). Additionally, because American chestnut is fast-growing, long-lived, 

and resistant to decay (Ellison et al. 2005; de Bruijn et al. 2014), its reintroduction could result in 
4-44

increased carbon sequestration and storage in the form of living and dead trees and durable wood 

products (Gustafson et al. 2017). However, given the historically slow spread rates, low 

propagule pressure, and need for disturbance to provide enough light for fast growth, the rate of 

increase would likely be very slow. Without aggressive restoration efforts, requiring 

considerable effort and coordination at landscape scales, it may require centuries before 

conservation impacts are seen (Gustafson et al. 2017; Gustafson et al. 2018).

4.9.7 Natural or depletable resource requirements and conservation potential of various 

alternatives and mitigation measures. 

There are no depletable resource requirements unique to the production of Darling 58 American 

chestnut. Use of natural resources (e.g., irrigation water, soils, fertilizers) would be no different 

than that of other trees in eastern forests. Available mitigation measures to curtail potential 

environmental impacts, such as those summarized below in 4.9.9, would likewise not differ. 

4.9.8 Urban quality, historic and cultural resources, and the design of the built environment, 

including the reuse and conservation potential of various alternatives and mitigation 

measures. 

Darling 58 American chestnut may be grown in proximity to historic or cultural resources. The 

National Historic Preservation Act of 1966 and its implementing regulations (36 CFR part 800) 

requires federal agencies to: 1) determine whether activities they propose constitute 

"undertakings" that have the potential to cause effects on historic properties and 2) if so, to 

evaluate the effects of such undertakings on such historic resources and consult with the 

Advisory Council on Historic Preservation (i.e., State Historic Preservation Office, Tribal 

Historic Preservation Officers), as appropriate. 

Approval of the petition is not a decision that would directly or indirectly result in alteration of 

the character or use of historic properties protected under the NHPA, nor would it result in any 

loss or destruction of cultural or historical resources. There are no proposed major ground 

disturbances; no new physical destruction or damage to property; no alterations of property, 

wildlife habitat, or landscapes; and no prescribed sale, lease, or transfer of ownership of any 

property. This action is limited to a determination of nonregulated status of Darling 58 American 

chestnut. This action would not convert land use to nonagricultural use and, therefore, would 

have no adverse impact on prime farmland. 

Based on these findings, including the assumption that EPA label use instructions are in place to 

protect unique geographic areas and that those label use instructions are adhered to, a 

determination of nonregulated status of Darling 58 American chestnut is not expected to impact 

unique characteristics of geographic areas such as park lands, prime farm lands, wetlands, wild 

and scenic areas, or ecologically critical areas.

4.9.9 Means to mitigate adverse environmental impacts.

Darling 58 American chestnut was developed with the intent of restoring a native tree to its 

former range. The American chestnut was once one of the most abundant trees within its range in 

the eastern United States. As a former keystone species, Darling 58 American chestnut has the 

potential to influence ecosystem structure and function in eastern forests, as it did prior to the 
4-45

blight, if it were to become established. As discussed throughout this EIS, the impacts of a 

determination of nonregulated status for Darling 58 American chestnut are unlikely to be 

adverse. Additionally, because of the historically slow spread rates, low propagule pressure, and 

need for disturbance to provide enough light for fast growth, the rate of increase would likely be 

very slow. Without aggressive restoration efforts, requiring considerable effort and coordination 

at landscape scales, it may require centuries before conservation impacts are seen (Gustafson et 

al. 2017; Gustafson et al. 2018).

4.9.10 Economic and technical considerations, including the economic benefits of the proposed 

action. 

Economic considerations have been evaluated in Section 4.7 Socioeconomics. The economic 

impacts associated with the introduction of Darling 58 American chestnut for restoration would 

be potentially beneficial. 

4.9.11 The degree to which the action may adversely affect the endangered or threatened 

species or its habitat that has been determined to be critical under the Endangered 

Species Act of 1973.

The evaluation of impacts from a determination of nonregulated status of Darling 58 American 

chestnut, and subsequent introduction into the environment, can be found in Appendix 1 of this 

EIS. Darling 58 American chestnut has been found to have no adverse effect on listed species or 

species proposed for listing, and would not affect designated habitat or habitat proposed for 

designation.

4.9.12 The degree to which the proposed action affects public health or safety.

Approval of the petition and subsequent planting of Darling 58 American chestnut would not 

present any risks to public health or worker safety. As reviewed in Section 4.5, it is unlikely that 

humans would be negatively impacted by consuming chestnuts from Darling 58 American 

chestnut. Oxalate oxidase and similar enzymes are consumed and handled daily in cereal grains 

and many other foods without any reports of toxicity, and we have found no evidence that it 

should be considered a toxin (ESF 2019). As discussed previously in this EIS, there are no health 

hazards presented by consumption of Darling 58 American chestnuts. As reviewed in Section 

4.9.4, ESF is consulting with the FDA as to the safety of food and feed derived from Darling 58 

American chestnut (ESF 2019).

4.9.13 Whether the affected environment includes reasonably foreseeable environmental 

trends and planned actions in the affected areas.

Approval of the petition would provide for the release of Darling 58 American chestnut into the 

environment, subject to any FDA consultation, and EPA and state requirements. As of October, 

2021, APHIS has not issued a determination of nonregulated status for any chestnut varieties

developed using genetic engineering. However, APHIS has issues determinations of 

nonregulated status for three fruit trees; apple, papaya, and plum. APHIS maintains a publicly 

available list of petitions and determinations of nonregulated status on its website (USDAAPHIS 2021). Seeds for crops developed using genetic engineering were commercially 
4-46

introduced in the United States for major field crops in 1996, with adoption rates increasing 

rapidly in the years that followed. 

Farmers generally adopt a crop developed using genetic engineering based on the benefits they 

can derive from it, such as effective insect pest or weed control, increased crop yields per acre, 

increased farm net returns, and time savings (Fernandez-Cornejo et al. 2014; Livingston et al. 

2015). Potential net benefits are a function of the particular crop farmed and geographic location; 

agronomic input and market commodity prices; existing on-farm crop production systems; and 

farmer abilities and preferences (Fernandez-Cornejo et al. 2014; Livingston et al. 2015).

Unlike field crops, Darling 58 American chestnut was developed with the intent of restoring a 

native tree to its former range. The American chestnut was once one of the most abundant trees 

within its range in the eastern United States. Because of chestnut blight, the abundance of 

American chestnut drastically declined and was reduced from a dominant overstory tree to a 

small understory shrub (Elliott and Swank 2008; Dalgleish et al. 2015b). As a former keystone 

species, Darling 58 American chestnut has the potential to influence ecosystem structure and 

function in eastern forests, as it did prior to the blight, if it were to become established. As 

discussed throughout this EIS, the impacts of a determination of nonregulated status for Darling 

58 American chestnut are likely to be minimal and require centuries before American chestnut 

becomes a significant enough presence in the landscape to see these impacts (Gustafson et al. 

2017; Gustafson et al. 2018).
4-47

Appendix 1 Threatened and Endangered Species

The Endangered Species Act (ESA) of 1973, as amended, is one of the most far-reaching 

wildlife conservation laws ever enacted by any nation. Congress passed the ESA to prevent 

extinctions facing many species of fish, wildlife and plants. The purpose of the ESA is to 

conserve endangered and threatened species, and the ecosystems on which they depend, as key 

components of America’s heritage. To implement the ESA, the U.S. Fish & Wildlife Service 

(USFWS) works in cooperation with the National Marine Fisheries Service (NMFS), other 

Federal, State, and local agencies, Tribes, non-governmental organizations, and private citizens. 

Before a plant or animal species can receive the protection provided by the ESA, one of the 

Services must first add it to the Federal list of threatened and endangered wildlife and plants.

Threatened and endangered (T&E) species are plants and animals at risk of becoming extinct 

throughout all or part of their geographic range (endangered species) or species likely to become 

endangered in the foreseeable future throughout all or a significant portion of their ranges 

(threatened species).

The Services add a species to the list when they determine it is endangered or threatened because 

of any of the following factors or a combination thereof:

• The present or threatened destruction, modification, or curtailment of its habitat or range;

• Overutilization for commercial, recreational, scientific, or educational purposes;

• Disease or predation;

• The inadequacy of existing regulatory mechanisms; or

• The natural or manmade factors affecting its survival.

Once an animal or plant is added to the list, in accordance with the ESA, protective measures 

apply to the species and its habitat. These measures include protection from adverse effects of 

Federal activities.

Requirements for Federal Agencies

Section 7 (a)(2) of the ESA requires that federal agencies, in consultation with USFWS and/or 

the NMFS, ensure that any action they authorize, fund, or carry out is “not likely to jeopardize 

the continued existence of a listed species or result in the destruction or adverse modification of 

designated critical habitat.” It is the responsibility of the federal agency taking the action to 

assess the effects of their action and to consult with the USFWS and NMFS if it is determined 

that the action “may affect” listed species or designated critical habitat. To facilitate their ESA 

consultation requirements, APHIS met with the USFWS from 1999 to 2003 to discuss factors 

relevant to APHIS’ regulatory authority and effects analysis for petitions for nonregulated status 

and developed a process for conducting an effects determination consistent with the PPA (Title 

IV of Public Law 106-224). APHIS uses this process to help fulfill its obligations and 

responsibilities under Section 7 of the ESA for biotechnology regulatory actions. 

The APHIS regulatory authority over organisms developed using genetic engineering is limited 

to those organisms for which it has reason to believe might be a plant pest or those for which 

APHIS does not have sufficient information to determine that the organism is unlikely to pose a 

plant pest risk. In this case, ESF requests that the USDA APHIS consider that Darling 58 
4-48

American chestnut does not pose a plant pest risk. After completing a Plant Pest Risk 

Assessment, if APHIS determines that Darling 58 American chestnut seeds, plants, or parts 

thereof do not pose a plant pest risk, then this organism would no longer be subject to the plant 

pest provisions of the PPA or to the regulatory requirements of 7 CFR part 340, and therefore, 

APHIS must reach a determination that this organism is no longer regulated. 

As part of its EIS analysis, APHIS analyzed the potential effects of Darling 58 American 

chestnut on the environment, including any potential effects to T&E species and critical habitat. 

As part of this process, APHIS thoroughly reviews product information and data related to the 

organism developed using genetic engineering to inform the ESA effects analysis and, if 

necessary, the biological assessment. For each transgene/transgenic plant the following 

information, data, and questions are considered by APHIS: 

• A review of the biology, taxonomy, and weediness potential of the crop plant and its 

sexually compatible relatives;

• Characterization of each transgene with respect to its structure and function and the 

nature of the organism from which it was obtained;

• A determination of where the new transgene and its products (if any) are produced in the 

plant and their quantity;

• A review of the agronomic performance of the plant including disease and pest 

susceptibilities, weediness potential, and agronomic and environmental impact;

• Determination of the concentrations of known plant toxicants (if any are known in the 

plant);

• Analysis to determine if the transgenic plant is sexually compatible with any T&E 

species of plants or a host of any T&E species; and

• Any other information that may inform the potential for an organism to pose a plant pest

risk.

APHIS met with USFWS officials on June 15, 2011, to discuss and clarify whether APHIS has 

any obligations under the ESA regarding analyzing the effects on T&E species that may occur 

from use of pesticides associated with plants developed using genetic engineering. As a result of 

these joint discussions, USFWS and APHIS have agreed that it is not necessary for APHIS to 

perform an ESA effects analysis on pesticide use associated with plants developed using genetic 

engineering because EPA has both regulatory authority over the labeling of pesticides under 

FIFRA, and the necessary technical expertise to assess pesticide effects on the environment. 

APHIS has no statutory authority to authorize or regulate the use of pesticides by growers. Under 

APHIS’ Part 340 regulations, APHIS only has the authority to regulate Darling 58 American 

chestnut or any organism developed using genetic engineering as long as APHIS believes they 

may pose a plant pest risk. APHIS has no regulatory jurisdiction over any other risks associated 

with organisms developed using genetic engineering including risks resulting from the use of 

pesticides with those organisms.

In following this review process, APHIS, as described below, has evaluated the potential effects 

that a determination of nonregulated status of Darling 58 American chestnut may have, if any, on 

federally-listed T&E species (mammals, bird, amphibians, reptiles, invertebrates, fish and 

shellfish) and species proposed for listing, as well as designated critical habitat and habitat 
4-49

proposed for designation. APHIS also considered potential effects of Darling 58 planted outside 

the historic range of the American chestnut. While planting outside the range of conventional 

American chestnut is not part of this petition, APHIS considered it reasonably certain to occur in 

the event Darling 58 American chestnut is made available to the public. However, American 

chestnut trees spread at an average rate of “no more than a few kilometers per century” (Paillet 

and Rutter 1989) and will likely take centuries for chestnut to have a significant presence on the 

landscape. Considering this, and since planting outside the historic range would likely occur for 

non-restoration purposes in anthropogenically modified environments such as landscaping, 

parks, and arboretums, APHIS does not expect Darling 58 American chestnut to have negative 

effects on T&E species outside its historic range (Figure 1). 

Potential Effects of Darling 58 American Chestnut on T&E Species

While a determination of nonregulated status for Darling 58 American chestnut would allow for 

plantings anywhere within the United States, APHIS limited the action area to the 25 states 

where American chestnut historically occurred (Figure 1). Darling 58 American chestnut is 

intended to be used as a restoration tree to establish and colonize much of the eastern United 

States where stunted trees persist as stumps and small trees. APHIS obtained and reviewed the 

USFWS list of T&E species for these states from the USFWS Environmental Conservation 

Online System (USFWS 2022). The species list can be found at the end of this appendix. Our 

analysis focused on potential effects on terrestrial, avian, and aquatic species that could come in 

contact with and otherwise carry out life functions (nesting, feeding, rearing, etc) in the vicinity 

or on Darling 58 American chestnut tree itself within the action area, where American chestnut 

was once the dominant forest tree (Figure 1). Accordingly, the issues discussed herein focus on 

the potential environmental consequences of approval of the petition for nonregulated status of 

Darling 58 American chestnut on T&E species and critical habitat in the action area, where 

Darling 58 could be planted.

For its analysis on T&E plants and critical habitat, APHIS focused on the potential differences 

between Darling 58 American chestnut and conventional American chestnut; the potential for 

ecosystem effects; and the potential for gene movement to native plants, listed species, and 

species proposed for listing. According to information submitted by the petitioner and reviewed 

by APHIS, Darling 58 American chestnut is phenotypically and biochemically comparable to 

conventional American Chestnut (ESF 2019). No significant differences have been observed in 

terms of plant pest risk traits such as competitiveness, responses to other pests, interactions with 

other organisms in the environment, or survival (besides blight tolerance) (ESF 2019). Therefore, 

Darling 58 American chestnut should present no additional weediness traits or plant pest risks 

than wild-type American chestnuts or traditionally bred hybrids. The American chestnut is not 

considered an invasive, fast colonizing tree, and the OxO gene will not change these traits.

Therefore, Darling 58 American chestnut is not expected to invade or alter critical habitat in 

ways that would be detrimental to T&E species. Areas that are not intentionally planted with 

blight-tolerant chestnuts will likely remain without chestnuts for decades or longer (ESF 2019).

Darling 58 American chestnut could effectively be excluded from critical habitat if needed.

Independent nutrition analyses have confirmed that transgenic chestnuts are not nutritionally 

different than their wild-type relatives. Even with the ubiquity of OxO in the environment and 
4-50

agriculture, there are no reports of this enzyme being detrimental to human or animal health, 

having adverse effects on the environment, or being a plant pest risk.

For its analysis of effects on T&E animals, APHIS focused on the implications of exposure to 

the OxO gene expressed in Darling 58 American chestnut as well as the tree itself, and the ability 

of the Darling 58 American chestnut to serve as a host or forage. For plant species, APHIS 

focused on the implications of displacement and the potential for gene flow to other plant and 

tree species including its potential to hybridize for a T&E Species.

Threatened and Endangered Plant Species and Critical Habitat

Data provided by ESF (2019) were used in the APHIS analysis of the weediness potential for 

Darling 58 American chestnut, and further evaluated for the potential to impact T&E species and 

critical habitat.

No differences were detected between Darling 58 chestnut and conventional chestnut in growth, 

reproduction, or interactions with pests and diseases, other than the intended effect of resistance

to chestnut blight (ESF 2019). American chestnut trees spread at an average rate of “no more 

than a few kilometers per century” (Paillet and Rutter 1989). Further, slow natural colonization 

rates and frequent animal and pest pressure on seeds and seedlings (Clark et al. 2014) suggest 

chestnuts, regardless of transgene status, will not rapidly invade new areas (Cook and Forest 

1978). Accordingly, APHIS has concluded that the determination of nonregulated status of 

Darling 58 American chestnut does not present a plant pest risk, does not present an increased

risk of weediness, and does not present an increased risk of gene flow.

Chestnuts were replaced mainly by oak and maple, and to a lesser extent, hickory, birch, black 

cherry, and others as dominant canopy tree species (Keever 1953; Stephenson 1986; Stephenson 

et al. 1991; Brewer 1995). No single species has emerged in a dominant role across a broad 

geographic range, comparable to the pre-blight status of American chestnut. With an increase in 

American chestnut (Darling 58), some co-occurring tree species would gradually decline, most 

likely the same trees that replaced American chestnut after the blight was introduced. As 

discussed above, American chestnut would likely replace other tree species in proportion to their 

abundance, rather than replacing a single species or genus (Gustafson et al. 2017). Given the 

relatively close overlap between the niches of chestnut and oak (Keever 1953) competition from 

chestnut would likely affect oaks more than other species (Gustafson et al. 2017). There is no 

reason to believe that the reintroduction of American chestnut would result in reduction of any 

competing tree species to threatened or endangered levels or reduce any tree species to such a 

level as to impact T&E species depending on those resources. Darling 58 American chestnut is 

not expected to invade or alter critical habitat in ways that would be detrimental to T&E species.

Areas that are not intentionally planted with blight-tolerant chestnuts will likely remain without 

chestnuts for decades or longer (ESF 2019). Darling 58 American chestnut could effectively be 

excluded from critical habitat if needed.

Studies conducted by the petitioner showed there were no significant differences in colonization 

by ectomycorrhizal fungi in roots compared to non-transgenic controls, suggesting that the 

presence or expression of OxO in Darling 58 American chestnut does not pose risks to native soil 
4-51

fungi that are ecologically important for American chestnuts and other trees (ESF 2019). As 

such, Darling 58 American chestnut is not expected to have impacts on soil quality. 

APHIS also evaluated the potential of Darling 58 American chestnut to cross with a listed 

species. None of the relatives of Darling 58 American chestnut are Federally listed (or proposed) 

as endangered or threatened species (http://www.fws.gov/endangered/). Therefore, the presence 

of Darling 58 chestnut in the environment will not result in movement of the inserted genetic 

material to any endangered or threatened species. 

After reviewing the list of threatened and endangered plant species, and laboratory and field data 

that confirms no sexual compatibility of any T&E species with Darling 58 American chestnut, as 

well as studies that confirm no negative effects to soil quality, that Darling 58 American chestnut 

would not reduce competing trees to threatened or endangered levels, and that Darling 58 

American chestnut could be effectively excluded from critical habitat if needed, APHIS 

determined that Darling 58 American chestnut will have no effect on threatened or endangered 

plant species or on their critical habitat.

Threatened and Endangered Animal Species and Critical Habitat

Threatened and endangered animal vertebrate and invertebrate species that may be exposed to 

Darling 58 American chestnut would be those T&E species that inhabit areas where Darling 58 

American chestnut would be planted for restoration purposes. 

APHIS considered the possibility that Darling 58 American chestnut could serve as an ecosystem 

component for a threatened or endangered species (i.e., listed invertebrates or other organisms 

that may use the Darling 58 American chestnut tree to complete its lifecycle). APHIS reviewed 

the complete T&E species database available on the FWS website (USFWS 2022) and found 

several animal groups that would use Darling 58 chestnut as an important component in the 

completion of its lifecycle. 

MAMMALS 

Pre-blight, American chestnut was described as "the most important wildlife plant in the eastern 

United States" (Davis 2005). Native mammals such as white-tailed deer (Odocoileus 

virginianus), cottontail rabbit (Sylvilagus floridanus), and black bears (Ursus americanus) all 

consumed chestnuts (Diamond et al. 2000; Wang et al. 2013), as well as many other vertebrates 

including rodents (Lichti et al. 2014) and birds including wild turkeys (Meleagris gallopavo), 

American crow (Corvus brachyrhynchos), blue jay (Cyanocitta cristata), ruffed grouse (Bonasa 

umbellus), and the extinct passenger pigeon (Ectopistes migratorius) (Webb 1986; Russell 1987) 

and heath hen (Hill 1992). Other mammalian wildlife benefit indirectly from the American 

chestnut. Bats, weasels, mountain lion, bobcat, and other predators prey on rodents and other 

organisms that consume American chestnuts. As discussed below, the decline of the American 

chestnut likely had far-reaching effects on food chains that disrupted the entire forest ecosystem. 

Studies suggest the decline in the Allegheny wood rat (a Federal species of concern) may have 

coincided with the reduction in American chestnut, noting the chestnut is an important food 

source for the rodent (Wright and Kirkland 2000). Other wildlife such as squirrels may have
4-52

declined greatly as a result of habitat destruction and loss of chestnut hard mast during fall and 

winter, and avian and mammalian predators also decreased as a result of decline of these prey 

populations (Hill 1992). An important feature of the American chestnut is its reliability to 

produce mast (flowers and nuts) compared to other nut-producing trees that have out-competed 

the chestnut since the blight. In addition to producing more mast, chestnut was the most reliable 

of all the nut-producing trees in the eastern forest. Because of its relatively late bloom (May 

through June), it could produce mast despite late-spring frost, unlike almost all other mast food 

sources. Many other trees, including oaks and hickories, avoid the effects of granivores (seed and 

grain consumers) eating all their nuts by surprising them with intermittent high-mast years

(Burke 2013). This evidence suggests forest restoration with Darling 58 chestnut may reduce 

systematic impacts on ecosystem food chains as both predatory and prey mammals increase 

survival, benefiting the entire forest ecosystem, including threatened and endangered mammals

INVERTEBRATES

American chestnut provided a food source to numerous insect species, especially during the 

flowering period. The blight and subsequent reduction in American chestnut is thought to have 

caused at least five indigenous insect species to become extinct or extremely rare (Office of 

Technology Assessment 1993). Pollen feeders in the insect orders coleoptera (beetles), 

lepidoptera (moths), diptera (flies), hymenoptera (bees) and others have been observed visiting 

chestnut catkins (Clapper 1954; Opler 1978; Hasegawa et al. 2015; Tumminello 2016; Zirkle 

2017). Darling 58 American chestnut is compositionally equivalent to and as safe and nutritious 

as the forage produced from conventional American chestnut (ESF 2019). Since American 

chestnut provided a more stable and more abundant source of mast than oaks, hickory, and beech 

species (Diamond et al. 2000), if Darling 58 American chestnuts were to become established it 

could result in population increases of the animal and insect species that feed on chestnut (Hill 

1992). The increase would be most pronounced during years of low seed production by other 

masting species, resulting in less fluctuation in those species that consume seeds (Dalgleish and 

Swihart 2012). 

AQUATIC SPECIES

The replacement of oak by Darling 58 American chestnut may also increase macroinvertebrate 

activity with potential consequences on population, community, and ecosystem levels since 

chestnut leaf litter is of higher nutritional value for aquatic invertebrates than oak (Smock and 

MacGregor 1988). Over the long term, this could help restore functional habitat for those T&E 

species that use aquatic invertebrates for forage, primarily fish species. Further, slow decaying 

chestnut wood may increase stream channel complexity as it replaces faster decaying species 

such as oak. Over time, this would provide additional conservation value for critical habitat for 

T&E fish and invertebrates (Ellison et al. 2005).

Introduction of Darling 58 American chestnut should not impact overall water quality resources 

in the action area used by listed animal and insect species. American chestnut does not have 

different water requirements than those tree species currently found in the action area (ESF 

2019).

AVIANS
4-53

Migratory birds may visit chestnuts to feed on chestnuts or insects on chestnut trees, which 

provides a valuable source of nutrition to migratory birds. As reviewed in this EIS, it is highly 

unlikely the trait genes and their products present any risks to the health of migratory birds.

Oxalate oxidase is a common enzyme found in all grains, several other crops and food products, 

and many wild plants, on which birds may forage and the nptII gene and associated NPTII 

enzyme are present in soil and aquatic bacteria and animal gastrointestinal flora (ESF 2019). The 

genes and gene products inserted into Darling 58 American chestnut are already found in the 

environment. Because migratory birds and other avian species that forage on Darling 58 

American chestnut are unlikely to be adversely affected by ingesting the chestnuts or other plant 

parts, it is unlikely that a determination of nonregulated status for Darling 58 American chestnut 

would have any effect on threatened and endangered migratory bird populations.

ESF plans to consult with the FDA on the safety of Darling 58 American chestnut pursuant to the 

voluntary consultation process for crop plants developed using genetic engineering (21 C.F.R. 

Parts 192 and 592). A food and feed safety and nutritional assessment of Darling 58 American 

chestnut will be submitted to the FDA for review (ESF 2019).

Summary of Effects and Determination

As discussed above, several experiments have been performed on OxO-expressing American 

chestnuts, and results consistently confirm a lack of plant pest risks or non-target effects. Studies 

have been conducted on Darling 58 American chestnut, offspring of Darling 58 American 

chestnut, and on older legacy events that also express OxO. These experiments included 

observing mycorrhizal colonization of chestnut roots, aquatic and terrestrial insect herbivory on 

leaves, wood frog tadpoles feeding on leaf litter, leaf litter decomposition, interactions with 

nearby plants, and use by bumble bees of OxO-containing chestnut pollen. Nutritional 

composition and tannin concentrations of the OxO-containing nuts have been evaluated by 

commercial testing labs, and the OxO enzyme was queried against allergen, gluten, and toxin 

databases. In all cases, the blight-tolerant transgenic American chestnut trees were shown to be 

equivalent to wild-type American or traditionally bred hybrid chestnuts (ESF 2019). No effects 

on T&E species are expected from the OxO gene in Darling 58 American chestnut. 

After reviewing the possible effects of determining nonregulated status of Darling 58 American 

chestnut, APHIS has not identified any stressor that could negatively affect the reproduction, 

numbers, or distribution of a listed T&E species or species proposed for listing. APHIS also 

considered the potential effect of a determination of nonregulated status of Darling 58 American 

chestnut on designated critical habitat or habitat proposed for designation and could identify no 

differences from effects that would occur from the current state of eastern forests. Consumption 

of Darling 58 American chestnut nuts, wood or leaf material by any listed species or species 

proposed for listing will not result in a toxic or allergic reaction. While APHIS determined that 

the planting of Darling 58 American chestnut will likely result in some degree of impacts to the 

environment, negative impacts to threatened and endangered species or critical habitat that occur 

where it would be planted is unlikely. Any long-term benefits wouldn’t be measurable for 

decades or longer. As such, no effects are expected to listed and proposed T&E species and 

critical habitat where Darling 58 American chestnut would be planted. 
4-54

Based on these factors, APHIS has concluded that a determination of nonregulated status of 

Darling 58 American chestnut, will have no effect on listed species or species proposed for 

listing, and would not affect designated habitat or habitat proposed for designation. Because of 

this no-effect determination, consultation under Section 7(a)(2) of the Act or the concurrences of 

the USFWS or NMFS is not required. 

Table 2. List of T&E Species within American Chestnut Historic Range

Scientific Name Common name Status Group

Acipenser oxyrinchus 

(=oxyrhynchus) desotoi

Gulf sturgeon Threatened Fishes

Aconitum noveboracense Northern wild monkshood Threatened Flowering Plants

Aeschynomene virginica Sensitive joint-vetch Threatened Flowering Plants

Agalinis acuta Sandplain gerardia Endangered Flowering Plants

Alasmidonta atropurpurea Cumberland elktoe Endangered Clams

Alasmidonta heterodon Dwarf wedgemussel Endangered Clams

Alasmidonta raveneliana Appalachian elktoe Endangered Clams

Alligator mississippiensis American alligator Similarity of 

Appearance 

(Threatened)

Reptiles

Amaranthus pumilus Seabeach amaranth Threatened Flowering Plants

Amblema neislerii Fat threeridge (mussel) Endangered Clams

Ambystoma bishopi Reticulated flatwoods 

salamander

Endangered Amphibians

Ambystoma cingulatum Frosted Flatwoods salamander Threatened Amphibians

Amphianthus pusillus Little amphianthus Threatened Flowering Plants

Anguispira picta Painted snake coiled forest snail Threatened Snails

Antrolana lira Madison Cave isopod Threatened Crustaceans

Apios priceana Prices potato-bean Threatened Flowering Plants

Arabis georgiana Georgia rockcress Threatened Flowering Plants

Arabis perstellata Braun's rock-cress Endangered Flowering Plants

Asclepias meadii Mead's milkweed Threatened Flowering Plants

Asplenium scolopendrium 

var. americanum

American hart's-tongue fern Threatened Ferns and Allies

Astragalus bibullatus Guthrie's (=Pyne's) ground-plum Endangered Flowering Plants

Astragalus robbinsii var. 

jesupii

Jesup''s milk-vetch Endangered Flowering Plants

Athearnia anthonyi Anthony's riversnail Endangered Snails

Baptisia arachnifera Hairy rattleweed Endangered Flowering Plants

Betula uber Virginia round-leaf birch Threatened Flowering Plants

Boechera serotina Shale barren rock cress Endangered Flowering Plants

Bombus affinis Rusty patched bumble bee Endangered Insects

Brychius hungerfordi Hungerford's crawling water 

Beetle

Endangered Insects

Calidris canutus rufa Red knot Threatened Birds

Cambarus callainus Big Sandy crayfish Threatened Crustaceans
4-55

Scientific Name Common name Status Group

Cambarus cracens Slenderclaw crayfish Endangered Crustaceans

Cambarus veteranus Guyandotte River crayfish Endangered Crustaceans

Campeloma decampi Slender campeloma Endangered Snails

Canis lupus Gray wolf Endangered Mammals

Cardamine micranthera Small-anthered bittercress Endangered Flowering Plants

Caretta caretta Loggerhead sea turtle Threatened Reptiles

Carex lutea Golden sedge Endangered Flowering Plants

Charadrius melodus Piping Plover Endangered Birds

Chelonia mydas Green sea turtle Threatened Reptiles

Chrosomus saylori Laurel dace Endangered Fishes

Cirsium pitcheri Pitcher's thistle Threatened Flowering Plants

Clematis morefieldii Morefields leather flower Endangered Flowering Plants

Clematis socialis Alabama leather flower Endangered Flowering Plants

Conradina verticillata Cumberland rosemary Threatened Flowering Plants

Corynorhinus (=Plecotus) 

townsendii virginianus

Virginia big-eared bat Endangered Mammals

Cottus paulus (=pygmaeus) Pygmy Sculpin Threatened Fishes

Crystallaria cincotta diamond Darter Endangered Fishes

Cumberlandia monodonta Spectaclecase (mussel) Endangered Clams

Cyprinella caerulea Blue shiner Threatened Fishes

Cyprogenia stegaria Fanshell Endangered Clams

Dalea foliosa Leafy prairie-clover Endangered Flowering Plants

Dermochelys coriacea Leatherback sea turtle Endangered Reptiles

Dromus dromas Dromedary pearlymussel Endangered Clams

Drymarchon couperi Eastern indigo snake Threatened Reptiles

Echinacea laevigata Smooth coneflower Endangered Flowering Plants

Elassoma alabamae Spring pygmy sunfish Threatened Fishes

Elimia crenatella Lacy elimia (snail) Threatened Snails

Ellipsoptera puritana Puritan tiger beetle Threatened Insects

Elliptio chipolaensis Chipola slabshell Threatened Clams

Elliptio lanceolata Yellow lance Threatened Clams

Elliptio spinosa Altamaha Spinymussel Endangered Clams

Elliptoideus sloatianus Purple bankclimber (mussel) Threatened Clams

Epioblasma brevidens Cumberlandian combshell Endangered Clams

Epioblasma capsaeformis Oyster mussel Endangered Clams

Epioblasma florentina 

florentina

Yellow blossom (pearlymussel) Endangered Clams

Epioblasma florentina 

walkeri (=E. walkeri)

Tan riffleshell Endangered Clams

Epioblasma metastriata Upland combshell Endangered Clams

Epioblasma obliquata 

obliquata

Purple Cat's paw (=Purple Cat's 

paw pearlymussel)

Endangered Clams
4-56

Scientific Name Common name Status Group

Epioblasma obliquata 

perobliqua

White catspaw (pearlymussel) Endangered Clams

Epioblasma othcaloogensis Southern acornshell Endangered Clams

Epioblasma penita Southern combshell Endangered Clams

Epioblasma rangiana Northern riffleshell Endangered Clams

Epioblasma torulosa 

gubernaculum

Green blossom (pearlymussel) Endangered Clams

Epioblasma torulosa 

torulosa

Tubercled blossom 

(pearlymussel)

Endangered Clams

Epioblasma triquetra Snuffbox mussel Endangered Clams

Epioblasma turgidula Turgid blossom (pearlymussel) Endangered Clams

Eretmochelys imbricata Hawksbill sea turtle Endangered Reptiles

Erimonax monachus Spotfin Chub Threatened Fishes

Erimystax cahni Slender chub Threatened Fishes

Etheostoma akatulo bluemask darter Endangered Fishes

Etheostoma boschungi Slackwater darter Threatened Fishes

Etheostoma chermocki Vermilion darter Endangered Fishes

Etheostoma chienense Relict darter Endangered Fishes

Etheostoma etowahae Etowah darter Endangered Fishes

Etheostoma nuchale Watercress darter Endangered Fishes

Etheostoma osburni Candy darter Endangered Fishes

Etheostoma percnurum Duskytail darter Endangered Fishes

Etheostoma phytophilum Rush Darter Endangered Fishes

Etheostoma rubrum Bayou darter Threatened Fishes

Etheostoma scotti Cherokee darter Threatened Fishes

Etheostoma sellare Maryland darter Endangered Fishes

Etheostoma spilotum Kentucky arrow darter Threatened Fishes

Etheostoma susanae Cumberland darter Endangered Fishes

Etheostoma trisella Trispot darter Threatened Fishes

Etheostoma wapiti Boulder darter Endangered Fishes

Fundulus julisia Barrens topminnow Endangered Fishes

Fusconaia burkei Tapered pigtoe Threatened Clams

Fusconaia cor Shiny pigtoe Endangered Clams

Fusconaia cuneolus Finerayed pigtoe Endangered Clams

Fusconaia escambia Narrow pigtoe Threatened Clams

Fusconaia masoni Atlantic pigtoe Threatened Clams

Geum radiatum Spreading avens Endangered Flowering Plants

Glaucomys sabrinus 

coloratus

Carolina northern flying squirrel Endangered Mammals

Glyptemys muhlenbergii bog turtle Similarity of 

Appearance 

(Threatened)

Reptiles

Gopherus polyphemus Gopher tortoise Threatened Reptiles

Graptemys flavimaculata Yellow-blotched map turtle Threatened Reptiles
4-57

Scientific Name Common name Status Group

Graptemys oculifera Ringed map turtle Threatened Reptiles

Grus canadensis pulla Mississippi sandhill crane Endangered Birds

Gymnoderma lineare Rock gnome lichen Endangered Lichens

Habroscelimorpha dorsalis 

dorsalis

Northeastern beach tiger beetle Threatened Insects

Hamiota altilis Finelined pocketbook Threatened Clams

Hamiota australis Southern Sandshell Threatened Clams

Hamiota perovalis Orangenacre mucket Threatened Clams

Hamiota subangulata Shinyrayed pocketbook Endangered Clams

Hedyotis purpurea var. 

montana

Roan Mountain bluet Endangered Flowering Plants

Helenium virginicum Virginia sneezeweed Threatened Flowering Plants

Helianthus schweinitzii Schweinitz's sunflower Endangered Flowering Plants

Helianthus verticillatus Whorled Sunflower Endangered Flowering Plants

Helonias bullata Swamp pink Threatened Flowering Plants

Hemistena lata Cracking pearlymussel Endangered Clams

Hexastylis naniflora Dwarf-flowered heartleaf Threatened Flowering Plants

Hudsonia montana Mountain golden heather Threatened Flowering Plants

Hymenoxys herbacea Lakeside daisy Threatened Flowering Plants

Iliamna corei Peter's Mountain mallow Endangered Flowering Plants

Iris lacustris Dwarf lake iris Threatened Flowering Plants

Isoetes louisianensis Louisiana quillwort Endangered Ferns and Allies

Isoetes melanospora Black spored quillwort Endangered Ferns and Allies

Isoetes tegetiformans Mat-forming quillwort Endangered Ferns and Allies

Isotria medeoloides Small whorled pogonia Threatened Flowering Plants

Lampsilis abrupta Pink mucket (pearlymussel) Endangered Clams

Lampsilis virescens Alabama lampmussel Endangered Clams

Lasmigona decorata Carolina heelsplitter Endangered Clams

Laterallus jamaicensis ssp. 

jamaicensis

Eastern Black rail Threatened Birds

Leavenworthia crassa Fleshy-fruit gladecress Endangered Flowering Plants

Leavenworthia exigua

laciniata

Kentucky glade cress Threatened Flowering Plants

Lemiox rimosus Birdwing pearlymussel Endangered Clams

Lepidochelys kempii Kemp's ridley sea turtle Endangered Reptiles

Leptoxis ampla Round rocksnail Threatened Snails

Leptoxis foremani Interrupted (=Georgia) 

Rocksnail

Endangered Snails

Leptoxis plicata Plicate rocksnail Endangered Snails

Leptoxis taeniata Painted rocksnail Threatened Snails

Lepyrium showalteri Flat pebblesnail Endangered Snails

Lesquerella lyrata Lyrate bladderpod Threatened Flowering Plants

Lesquerella perforata Spring Creek bladderpod Endangered Flowering Plants
4-58

Scientific Name Common name Status Group

Liatris helleri Heller's blazingstar Threatened Flowering Plants

Lindera melissifolia Pondberry Endangered Flowering Plants

Lioplax cyclostomaformis Cylindrical lioplax (snail) Endangered Snails

Lirceus usdagalun Lee County cave isopod Endangered Crustaceans

Lycaeides melissa samuelis Karner blue butterfly Endangered Insects

Lynx canadensis Canada Lynx Threatened Mammals

Lysimachia asperulaefolia Rough-leaved loosestrife Endangered Flowering Plants

Margaritifera marrianae Alabama pearlshell Endangered Clams

Marshallia mohrii Mohr's Barbara's buttons Threatened Flowering Plants

Marstonia ogmorhaphe Royal marstonia (snail) Endangered Snails

Marstonia pachyta Armored snail Endangered Snails

Medionidus acutissimus Alabama moccasinshell Threatened Clams

Medionidus parvulus Coosa moccasinshell Endangered Clams

Medionidus penicillatus Gulf moccasinshell Endangered Clams

Medionidus simpsonianus Ochlockonee moccasinshell Endangered Clams

Medionidus walkeri Suwannee moccasinshell Threatened Clams

Menidia extensa Waccamaw silverside Threatened Fishes

Mesodon clarki nantahala noonday snail Threatened Snails

Microhexura montivaga Spruce-fir moss spider Endangered Arachnids

Mimulus michiganensis Michigan monkey-flower Endangered Flowering Plants

Mycteria americana Wood stork Threatened Birds

Myotis grisescens Gray bat Endangered Mammals

Myotis septentrionalis Northern Long-Eared Bat Threatened Mammals

Myotis sodalis Indiana bat Endangered Mammals

Necturus alabamensis Black warrior (=Sipsey Fork) 

Waterdog

Endangered Amphibians

Necturus lewisi Neuse River waterdog Threatened Amphibians

Neonympha mitchellii 

francisci

Saint Francis' satyr butterfly Endangered Insects

Neonympha mitchellii 

mitchellii

Mitchell's satyr Butterfly Endangered Insects

Nerodia erythrogaster 

neglecta

Copperbelly water snake Threatened Reptiles

Nicrophorus americanus American burying beetle Threatened Insects

Notropis albizonatus Palezone shiner Endangered Fishes

Notropis cahabae Cahaba shiner Endangered Fishes

Notropis mekistocholas Cape Fear shiner Endangered Fishes

Noturus baileyi Smoky madtom Endangered Fishes

Noturus crypticus Chucky Madtom Endangered Fishes

Noturus flavipinnis Yellowfin madtom Threatened Fishes

Noturus furiosus Carolina madtom Endangered Fishes

Noturus stanauli Pygmy madtom Endangered Fishes

Noturus trautmani Scioto madtom Endangered Fishes
4-59

Scientific Name Common name Status Group

Novisuccinea 

chittenangoensis

Chittenango ovate amber snail Threatened Snails

Oarisma poweshiek Poweshiek skipperling Endangered Insects

Obovaria choctawensis Choctaw bean Endangered Clams

Obovaria retusa Ring pink (mussel) Endangered Clams

Orconectes shoupi Nashville crayfish Endangered Crustaceans

Oxypolis canbyi Canby's dropwort Endangered Flowering Plants

Palaemonias alabamae Alabama cave shrimp Endangered Crustaceans

Palaemonias ganteri Kentucky cave shrimp Endangered Crustaceans

Parvaspina collina James spinymussel Endangered Clams

Parvaspina steinstansana Tar River spinymussel Endangered Clams

Pedicularis furbishiae Furbish lousewort Endangered Flowering Plants

Pegias fabula Littlewing pearlymussel Endangered Clams

Percina antesella Amber darter Endangered Fishes

Percina aurolineata Goldline darter Threatened Fishes

Percina aurora Pearl darter Threatened Fishes

Percina jenkinsi Conasauga logperch Endangered Fishes

Percina rex Roanoke logperch Endangered Fishes

Percina tanasi Snail darter Threatened Fishes

Peromyscus polionotus 

ammobates

Alabama beach mouse Endangered Mammals

Phaeognathus hubrichti Red Hills salamander Threatened Amphibians

Phoxinus cumberlandensis Blackside dace Threatened Fishes

Physaria globosa Short's bladderpod Endangered Flowering Plants

Picoides borealis Red-cockaded woodpecker Endangered Birds

Pituophis melanoleucus 

lodingi

Black pinesnake Threatened Reptiles

Pityopsis ruthii Ruth's golden aster Endangered Flowering Plants

Platanthera integrilabia White fringeless orchid Threatened Flowering Plants

Platanthera leucophaea Eastern prairie fringed orchid Threatened Flowering Plants

Plethobasus cicatricosus White wartyback (pearlymussel) Endangered Clams

Plethobasus cooperianus Orangefoot pimpleback 

(pearlymussel)

Endangered Clams

Plethobasus cyphyus Sheepnose Mussel Endangered Clams

Plethodon nettingi Cheat Mountain salamander Threatened Amphibians

Plethodon shenandoah Shenandoah salamander Endangered Amphibians

Pleurobema clava Clubshell Endangered Clams

Pleurobema curtum Black clubshell Endangered Clams

Pleurobema decisum Southern clubshell Endangered Clams

Pleurobema furvum Dark pigtoe Endangered Clams

Pleurobema georgianum Southern pigtoe Endangered Clams

Pleurobema hanleyianum Georgia pigtoe Endangered Clams

Pleurobema marshalli Flat pigtoe Endangered Clams
4-60

Scientific Name Common name Status Group

Pleurobema perovatum Ovate clubshell Endangered Clams

Pleurobema plenum Rough pigtoe Endangered Clams

Pleurobema pyriforme Oval pigtoe Endangered Clams

Pleurobema strodeanum Fuzzy pigtoe Threatened Clams

Pleurobema taitianum Heavy pigtoe Endangered Clams

Pleurocera foremani Rough hornsnail Endangered Snails

Pleuronaia dolabelloides Slabside Pearlymussel Endangered Clams

Pleuronaia gibber Cumberland pigtoe Endangered Clams

Polygyriscus virginianus Virginia fringed mountain snail Endangered Snails

Potamilus capax Fat pocketbook Endangered Clams

Potamilus inflatus Inflated heelsplitter Threatened Clams

Pseudemys alabamensis Alabama red-bellied turtle Endangered Reptiles

Pseudemys rubriventris 

bangsi

Plymouth Redbelly Turtle Endangered Reptiles

Ptilimnium nodosum Harperella Endangered Flowering Plants

Ptychobranchus greenii Triangular Kidneyshell Endangered Clams

Ptychobranchus jonesi Southern kidneyshell Endangered Clams

Ptychobranchus subtentus Fluted kidneyshell Endangered Clams

Quadrula cylindrica 

cylindrica

Rabbitsfoot Threatened Clams

Quadrula cylindrica 

strigillata

Rough rabbitsfoot Endangered Clams

Quadrula fragosa Winged Mapleleaf Endangered Clams

Quadrula stapes Stirrupshell Endangered Clams

Rana sevosa dusky gopher frog Endangered Amphibians

Reginaia rotulata Round Ebonyshell Endangered Clams

Rhodiola integrifolia ssp. 

leedyi

Leedy's roseroot Threatened Flowering Plants

Rhus michauxii Michaux's sumac Endangered Flowering Plants

Rhynchospora knieskernii Knieskern's Beaked-rush Threatened Flowering Plants

Ribes echinellum Miccosukee gooseberry Threatened Flowering Plants

Sagittaria fasciculata Bunched arrowhead Endangered Flowering Plants

Sagittaria secundifolia Kral's water-plantain Threatened Flowering Plants

Salmo salar Atlantic salmon Endangered Fishes

Sarracenia oreophila Green pitcher-plant Endangered Flowering Plants

Sarracenia rubra ssp. 

alabamensis

Alabama canebrake pitcher-plant Endangered Flowering Plants

Sarracenia rubra ssp. 

jonesii

Mountain sweet pitcher-plant Endangered Flowering Plants

Scaphirhynchus albus Pallid sturgeon Endangered Fishes

Scaphirhynchus suttkusi Alabama sturgeon Endangered Fishes

Schwalbea americana American chaffseed Endangered Flowering Plants

Scirpus ancistrochaetus Northeastern bulrush Endangered Flowering Plants

Scutellaria montana Large-flowered skullcap Threatened Flowering Plants
4-61

Scientific Name Common name Status Group

Silene polypetala Fringed campion Endangered Flowering Plants

Sistrurus catenatus Eastern Massasauga 

(=rattlesnake)

Threatened Reptiles

Sisyrinchium dichotomum White irisette Endangered Flowering Plants

Solidago houghtonii Houghton's goldenrod Threatened Flowering Plants

Solidago shortii Short's goldenrod Endangered Flowering Plants

Solidago spithamaea Blue Ridge goldenrod Threatened Flowering Plants

Somatochlora hineana Hine's emerald dragonfly Endangered Insects

Speoplatyrhinus poulsoni Alabama cavefish Endangered Fishes

Spigelia gentianoides Gentian pinkroot Endangered Flowering Plants

Spiraea virginiana Virginia spiraea Threatened Flowering Plants

Sterna dougallii dougallii Roseate tern Endangered Birds

Sternotherus depressus Flattened musk turtle Threatened Reptiles

Stygobromus hayi Hay's Spring amphipod Endangered Crustaceans

Thalictrum cooleyi Cooley's meadowrue Endangered Flowering Plants

Theliderma intermedia Cumberland monkeyface 

(pearlymussel)

Endangered Clams

Theliderma sparsa Appalachian monkeyface 

(pearlymussel)

Endangered Clams

Thelypteris pilosa var. 

alabamensis

Alabama streak-sorus fern Threatened Ferns and Allies

Torreya taxifolia Florida torreya Endangered Conifers and 

Cycads

Toxolasma cylindrellus Pale lilliput (pearlymussel) Endangered Clams

Trichechus manatus West Indian Manatee Threatened Mammals

Trillium persistens Persistent trillium Endangered Flowering Plants

Trillium reliquum Relict trillium Endangered Flowering Plants

Triodopsis platysayoides Flat-spired three-toothed Snail Threatened Snails

Tulotoma magnifica Tulotoma snail Threatened Snails

Vermivora bachmanii Bachman's warbler (=wood) Endangered Birds

Villosa fabalis Rayed Bean Endangered Clams

Villosa perpurpurea Purple bean Endangered Clams

Villosa trabalis Cumberland bean (pearlymussel) Endangered Clams
62

Appendix 2 LIST OF PREPARERS

Name, Title Education and Experience

Joanne Serrels

Biological Scientist

 M.S., Environmental Science & Policy, Johns Hopkins 

University.

 B.S., Wildlife Biology and Management, University of Rhode 

Island

 14 years of professional experience conducting NEPA

analyses.

 10 years of professional experience in environmental risk 

assessment of organisms produced through genetic 

engineering.

Scott Anderson

Biological Scientist-detail

 M.E.S., Environmental Studies, The Evergreen State College.

 B.S. Environmental Policy and Assessment, Western 

Washington University.

 15 years of professional experience conducting Endangered 

Species Act analyses, biological opinions, and NEPA

analysis.

Neil Hoffman

Science Advisor

 Ph.D. Plant Physiology, University of California, Davis.

 B.S. Plant Biology, Cornell University.

 20years of professional experience conducting Risk 

Assessment and NEPA analysis.

Lianne Hibbert

Assistant Chief, Biotechnology 

Environmental Analysis Services

Reviewer

 Ph.D., Human Dimension in Natural Resources, University of 

Missouri

 M.S., Fisheries and Wildlife Sciences, University of 

Missouri-Columbia

 B.S., Wildlife Biology, Grambling State University

 18 years of federal service and experience including policy 

development and review, developing program responses to 

congressional requests, and program management

 Ph.D., Human Dimension in Natural Resources, University 

of Missouri

 M.S., Fisheries and Wildlife Sciences, University of 

Missouri-Columbia

 B.S., Wildlife Biology, Grambling State University

 18 years of federal service and experience including policy 

development and review, developing program responses to 

congressional requests, and program management
63

Appendix 3 DISTRIBUTION LIST FOR THIS EIS

Distribution of this draft EIS to Contacts in EPA Regions:

Robert Koethe 

US EPA, Region 1 

5 Post Office Square, Suite 100 

Mail Code OEP06-3 

Boston, MA 02109-3912 

Grace Musumeci

US EPA, Region 2 

290 Broadway 

New York, NY 10007-1866 

Barbara Rudnick

US EPA, Region 3 

1650 Arch Street 

Philadelphia, PA 19103-2029 

Ntale Kajumba

US EPA, Region 4 

Federal Center

61 Forsyth Street, SW 

Atlanta, GA 30303-3104 

Ken Westlake

US EPA, Region 5 

77 West Jackson Boulevard 

Chicago, IL 60604-3507 

Michael Jansky

US EPA, Region 6 

Fountain Place 12th Floor, Suite 1200 

1445 Ross Avenue 

Dallas, TX 75202-2733 

US EPA, Region 7 

NEPA Program 

11201 Renner Blvd. 

Lenexa, KS 66219 

Philip Strobel

US EPA, Region 8 

1595 Wynkoop St. 

Denver, CO 80202-1129 

James Munson

US EPA, Region 9 

75 Hawthorne Street 

San Francisco, CA 94105 

Christine Reichgott

US EPA Region 10 

1200 Sixth Avenue, Suite 900 

Mail Code: ETTA-088 

Seattle, WA 98101 

____________________________________ 

No additional parties requested physical copies of the EIS. In addition to this distribution list 

APHIS notified all of its stakeholders of the availability of the EIS for review and comment.

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Appendix 5INDEX

acreage.............................................................................................................................. ix, 4-43, 4-44, 4-47

air quality ..............................................................................................................v, 3-7, 4-24, 4-40, 4-43, 72

alternatives ..............................................i, iii, iv, viii, ix, x, 3-4, 3-5, 4-34, 4-37, 4-39, 4-40, 4-41, 4-45, 4-46

climate change......................................................................................................... v, 3-13, 4-24, 4-36, 4-43

critical habitat ...................................................................................4-49, 4-50, 4-51, 4-52, 4-53, 4-54, 4-55

cross-pollination................................................................................................................................. vii, 4-38

economic environment.......................................................................................................iii, 2-1, 3-14, 4-39

endangered species....................................................................................................... 2-1, 4-49, 4-53, 4-55

environmental consequences..................................................................................................... vii, 3-4, 4-51

Federal Food, Drug and Cosmetic Act...................................................................................... ii, xi, 1-3, 4-33

Federal Insecticide, Fungicide and Rodenticide Act ................................................................ ii, xi, 1-2, 4-50

food and feed.......................................................................... 3-12, 3-15, 4-32, 4-33, 4-39, 4-44, 4-48, 4-55

Food Quality Protection Act................................................................................................................... ii, 1-2

gene flow.......................................................................................................................... 2-1, 3-9, 4-30, 4-52

greenhouse gases ............................................................................................................................... xi, 4-24

groundwater ........................................................................................................................v, 3-6, 4-23, 4-43

human health..............................................iii, 1-2, 2-1, 2-2, 3-12, 4-34, 4-35, 4-40, 4-41, 4-42, 4-44, 72, 76

isolation distances..................................................................................................................... vii, 3-15, 4-38

land use............................................................................................................................. ix, 4-43, 4-44, 4-47

National Environmental Policy Act ........................................ iii, ix, xi, 1-1, 1-4, 2-2, 4-39, 4-40, 4-42, 64, 65

pesticide registration .................................................................................................................................1-2

physical environment............................................................................................................................ iii, 1-4

plant diseases.............................................................................................................................................. 73

plant pest risk assessment........................................................................... iii, iv, xii, 1-1, 1-4, 3-4, 3-5, 4-42

plant pests...................................................i, ii, iv, 1-1, 1-2, 1-3, 3-4, 3-5, 4-45, 4-49, 4-50, 4-51, 4-52, 4-55

pollination .................................................................................... vi, 3-10, 3-15, 4-21, 4-29, 4-30, 4-31, 4-38

registered pesticide ...................................................................................................................................... ii

risk assessment................................................................................................................... 1-1, 1-3, 4-45, 64

scoping .................................................................................................................................................. iii, 2-1

socioeconomic impacts................................................................................................................................ vi

Toxic Substance Control Act ......................................................................................................................1-2

unreasonable adverse effects................................................................................................................ ii, 1-2

water resources ...................................................................................................... 3-7, 3-6, 4-22, 4-23, 4-40

weediness ........................................................................................................... 4-25, 4-42, 4-50, 4-51, 4-52

worker safety ...........................................................................................................................................4-47