NATIONAL ENVIRONMENTAL POLICY ACT DECISION 

AND 

FINDING OF NO SIGNIFICANT IMPACT 

Permit application 11-052-lOlrm 

received from ArborGen LLC 

Field testing of genetically engineered 

Eucalyptus grandis X Eucalyptus urophylla 

United States Department of Agriculture 

Animal and Plant Health Inspection Service 

Biotechnology Regulatory Services 

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

Service (APHIS) have developed a decision document to comply with the requirements of the 

National Environmental Policy Act of 1969, as amended, the Council of Environmental 

Quality's (CEQ) regulations implementing NEPA, and the USDA and APHIS NEPA 

implementing regulations and procedures. This NEP A decision document is intended to state 

APHIS' NEP A decision and present the rationale for its selection. 

In accordance with APHIS procedures implementing the NEPA Regulations (7 CFR part 372), 

APHIS has prepared an Environmental Assessment (EA) to evaluate and determine ifthere are 

any potentially significant impacts to the quality of the human environment in response to a 

permit application (11-052-101rm) received from ArborGen for the confined environmental 

release and interstate movement. If issued, the permit would authorize the planting of 

genetically engineered (GE) Eucalyptus hybrid clone (Eucalyptus grandis x Eucalyptus 

urophylla) to support proposed field research studies on six research sites in Alabama, Florida, 

Mississippi, and South Carolina. The permit application requests that APHIS permit all the trees 

except the trees on two of the sites in South Carolina to flower. The purpose of the confined 

environmental release is for ArborGen to assess the effectiveness of introduced gene constructs 

which are intended to confer cold tolerance; to test the efficacy of genes to alter lignin 

biosynthesis; to test the efficacy of genes to alter growth; and to test the efficacy of genes 

designed to alter flowering. In addition the trees have been engineered with a selectable marker 

gene which confers resistance to the antibiotic kanamycin. ArborGen has also requested under 

this permit application that all genetically engineered Eucalyptus trees authorized under permits 

09-070-101 rm and 11-041-101 rm be incorporated into this new permit and allowed to flower. 

New trees will also be moved interstate from ArborGen's nursery and greenhouse facilities 

located in South Carolina to the release locations identified in the permit application where they 

will be planted. 

The EA assesses the potential impacts of APHIS issuing or denying a confined environmental 

release and interstate movement permit with supplemental permit conditions to allow the 

interstate movement, planting, field testing and flowering of a GE Eucalyptus hybrid clone 

(Eucalyptus grandis x Eucalyptus urophylla) engineered to express various genes at 6 confined 

field site locations within in the States of Alabama, Florida, Mississippi, and South Carolina 

encompassing a total of 14.7 acres and analyzes the potential environmental and social effects 

that result from the proposed action and the alternatives. The proposed action of USDA APHIS, 

Page 1 of28 
Biotechnology Regulatory Services (BRS) is to issue the APHIS confined field release and 

movement permit for a hybrid of Eucalyptus grandis X Eucalyptus urophylla with supplemental 

permit conditions (see Appendices VI and VII of the EA). The movement authorization would 

be valid for one year and the release would be valid for a three-year period. Comments from the 

public involvement process were reviewed for substantive issues which were considered in 

developing this NEP A decision. 

Since 1986, the United States government has regulated GE organisms pursuant to a regulatory 

framework known as the Coordinated Framework for the Regulation of Biotechnology 

(Coordinated Framework) (51 FR 23302, 57 FR 22984). The Coordinated Framework, 

published by the Office of Science and Technology Policy, describes the comprehensive federal 

regulatory policy for ensuring the safety of biotechnology research and products and explains 

how federal agencies will use existing Federal statutes in a manner to ensure public health and 

environmental safety while maintaining regulatory flexibility to avoid impeding the growth of 

the biotechnology industry. The Coordinated Framework is based on several important guiding 

principles: (1) agencies should define those transgenic organisms subject to review to the extent 

permitted by their respective statutory authorities; (2) agencies are required to focus on the 

characteristics and risks of the biotechnology product, not the process by which it is created; and 

(3) agencies are mandated to exercise oversight of GE organisms only when there is evidence of 

"unreasonable" risk. 

The Coordinated Framework explains the regulatory roles and authorities for the three major 

agencies involved in regulating GE organisms: USDA's APHIS, the Food and Drug 

Administration (FDA), and the Environmental Protection Agency (EPA). 

APHIS is responsible for regulating GE organisms and plants under the plant pest provisions in 

the Plant Protection Act of2000, as amended (7 USC § 7701 et seq.) to ensure that they do not 

pose a plant pest risk to the environment. APHIS regulations at Title 7 of the Code of Federal 

Regulations (CFR) part 340, which were promulgated pursuant to authority granted by the Plant 

Protection Act, as amended (7 United States Code (U.S.C.) 7701-7772), regulate the 

introduction (importation, interstate movement, or release into the environment) of certain GE 

organisms and products. A GE organism is considered a regulated article if the donor organism, 

recipient organism, vector, or vector agent used in engineering the organism belongs to one of 

the taxa listed in the regulation (7 CFR 340.2) and is also considered a plant pest. A GE 

organism is also regulated under Part 340 when APHIS has reason to believe that the GE 

organism may be a plant pest or APHIS does not have information to determine if the GE 

organism is unlikely to pose a plant pest risk. 

When APHIS receives an application for a permit for environmental release and movement, the 

application is evaluated to determine whether the environmental release and movement, with 

appropriate conditions imposed, can be carried out while preventing the dissemination and 

establishment of plant pests. The receipt of a permit application to introduce a genetically 

engineered organism requires a response from the Administrator: 

Administrative action on applications. After receipt and review by APHIS of 

the application and the data submitted pursuant to paragraph (a) of this section, 

including any additional information requested by APHIS, a permit shall be 

granted or denied (7 CFR 340.4(e)). 

Page 2 of28 
The applicant has provided the required information associated with this request in the permit 

application. This information has been reviewed by APHIS-BRS and is analyzed in the EA. 

FDA regulates under the authority of the Federal Food, Drug and Cosmetic Act. The FDA 

policy statement concerning regulation of products derived from new plant varieties, including 

those genetically engineered, was published in the Federal Register on May 29, 1992 (57 FR 

22984-23005). Under this policy, FDA uses what is termed a consultation process to ensure that 

human food and animal feed safety issues or other regulatory issues (e.g., labeling) are resolved 

prior to commercial distribution of bioengineered food. The EPA regulates plant-incorporated 

protectants under the Federal Insecticide, Fungicide and Rodenticide Act (FIFRA) and certain 

biological control organisms under the Toxic Substances Control Act (TSCA). This GE 

Eucalyptus hybrid is not used for food or feed purposes and does not contain any genetically 

engineered pesticides or tolerance to herbicides. 

Public Involvement 

In a notice published in the Federal Register on February 10, 2012, (77 FR28 7123-7124, Docket 

No. APHIS-2011-0130), APHIS announced the availability of an EA for public review and 

comment for a proposed controlled field release of a genetically engineered clone of a 

Eucalyptus hybrid. Comments on the environmental assessment were required to be received on 

or before March 12, 2012. APHIS received a total of246 comments during the 30-day comment 

period. All respondents expressed concerns about the permit or opposed granting the permit. 

Comment documents may be viewed at 

http://www.regulations.gov/#!searchResults;rpp=25:po=O;s=APHIS%25E2%2580%25932011% 

25E2%2580%25930130. All comments were analyzed to identify new issues, alternatives, or 

information. Responses to the substantive comments are included as an attachment to this 

Finding ofNo Significant Impact. 

Major Issues Addressed in the EA 

The EA describes the alternatives considered and evaluated using the identified issues. Issues 

considered in the EA were developed based on APHIS' determination to issue a confined 

environmental release and interstate movement permit with supplemental permit conditions to 

allow the interstate movement, planting, field testing and flowering of a GE Eucalyptus hybrid 

clone (Eucalyptus grandis x Eucalyptus urophylla) engineered to express various genes at 6 

confined field site locations within the States of Alabama, Florida, Mississippi, and South 

Carolina. The following issues were identified as important to the scope of the analysis ( 40 CFR 

1508.25): 

• Interstate Movement of the trees under permit 

• Alteration in Susceptibility to Disease or Insects- Potential of the Eucalyptus to Harbor 

Plant Pests 

• Expression of the Gene Products, New Enzymes, or Changes to Plant Metabolism- Risk 

of the Gene Products on the Environment 

• Alteration in Weediness characteristics - Potential of the Engineered Eucalyptus to be 

Invasive 

• Possibility of Gene Flow Within the Field Test 

• Possibility of Gene Flow Outside of the Field Test 

• Possibility of Vegetative Propagation I Persistence Outside of the Field Test 

Page 3 of28 
• Potential of the Eucalyptus in the Field Tests to Become an Invasive Species that 

Threatens Native Plant and Animal Communities 

• Impact on Existing Agricultural Practices 

• Potential Impacts to Wildlife 

• Potential Impacts by Fire 

• Potential Impacts to Human Health 

• Transfer of Genetic Information to Organisms with which it Cannot Interbreed -

Horizontal Gene Transfer to Other Organisms 

• Potential Effects of Growing Eucalyptus on Soil Hydrology 

• Potential Allelopathic Effects of Eucalyptus 

• Risks to Threatened and Endangered Species 

• Impacts on Unique Characteristics of Protected Areas 

Affected Environment: 

The confined field tests would take place on land controlled by ArborGen or through private 

contracts established by ArborGen for field testing. The exact locations are claimed as CBI and 

have been submitted as part of the APHIS permit application, however the States and Counties 

where the confined field tests would occur are Baldwin and Escambia Counties, Alabama; 

Highlands County, Florida; Pearl River County Mississippi; and Berkeley and Dorchester 

Counties, South Carolina. Under the permit application submitted by ArborGen, there are six 

research sites where trees would be planted. A total of 14.7 acres (all 6 sites combined) is being 

requested by the applicant. Five of these locations currently have active APHIS permits (08-

0ll-106rm, 08-014-lOlrm, 09-070-lOrm, 10-112-lOlr, and 11-041-lOlrm) for environmental 

release of GE Eucalyptus hybrid clone (Eucalyptus grandis x Eucalyptus urophylla) in Alabama, 

Florida, Mississippi, and South Carolina. The remaining site in Dorchester County, South 

Carolina has been listed in previous APHIS permits/notifications as a holding area for transgenic 

trees. ArborGen is requesting that trees be allowed to flower at four locations in Alabama, 

Florida and Mississippi. At the two locations in Berkeley and Dorchester Counties, SC, they 

have requested to release trees in containers and have indicated they will not allow these trees to 

flower at these locations. The size of each individual confined field test sites identified in 

ArborGen's permit application ranges from 0.5 to 7.7 acres, which is less than the 20 acres 

analyzed in APHIS EA and subsequent FONSI prepared for Permits 08-0ll-106rm, 08-014-

lOlrm and 10-112-lOlr (http://www.aphis.usda.gov/brs/aphisdocs/06 325lllr ea.pdf) and 

(http://www .aphis.usda.gov /brs/aphisdocs/08 014101 r ea. pdf). For the five research sites that 

are currently authorized by APHIS to plant GE Eucalyptus hybrid clone (Eucalyptus grandis x 

Eucalyptus urophylla), ArborGen has indicated in their permit application that any new planting 

authorized by APHIS under 11-052-lOlrm would not expand the current field site locations 

beyond the areas currently authorized by APHIS. 

All the confined field test sites listed in the permit application by ArborGen are either on 

privately owned and managed plantation forests and agricultural farm lands or experimental 

research stations managed by academic institutions and industry, and have been in managed 

agricultural production for more than 5 years. The standard agricultural and silvicultural 

practices for land preparation, planting, irrigation, and harvesting of plants have been routinely 

used on these sites. Sites that include managed pastures have had intense activity including the 

use of heavy machinery for general upkeep, irrigation, fertilization, controlled grazing and 

management of grasses. Standard silvicultural practices would be used at these sites for the 

duration of the field tests. Surveys conducted by the applicant at each of these locations indicate 

Page 4 of28 
that there are not any old growth forests or undisturbed natural areas in the immediate 

surroundings of the test sites. ArborGen has indicated that the trees would be planted on 

individual research sites ranging from 0.5 up to 7.7 acres, depending on the location with 

planting density ranging from 300 - 600 trees per acre 

1

• 

The EA analyzes the potential environmental consequences of a proposal for APHIS to issue a 

confined environmental release and interstate movement permit with supplemental permit 

conditions to allow the interstate movement, planting, field testing and flowering of a GE 

Eucalyptus hybrid clone (Eucalyptus grandis x Eucalyptus urophylla) engineered to express 

various genes at 6 confined field site locations within the States of Alabama, Florida, 

Mississippi, and South Carolina. Based upon the permit application submitted by ArborGen, two 

alternatives are considered and analyzed in the EA: (1) deny the permit and (2) issue the APHIS 

permit. 

Alternative A: No Action- Deny the Permit 

Under the No Action alternative APHIS would deny the permit application (11-052-1 01rm) 

submitted by ArborGen. ArborGen would not be authorized to move and plant the GE 

Eucalyptus trees with new constructs and allow them to reach maturity and flower. Confined 

field release under permits 09-070-101rm and 11-041-101rm would continue to not allow 

flowering. 

There are currently six active permits under which ArborGen is authorized by APHIS to grow 

GE Eucalyptus which include 32 unique locations within 7 States: 08-011-106rm, 08-014-

101rm, 09-070-101rm, 10-112-101r, 11-041-101rm and 11-201-103r. No plantings at any ofthe 

32 locations are authorized by APHIS to exceed 20 acres in size. Trees are allowed to flower 

under four permits: 08-011-106rm, 08-014-101rm, 10-112-101r and 11-201-103r (two ofthe 

locations in Berkeley and Dorchester Counties, SC trees are not authorized to flower). Under 

permits 09-070-1 01 rm and 11-041-1 01 rm trees are not authorized to flower. As of September 

2011, ArborGen has reported to APHIS that they are growing a total of approximately 67 acres 

of trees on 18 of the 32 permitted locations. Under the No Action alternative, ArborGen could 

continue to grow GE Eucalyptus as authorized by APHIS under these six active permits. 

Alternative B: Preferred Alternative - Issue the APHIS Permit 

The Preferred Alternative is to approve ArborGen permit application request (11-052-1 0 1rm) 

and issue the APHIS confined field release and movement permit for a hybrid of Eucalyptus 

grandis X Eucalyptus urophylla with supplemental permit conditions (see Appendices VI and 

VII ofthe EA). The movement authorization would be valid for one year and the release would 

be valid for a three-year period. The permit will need to be renewed by ArborGen and 

subsequently approved by APHIS to allow the transgenic Eucalyptus plants to remain in the 

ground beyond the three-year time period specified in the permit application. 

Under this alternative, APHIS would issue a confined environmental release permit to ArborGen 

in accordance with 7 CFR part 340 to allow the movement, planting, field testing and flowering 

1 Planting density typically refers to the number of trees per acre. Planting densities can vary greatly depending 

upon the tree species and the environment, but densities of short rotation hardwood trees in the southeastern US 

are typically in the range of 30Q-800 trees per acre. Therefore sites ranging from 10 to 20 acres can have from 

3000 to 16,000 total trees planted in the ground. Twenty acres, as defined by forest plantation standards in the 

southeast, is considered a small planting. 

Page 5 of28 
of a GE Eucalyptus hybrid clone engineered to express various genes at 6 confined field site 

locations within in the States of Alabama, Florida, Mississippi, and South Carolina 

encompassing a total of 14.7 acres. The permit would allow all the trees except the trees on two 

of the sites in South Carolina to flower. All genetically engineered Eucalyptus trees authorized 

under permits 09-070-101rm and 11-041-101rm would be incorporated into this new permit and 

allowed to flower. This permit would authorize ArborGen to move and plant the GE Eucalyptus 

trees with new constructs and allow them to reach maturity and flower. Under this alternative, 

the applicant would be allowed to gather data on performance of the transgenic trees over a 

multiyear period and the efficacy of the genes in a wide variety of environments for multiple 

years. This alternative would allow the safe development and use of GE organisms under the 

mission ofBRS. 

Environmental Consequences of APHIS' Selected Action 

The EA contains a full analysis of the alternatives to which we refer the reader for specific 

details. The following table briefly summarizes the results for each of the issues fully analyzed in 

the Environmental Consequences section of the EA. 

Page 6 of28 
Table 1. Issue Alternative A Alternative B 

No Action Pre{j!rred Alternative 

Denv the Permit Issue APHIS Permit 

Interstate Movement of the trees under No increased risk No increased risk 

permit 

Alteration in Susceptibility to Disease or No increased risk No increased risk 

Insects - Potential of the Eucalyptus to 

Harbor Plant Pests 

Expression ofthe Gene Products, New No expected risk No expected risk 

Enzymes, or Changes to Plant Metabolism -

Risk of the Gene Products on the 

Environment 

Alteration in Weediness characteristics - Not likely Not likely 

Potential of the Engineered Eucalyptus to 

be Invasive 

Possibility of Gene Flow within the Field Minimal Minimal 

Test 

Possibility of Gene Flow Outside of the Minimal Minimal 

Field Test 

Possibility of Vegetative Propagation I Not likely Not likely 

Persistence Outside of the Field Test 

Potential of the Eucalyptus in the Field Not likely Not likely 

Tests to Become an Invasive Species that 

Threatens Native Plant and Animal 

Communities 

Impact on Existing Agricultural Practices No change No change 

Potential Impacts to Wildlife No adverse impacts No adverse impacts 

Potential Impacts by Fire Minimal risk Minimal risk 

Potential Impacts to Human Health Negligible Negligible 

Transfer of Genetic Information to No risk No risk 

Organisms with which it Cannot Interbreed 

- Horizontal Gene Transfer to Other 

Organisms 

Potential Effects of Growing Eucalyptus on No negative impacts No negative impacts 

Soil Hydrology 

Potential Allelopathic Effects of Eucalyptus Minimal Minimal 

Risks to Threatened and Endangered No effect No effect 

Species 

Impacts on Unique Characteristics of No effect No effect 

Protected Areas 

Page 7 of28 
Finding of No Significant Impact 

The analysis in the EA indicates that there will not be a significant impact, individually or 

cumulatively, on the quality ofthe human environment as a result of this proposed action. I 

agree with this conclusion and therefore find that an EIS need not be prepared. This NEP A 

determination is based on the following context and intensity factors (40 CFR 1508.27): 

Context- The term "context" recognizes potentially affected resources, as well as the location 

and setting in which the environmental impact would occur. This action would be limited to the 

interstate movement and environmental release of a GE Eucalyptus hybrid clone at 6 well 

dispersed confined field site locations in the States of Alabama, Florida, Mississippi, and South 

Carolina encompassing a total of 14.7 acres and has limited potential to affect resources outside 

of the confined field test sites. Supplemental permit conditions (see Appendices VI and VII of 

the EA) established for the permit will effectively limit the reproductive capabilities and 

establishment of this GE Eucalyptus outside the confined field trial locations and reduce the 

possibility of unintended exposure from moving GE Eucalyptus trees to be negligible to nonexistent. 

Intensity- Intensity is a measure of the degree or severity of an impact based upon the ten 

factors. The following factors were used as a basis for this decision: 

1. Impacts that may be both beneficial and adverse. 

According to the applicant, genetically engineered cold tolerant Eucalyptus would enable 

the production of this hardwood species for pulping and for biofuel applications in managed 

plantation forests in the southeastern U.S. APHIS issuance of the interstate movement and 

confined field release permit would allow ArborGen to conduct research to assess the 

efficacy of the introduced cold tolerance genes, genes to alter lignin biosynthesis in 

Eucalyptus, genes to alter growth and to research mechanisms for altered fertility. The 

confined release of the trees at six well dispersed confined field site locations in the States 

of Alabama, Florida, Mississippi, and South Carolina will allow the applicant to obtain data 

on performance of the transgenic trees and the efficacy of the inserted genes in a wide 

variety of environments. The establishment and growth of these small confined field tests 

will not have any impact on existing agricultural practices because they are solely for 

research purposes. Current agricultural practices will essentially remain unchanged. As 

identified by the applicant, the field sites that are being proposed under the permit have been 

used as forest tree plantations, pastures, or for forestry and agriculture research and are 

specifically designed for field testing crop plants or forest trees. As discussed in Section V 

of the EA, APHIS considers the possibility of unintended exposure from moving GE 

Eucalyptus trees under the proposed action to be negligible to non-existent. 

2. The degree to which the proposed action affects public health or safety. 

The proposed action to issue the APHIS interstate movement and confined field release 

permit should not pose an unnecessary risk to human health and therefore would have no 

significant impacts on human health. This GE Eucalyptus hybrid is not used for food or 

feed purposes and does not contain any genetically engineered pesticides or tolerance to 

herbicides. As described in Section V of the EA and in the response to comments, the risk 

that these field trials will result in a higher incidence of a fungal pathogen in the U.S. and 

thereby pose a risk to human health is considered to be negligible. 

Page 8 of28 
3. Unique characteristics of the geographic area such as proximity to historic or cultural 

resources, park lands, prime farmlands, wetlands, wild and scenic rivers, or ecologically 

critical areas. 

There are no unique characteristics of geographic area such as park lands, prime farm lands, 

wetlands, wild and scenic areas, or ecologically critical areas that would be significantly 

affected. As described in Section V and Appendix V of the EA, issuance of the permit to 

allow flowering of these additional constructs is not expected to impact unique 

characteristics of protected areas such as park lands, wetlands, wild and scenic areas, or 

ecologically critical areas. All the confined field test sites listed in the permit application 

are either on privately owned and managed plantation forests and agricultural farm lands or 

experimental research stations managed by academic institutions and industry. The standard 

agricultural and silvicultural practices for land preparation, planting, irrigation, and 

harvesting of plants have been routinely used on these sites. Surveys conducted by the 

applicant at each of these locations indicate that there are not any old growth forests or 

undisturbed natural areas in the immediate surroundings of the test sites. In addition, 

supplemental permit conditions (see Appendix VII of the EA) established for the permit will 

effectively limit the reproductive capabilities and establishment of this GE Eucalyptus 

outside the confined field trial locations and reduce the possibility of unintended exposure 

from moving GE Eucalyptus trees to be negligible to non-existent. 

4. The degree to which the effects on the quality of the human environment are likely to be 

highly controversial. 

The effects on the quality of the human environment are not highly controversial. Although 

there is some opposition to APHIS issuing the interstate movement and confined field 

release permit, this action is not highly controversial in terms of size, nature or effect. This 

action would be limited to 6 well dispersed confined field site locations in the States of 

Alabama, Florida, Mississippi, and South Carolina encompassing a total of 14.7 acres on 

individual research sites ranging from 0.5 up to 7.7 acres, depending on the location with 

planting density ranging from 300 - 600 trees per acre. During the public comment period, 

APHIS received comments opposing APHIS' issuance of the permit. No new issues, 

alternatives or substantive new information were identified in any of the comments received 

by APHIS. APHIS has addressed substantive comments in the response to public comments 

document attached to this FONSI based on scientific evidence found in peer-reviewed, 

scholarly, and scientific journals. 

5. The degree to which the possible effects on the human environment are highly uncertain or 

involve unique or unknown risks. 

The effects of the proposed action to issue the APHIS interstate movement and confined 

field release permit are not highly uncertain and do not involve unique or unknown risks. 

Based on the analysis documented in the EA, the effects on the human environment would 

not be significant. APHIS has no evidence for any unknown risks of this GE hybrid plant 

species when released for confined field testing or from moving GE Eucalyptus trees. This 

GE Eucalyptus hybrid has been previously released for confined field testing under APHIS 

issued notifications and permits in similar geographic locations without any known adverse 

effects. As described in Section V of the EA and response to comments, the issuance of the 

confined field release permit to allow the applicant to plant additional trees and to allow the 

trees to flower at four of the six locations does not present any unforeseen risks. Since there 

is no increase in the number of sites or acreage where trees will be allowed to reach maturity 

and flower, over those already authorized by APHIS to flower under permits 08-011-1 06rm, 

08-014-101rm, 10-112-101r and 11-201-103r with similar supplemental permit conditions, 

Page 9 of28 
APHIS has concluded that there is no substantial greater risk of loss of confinement and risk 

to the environment from the proposed action. As presented in Appendix I of the EA, 

information that has been collected by the applicant and provided to APHIS as part of 

ArborGen's annual reporting requirements under the existing permits that allow flowering 

confirmed that cold tolerant translines grown in these field tests did not produce any pollen, 

that the applicant has not observed any volunteer seedlings in or around the test sites where 

trees have flowered and produced seed capsules, and monthly field test monitoring 

observations have not identified any differences in diseases and insects or other non-target 

organisms between the transgenic and non-transgenic trees in the field test. Based on the 

analysis and information provided in the EA and supporting permit application, the new 

genes that are engineered into the trees should not pose any greater risk of loss of 

confinement and risk to the environment than those that are currently authorized by APHIS 

for confined field release. The addition of new genes to increase cold tolerance, alter lignin 

and growth should not compromise the ability of these to remain confined field tests. 

Adherence to the supplemental permit conditions established for the permit will effectively 

limit any potential adverse impacts to the human environment. As discussed in Section V of 

the EA, APHIS considers the possibility of unintended exposure from moving GE 

Eucalyptus trees under the proposed action to be negligible to non-existent. 

6. The degree to which the action may establish a precedent for future actions with significant 

effects or represents a decision in principle about a future consideration. 

The proposed action would not establish a precedent for future actions with significant 

effects or represent a decision in principle about a future decision. Similar to past permit 

applications reviewed and approved by APHIS, the decision on whether or not to issue a 

permit for interstate movement and confined environmental release will be based upon 

information provided in the permit application. APHIS regulations at 7 CFR part 340, 

regulate the introduction (importation, interstate movement, or release into the environment) 

of certain GE organisms and products. In accordance with these regulations, when APHIS 

receives an application for a permit for environmental release or movement, the application 

is evaluated to determine whether the environmental release or movement, with appropriate 

conditions imposed, can be carried out while preventing the dissemination and 

establishment of plant pests. The applicant has provided the information associated with 

this request in the permit application and APHIS now must make a determination to either 

grant or deny the permit. Each permit application that APHIS receives undergoes this 

independent review to determine if APHIS should grant or deny the individual permit. 

7. Whether the action is related to other actions with individually insignificant but 

cumulatively significant impacts. 

No significant cumulative effects were identified through this assessment. As discussed in 

the cumulative effects analysis presented in Section V of the EA, APHIS has determined 

that there are no past, present, or reasonably foreseeable actions that would aggregate with 

effects of the proposed action to create cumulative impacts or reduce the long-term 

productivity or sustainability of any of the resources (soil, water, ecosystem quality, 

biodiversity, etc.) associated with the release sites or the ecosystem in which they are 

situated. No resources will be significantly impacted due to cumulative impacts resulting 

from the proposed action. 

8. The degree to which the action may adversely affect districts, sites, highways, structures, or 

objects listed in or eligible for listing in the National Register of Historic Places or may 

cause loss or destruction of significant scientific, cultural, or historical resources. 

Page 10 of28 
The issuance of the interstate movement and confined field release permit would have no 

impact on districts, sites, highways, structures, or objects listed in or eligible for listing in 

the National Register of Historic Places, nor would they likely cause any loss or destruction 

of significant scientific, cultural, or historical resources. All the confined field test sites 

listed in the permit application are either on privately owned managed plantation forests and 

agricultural farm lands or experimental research stations managed by academic institutions 

and industry. The standard agricultural and silvicultural practices for land preparation, 

planting, irrigation, and harvesting of plants have been routinely used on these sites. Similar 

types of agricultural and silvicultural practices would be used by the applicant for managing 

trees associated with the permit. In addition, supplemental permit conditions (see Appendix 

VII of the EA) established for the permit will effectively limit the reproductive capabilities 

and establishment of this GE Eucalyptus hybrid outside the confined field trial locations and 

reduce the possibility of unintended exposure from moving GE Eucalyptus trees to be 

negligible to non-existent. 

9. The degree to which the action may adversely affect an endangered or threatened species or 

its habitat that has been determined to be critical under the Endangered Species Act of 

1973. 

APHIS evaluated the potential for negative effects on federal threatened and endangered 

species as listed by the U.S. Fish and Wildlife Service from the issuance of the interstate 

movement and confined field release permit and determined that the movement and 

confined environmental release of this GE Eucalyptus hybrid would have no effect on 

federally listed threatened or endangered species or species proposed for listing, or on 

designated critical habitat or habitat proposed for designation (see section on Threatened 

and Endangered Species in Section V and Appendix IV of the EA). 

10. Whether the action threatens a violation of Federal, State, or local law or 

requirements imposed for the protection of the environment. 

The proposed action would be in compliance with all federal, state, and local laws. The 

proposed action to issue the APHIS interstate movement and confined field release permit 

would be carried out in accordance with 7 CFR part 340. This GE Eucalyptus hybrid is not 

used for food or feed purposes and does not contain any genetically engineered pesticides 

or tolerance to herbicides. There are no other Federal, state, or local permits that are 

needed prior to the implementation of this action. 

NEPA Decision and Rationale 

I have carefully reviewed the EA prepared for this NEP A determination and the input from 

the public involvement process and have selected Alternative B-Issue the APHIS Permit. 

This alternative would fulfill APHIS' statutory mission and responsibilities to allow the safe 

development and use of genetically engineered organisms consistent with the plant pest 

provisions of the Plant Protection Act. 

As stated in the CEQ regulations, "the agency's preferred alternative is the alternative which the 

agency believes would fulfill its statutory mission and responsibilities, giving consideration to 

economic, environmental, technical and other factors." The preferred alternative has been 

selected for implementation based on consideration of a number of environmental, regulatory, 

and social factors. Based upon our evaluation and analysis, Alternative B is selected because (l) 

it allows APHIS to fulfill its statutory mission to protect America's agriculture and environment 

using a dynamic and science-based regulatory framework that allows for the safe development 

and use of genetically engineered organisms; and (2) it allows APHIS to fulfill its regulatory 

Page ll of28 
obligations. Therefore, it is my decision to implement the preferred alternative as described in 

the EA. Based on all of the analysis and reasons above, I have determined that there would be 

no significant impact to the quality of the human environment from the implementation of the 

chosen alternative (Alternative B) and therefore, no EIS needs to be prepared. 

Michael C. Gregoire 

Deputy Administrator Biotechnology Regulatory Services 

Animal and Plant Health Inspection Services 

U.S. Department of Agriculture 

Page 12 of28 

Date 
Attachment 

Finding of no significant impact 

Response to comments 

APHIS No. 11-052-lOlrm 

In a notice published in the Federal Register (77FR28 7123-7124, Docket No. APHIS-2011-

0 130), APHIS announced the availability of an EA for public review and comment for a 

proposed controlled field release of a genetically engineered clone of a Eucalyptus hybrid. 

Comments on the environmental assessment were required to be received on or before March 12, 

2012. APHIS received a total of246 comments during the 30-day comment period. All 

respondents expressed concerns about the permit or opposed granting the permit. Comment 

documents may be viewed at 

http://www .regulations.gov/#! searchResults;rpp=25 ;po=O;s= APHIS%25E2%25 80%259320 II% 

25E2%2580%25930 130. 

Respondents opposing APHIS' issuance of the permit came from nine non-governmental 

organizations (NGOs) or individuals apparently representing them, two State government 

entities, and individuals. The majority of respondents opposing the issuance of the permit came 

from one NGO, whose members submitted nearly identical form letters with some minor 

modifications. The NGO also submitted a petition bearing I ,957 signatories that opposed the 

issuance of the permit. The majority of respondents opposed believe that APHIS failed to 

comprehensively analyze all relevant issues related to the proposed field trial, that an 

Environmental Impact Statement should have been prepared and that cumulative impacts were 

not considered; that there is great potential for these Eucalyptus hybrids to become invasive or 

cross breed with non-transgenic Eucalyptus trees; Eucalyptus could be a host for Cryptococcus 

neoformans gattii, a fungi that could potentially cause mycoses in humans and animals; that 

Eucalyptus plantations have an impact on hydrology and cause or exacerbate drought situations; 

Eucalyptus trees are known to be an incendiary risk and tend to bum very hot during forest fires 

that are likely to occur in the drought prone southeastern U.S.; and could have significant 

impacts on biodiversity. APHIS' responses to these and all relevant comments submitted are as 

follows: 

Eucalyptus trees are introduced organisms in the U.S. and are documented as invasive pests 

in California and Florida. The cold tolerance trait could vastly expand the range of this 

GE Eucalyptus tree-and hence enhance its ability to invade native ecosystems. Experience 

in California and other parts of the world has clearly demonstrated that when Eucalyptus 

escape, it is next to impossible to eradicate them. Court decisions on genetically engineered 

perennial organisms including GE bentgrass and GE alfalfa, demonstrate a growing legal 

foundation around the potential escape of perennial GE organisms, even in field trials. 

The above comments were submitted in the form letter. 

Response: As covered in the EA, the genes introduced to affect cold tolerance could make the 

engineered Eucalyptus more adapted to cold temperatures in the southern United States, but this 

trait alone would not impart invasive or weediness characteristics to the engineered plants. The 

trees could be considered weedy or invasive if they were to produce many seedlings that were 

readily spread away from the field test sites. Where the non-engineered hybrid Eucalyptus 

(EH1) has been grown in Brazil, on an estimated 400,000 acres planted over 15 years, there has 

been no indication that large numbers of seedlings are being produced and are becoming invasive 

Page 13 of28 
from the commercial plantations. ArborGen is required to report any unusual occurrences, such 

as increased flowering or seed production to APHIS during the field testing period. 

The Eucalyptus being released is similar to the Veridian phytomining operation 

One commenter compared the release of the Eucalyptus under this permit to the Veri dian 

phytomining operation that released Y ellowtuft (Alyssum murale and A. corsicum) in southwest 

Oregon. The Alyssum escaped from cultivation and has been listed as a noxious weed in the State 

of Oregon. It is now a top priority for eradication in the State. 

Response: The situation under which these GE hybrid Eucalyptus trees would be grown and 

tested is very different from the situation that existed in Oregon under the Veridian project where 

the Y ellowtuft species were allowed to escape from cultivation. 

Y ellowtuft produces masses of yellow flowers and is a prolific seeder; and its height, from ten to 

thirty-six inches tall, makes it possible to harvest it by mechanical means. In addition, it is well 

adapted to the long, hot summers and relatively mild winters of southwest Oregon 

(http://www.oregonencyclopedia.org/entry/view/alyssum/). As discussed in the EA, the soil 

conditions in the southeastern U.S. are not conducive to germination and growth of Eucalyptus 

seedlings. The Alyssum species used in Oregon produces a large number of seedlings. The 

serpentine soils where they are being detected are highly conducive to the growth of the species. 

Supplemental permit conditions (see Appendix VII of the EA) established for the permit will 

effectively limit the reproductive capabilities and establishment of this GE Eucalyptus hybrid 

outside the confined field trial locations and reduce the possibility of unintended exposure from 

moving GE Eucalyptus trees to be negligible to non-existent. Under the supplemental permit 

conditions established for the permit ArborGen is required to monitor for any seedlings that are 

produced, remove them and report this to APHIS. The seedlings are easy to detect since they are 

distinct from other native species and they are readily controlled by the application of EPA 

approved herbicides. In addition, the GE hybrid Eucalyptus trees that would be grown in these 

confined field tests are similar to those already authorized by APHIS to flower under permits 08-

011-106rm, 08-014-1 01rm, 10-112-1 01r and 11-201-1 03r with similar supplemental permit 

conditions. As presented in Appendix I of the EA, information that has been collected by the 

applicant and provided to APHIS as part of their annual reporting requirements under the 

existing permits indicate no volunteer seedlings during the four years that they have been grown 

and allowed to flower. 

State of Florida Official-flowering of trees 

An official from the Florida Department of Agriculture and Consumer Services' Division of 

Plant Industry (DPI) expressed a concern that unlike other permits approved by DPI, these trees 

will be allowed to flower and that monitoring at the time of flowering and the subsequent 

information gathered at that time will be crucial to later decisions concerning the release of this 

cultivar on a larger scale. 

Response: APHIS contacted the official and discussed the misunderstanding about the permit 

with him. APHIS explained that GE Eucalyptus trees have been allowed to flower under permits 

in Florida since 2008 and that reproductive information associated with these permits has been 

collected by the applicant and provided to APHIS as part of their annual reporting requirements. 

To date no volunteers have been reported by the applicant. We also discussed the details of the 

supplemental permit conditions and explained that the permittee is required to scout for and 

Page 14 of28 
report the presence of any volunteer seedlings. We also explained that the individual research 

sites identified in the permit application would be similar in size to the confined field release 

permits that have been issued by APHIS in Florida. None of which have been over 20 acres in 

size. The official did not indicate that the agency had had any disagreement or concerns with the 

analysis provided in the EA. 

State of Georgia, District Supervisor Soil & Water District- potential invasive species 

This individual was concerned that the Eucalyptus that are being field tested could be another 

invasive species. He states that "until enough controlled tests are run and the American people 

can know for sure, USDA can control this invasive species, it should not be allowed to be 

massed planted." He indicates that he is not aware of how many years this has been field tested 

and under what conditions. 

Response: The permit application under consideration by APHIS is for the movement and 

confined field testing of GE Eucalyptus trees on a limited area. APHIS is considering issuing a 

confined environmental release and interstate movement permit with supplemental permit 

conditions to allow the interstate movement, planting, field testing and flowering of a GE 

Eucalyptus hybrid clone (Eucalyptus grandis x Eucalyptus urophylla) engineered to express 

various genes at 6 confined field site locations within the States of Alabama, Florida, 

Mississippi, and South Carolina encompassing a total of 14.7 acres. ArborGen has indicated that 

the trees would be planted on individual research sites ranging from 0.5 up to 7.7 acres, 

depending on the location with planting density ranging from 300 - 600 trees per acre. The GE 

hybrid Eucalyptus trees that would be grown in these confined field tests are similar to those 

already authorized by APHIS to flower under permits 08-011-1 06rm, 08-0 14-101 rm, 1 0-112-

101r and 11-201-103r with similar supplemental permit conditions. As presented in Appendix I 

of the EA, information that has been collected by the applicant and provided to APHIS as part of 

their annual reporting requirements under the existing permits have produced no volunteer 

seedlings during the four years that they have been grown and allowed to flower. As reported by 

the applicant, these cold tolerant translines grown in these field tests did not produce any pollen, 

the applicant has not observed any volunteer seedlings in or around the test sites where trees 

have flowered and produced seed capsules, and monthly field test monitoring observations have 

not identified any differences in diseases and insects or other non-target organisms between the 

transgenic and non-transgenic trees in the field test. Based on the analysis and information 

provided in the EA and supporting permit application, the new genes that are engineered into the 

trees should not pose any greater risk of loss of confinement and risk to the environment than 

those that are currently authorized by APHIS for confined field release. The addition of new 

genes to increase cold tolerance, alter lignin and growth should not compromise the ability of 

these to remain confined field tests. Adherence to the supplemental permit conditions 

established for the permit by the applicant will effectively limit any potential adverse impacts to 

the human environment. As discussed in Section V of the EA, APHIS considers the possibility 

of unintended exposure from moving GE Eucalyptus trees under the proposed action to be 

negligible to non-existent. Nothing in the comment indicates that there was any disagreement 

with the analysis provided in the EA. 

Genetic engineering alters genomes in a random and unpredictable fashion 

One commenter cited a paper by Latham (2006) which says that "Despite the fact that 

confidence in the safety and dependability of crop species rests significantly on their genetic 

integrity, the frequency of transformation-induced mutations and their importance as potential 

Page 15 of28 
biosafety hazards are poorly understood." The commenter goes on to state that "Releasing GE 

trees will mean releasing trees that have never existed before with totally unknown qualities. 

This is damaging to biodiversity and since they are living organisms this pollution is 

uncontainable and has the potential to grow over time." 

Response: The engineered Eucalyptus trees in the proposed confined field tests have been 

observed over the course of nine years in the laboratory, greenhouse and in confined field tests to 

identify any unusual or unintended effects due to the transformation process. As confirmed in 

field tests conducted by the applicant since 2006, only the traits for which the trees have been 

transformed have been expressed in these trees. In most cases any transformation-induced 

mutations would have been eliminated early in the transformation process as trees were 

regenerated in the laboratory and grown for field testing. The permit application under 

consideration by APHIS is for the movement and confined field testing of GE Eucalyptus trees 

in a limited area. APHIS is considering issuing a confined environmental release and interstate 

movement permit with supplemental permit conditions which includes specific monitoring and 

reporting requirements. In addition, APHIS would also conduct yearly inspections to look for 

unintended effects. The GE hybrid Eucalyptus trees that would be grown in these confined field 

tests are similar to those already authorized by APHIS to flower under permits 08-011-1 06rm, 

08-014-101 rm, 10-112-101 r and 11-201-1 03r with similar supplemental permit conditions. As 

presented in Appendix I of the EA, information that has been collected by the applicant and 

provided to APHIS as part of their annual reporting requirements under the existing permits have 

not identified any differences in diseases and insects or other non-target organisms between the 

transgenic and non-transgenic trees in the field test. Given the small size of the confined field 

test sites identified in the permit application and the supplemental permit conditions that would 

be imposed on the applicant and the fact that the permits can be revoked at any time by APHIS 

and the trees removed if any unintended effects are found, APHIS does not anticipate that 

issuance of the confined environmental release and interstate movement permit would impact 

biodiversity. 

Excessive claims of CBI and failure of ArborGen to reveal critical information 

A couple of commenters indicated that the designation of the majority of transgenic 

modifications as CBI prevents any rational evaluation of the impact of those genes and modified 

Eucalyptus on the environment, humans and animals. And because the locations are claimed as 

CBI it is impossible to create a creditable environmental assessment for the permit application. 

Also a commenter indicated that normally consideration would be limited to only those species 

and critical habitat identified by the United States Fish and Wildlife Service and/or National 

Marine Fisheries Service likely to be found in the area of the field tests. The commenter 

believed that, without this information, such consideration is impossible. There was a concern 

that "bystanders exposed to the CBI GM Eucalyptus pollen, dust from decaying leaf detritus and 

pollution of surface and ground water from polluting plant products" would suffer adverse side 

effects. 

Response: APHIS takes the use and designation of CBI very seriously. Like all Federal 

agencies, APHIS is subject to the Trade Secrets Act (18 U.S.C. § 1905) (TSA). The TSA 

prohibits officers and employees of federal agencies from publishing or disclosing trade secrets 

and other confidential business information "to any extent not authorized by law." Information 

that is claimed as CBI by applicants is evaluated by APHIS to ensure that the claims are 

legitimate and APHIS evaluated ArborGen's claims for CBI protection in this instance. It would 

Page 16 of28 
be a clear violation of the Act for APHIS to reveal Confidential Business Information to the 

public that legitimately falls under the Act. Federal employees can be fined and/or imprisoned 

for violating the TSA. Every gene construct component in the permit submission is evaluated for 

any plant pest risk as well as any potential effects on the environment. None of the genes that 

have been engineered into these trees (to affect cold tolerance, lignin formation, growth and 

pollen formation) are expected to have any effects on "bystanders exposed" to the Eucalyptus 

trees or potentially pollute surface and ground water. No pollen is being produced by the trees in 

the field tests due to the presence of the barnase gene. In addition a thorough analysis for 

threatened and endangered species and critical habitat are made for every location and if APHIS 

determines that there could be a species present at the field test site, appropriate contacts are 

made with the USFWS. A thorough discussion of potential impacts of the proposed action on 

human health, water resources and threatened and endangered species are presented in Section V 

and Appendix IV of the EA. 

Concern that the CBF genes could have pleiotropic effects and affect allelopathy 

One commenter indicated that the C-Repeat Binding Factor (CBF) genes when over expressed 

display improvement in cold tolerance, water retention, higher oil gland density, wax deposition 

and over expression of anthocyanin pigments (Navarro et al., 2011 ). The gene network 

influenced by CBF gene modification produced highly pleiotropic. The commenter was 

concerned that the production of allelochemicals in the modified trees did not appear to have 

been investigated. 

Response: The reference cited in the comment (Navarro et al., 2011) does indicate that the gene 

can have multiple phenotypic effects. As a transcription factor this is not unexpected. As the 

publication indicates, the multiple changes observed are consistent with the correlation between 

cold tolerance and drought tolerance and reinforce the hypothesis that, by triggering metabolic 

changes directed towards limiting cell dehydration, the DREB transcription factors are partly 

responsible for these observed correlations. As covered in the EA, allelopathy tends to be an 

inexact science and many studies in allelopathy are inconclusive and difficult to interpret due to 

potential interactions with other aspects of the environment. Even though the Eucalyptus under 

this permit could demonstrate allelopathic properties, the presence of any allelochemicals is not 

going to make the Eucalyptus planted under these permits more invasive or present a plant pest 

risk. Since all these field tests are confined and limited in size, any allelopathic effects should be 

small. As a standard silvicultural practice, herbicides will also be used within the field test sites 

and any of their effects on understory vegetation will be as severe or more severe than any 

allelopathic effects. 

Pollen spread 

One commenter was concerned about pollen spread from the trees and cites an Australian study 

that found that remnant populations of Eucalyptus were connected by pollen dispersal to pollen 

sources up to 1.94 kilometers away (Sampson and Byrne, 2008). The concern is that the 

distance between these GE Eucalyptus and other Eucalyptus that are 1000 meters away would 

lead to the spread of the GE trees. 

Response: As indicated in the EA, the observations made to date by the applicant on APHIS 

approved confined field tests that have allowed GE Eucalyptus hybrid trees to flower indicate 

that the flowering of the hybrid does not overlap the flowering of other Eucalyptus in Florida 

including one ofthe parents of the hybrid, i.e. E. grandis. Also given that the transgenic hybrid 

Page 17 of28 
is not producing any pollen, there is little if any potential for outcrossing of the hybrid to other 

species. One of the requirements of the existing permits and proposed permit is that the 

applicant must report any overlap in flowering if it occurs with other Eucalyptus tree species in 

the area. If any overlap in flowering were to occur, the only seeds that could be formed from 

crossing would be on trees within the test plots whose female flowers could be susceptible to 

incoming pollen. Monitoring of any seeds and seedling formation in the permitted plots by the 

applicant would show if any seedlings were being formed as hybrids from incoming pollen 

sources. In accordance with the supplemental permit conditions, monitoring for and removal of 

volunteers within lOOm from the edge oftransgenic test plot by the applicant would effectively 

eliminate any seedlings that may be produced and thereby effectively reduce the possibility that 

any Eucalyptus hybrid trees would become established. 

Concerns about Cryptococcus 

One commenter and the form letter was concerned about the spread of Cryptococcus neoformans 

gattii by the transgenic Eucalyptus trees indicating that the spread of this "deadly yeast" is 

mainly brought about by birds and their feces. The commenter appears to take issue with the 

statement in the EA that the scale of the field tests is miniscule compared to the vast expanses of 

native trees that could potentially harbor the pathogen and says that "There was no vast expanse 

of native species homes for the toxic yeast in the peer reviewed scientific publications." 

Response: As covered in the EA, it is well documented in the literature that many trees, other 

than Eucalyptus, can harbor Cryptococcus in North America. According to the APHIS report 

cited in the EA (APHIS, 2004): "In the endemic area of Vancouver Island, trees that have tested 

positive for C. gattii include alder, bitter cherry, cedar, Douglas Fir, and Garry Oak." Many 

years after its first recovery from clinical specimens, C. gattii was isolated from the environment 

for the first time by sampling of Eucalyptus camaldulensis trees (Ellis and Pfeiffer, 1990). This 

tree species was initially thought to be the exclusive natural habitat of C. gattii both in its native 

Australia and around the world given that the tree has been exported extensively. However now 

it is clear that more than 50 tree species have been reported as harboring the fungus (Springer 

and Chaturvedi, 201 0). Although Eucalyptus is present in many of the areas known to have C. 

gattii-associated cryptococcosis, the actual isolation of C. gattii from Eucalyptus trees is rare 

outside Australia, despite extensive sampling. Imported Eucalyptus has not been associated with 

the environmental presence of C. gattii in Spain, central Africa, or Canada; and most Eucalyptus 

trees tested in Papua New Guinea, Egypt, and Italy were negative for C. gattii. When C. gattii 

was first associated with cryptococcosis, early environmental surveys for C. gattii in imported 

Eucalyptus spp. rarely included other local tree species for testing. Although understandable, 

this was unfortunate because C. gattii is now known to have extensive associations with other 

tree species (Springer and Chaturvedi, 201 0). 

C. gattii is clearly not specific to Eucalyptus trees in North America and other areas of the world 

and is most likely being spread by birds and other animals (Springer and Chaturvedi, 201 0) 

throughout temperate tree species since very little Eucalyptus is present in these areas. 

Therefore, the planting GE Eucalyptus trees in confined field tests in southeast would not be 

expected to increase the incidence of the disease any more than planting any other tree species, 

since many species can host the fungus. Also it is important to note that the recent literature has 

begun to sort out the difference between C. neoformans and C. gattii. Until recently, these two 

fungi were classified as varieties of the same species but are now considered two different 

spectes. The pathogen C. gattii is believed to be clinically more virulent than C. neoformans and 

Page 18 of28 
causes more severe symptoms of cryptococcosis. For a review see (Chaturvedi and Chaturvedi, 

2011). 

Concerns about the alteration in susceptibility to diseases or insects 

Commenters were concerned that the susceptibility of the trees to insects and disease were not 

adequately addressed in the EA, particularly the trees that have altered lignin. 

Response: The GE hybrid Eucalyptus trees that would be grown in these confined field tests are 

similar to those already authorized by APHIS to flower under permits 08-011-1 06rm, 08-014-

101rm, 10-112-101r and 11-201-103r with similar supplemental permit conditions. As presented 

in Appendix I of the EA, information that has been collected by the applicant and provided to 

APHIS as part of their annual reporting requirements under the existing permits have not 

identified any differences in diseases and insects or other non-target organisms between the 

transgenic and non-transgenic trees in the field test. Growth measurements have indicated that 

trees had normal to a moderately reduced growth phenotype. The trees have been visually 

inspected on a monthly basis for the presence of any insect and disease damage. As reported in 

the applicant's annual reports, the degree of insect and disease infection has been minimal. 

Adherence to the supplemental permit conditions established for the permit by the applicant will 

effectively limit any potential adverse impacts to the human environment. However, if during 

the tests there is evidence of increase disease or insect susceptibility, the applicant is required to 

report this to APHIS. The permittee is required to report any such unanticipated effects 

(including excessive mortality or morbidity) to APHIS under the terms of the permit- see 7 CFR 

340.4(f)(1 O)(ii). An assessment of disease and insect susceptibilities is one of the primary 

reasons the applicant is conducting the confined field testing of the trees. 

Kanamycin resistance 

One commenter raised an issue about the risks from the kanamycin resistance gene. The 

commenter indicated that kanamycin is still in clinical use and also kanamycin resistance crossreacts with new antibiotics. 

Response: In 1998, FDA developed draft guidance on the use of antibiotic resistance markers in 

transgenic plants 

http://www.fda.gov/food/guidancecomplianceregulatoryinformationlguidancedocuments/biotech 

nology/ucm096135.htm. In this document, FDA concluded that the likelihood of the transfer of 

antibiotic resistance is remote. In this draft guidance the FDA stated, "FDA acknowledges that 

the likelihood of transfer of an antibiotic resistance marker from plants to microorganisms in the 

gut or in the environment is remote and that, such transfer, if any, would likely be insignificant 

when compared to transfer between microorganisms, and in most cases, would not add to 

existing levels of resistance in bacterial populations in any meaningful way." Further, to 

evaluate safety, the European Food Safety Authority (EFSA) reviewed the antibiotic selection 

markers used in genetically engineered plants 

http://www.efsa.europa.eu/enlefsajoumal/doc/opinion gmo 05 en1.pdf. In this 2004 document, 

various antibiotic resistance genes were assigned into groups based on the criteria of therapeutic 

use in humans and in animals and presence in the environment; Group I is composed of 

kanamycin and hygromycin resistance genes. The opinion states that because of the frequency 

of horizontal transfer plants to other organisms is very rare, previous existence in the 

environment and the history of use of the kanamycin resistance, that there is no rationale for 

restricting Group I antibiotics. In addition, the EFSA 2004 document states: Kanamycin is 

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rarely used today because of its considerable side effects. Only under conditions of multiple 

mycobacterial resistance to other drugs is kanamycin still used as a reserve tuberculostatic agent. 

For the same reason as kanamycin, neomycin, which is poorly absorbed orally, is also rarely 

used intravenously/intramuscularly to treat infections. These antibiotics are rarely administered 

orally, which minimizes the selective pressure for antibiotic resistance in the gut. Their use in the 

treatment of humans has been superseded by more effective amino glycoside antibiotics that are 

not substrates for APH(3')-II. The antibiotics are rarely used in agriculture or aquaculture and 

thereby do not provide selective pressure for a possible transfer of the resistance genes from 

genetically modified plants to soil microorganisms. 

Gene for altered fertility 

Commenters were concerned about the "dangers of the barnase gene" and stated that it is a 

known cytotoxic protein that breaks down RNA. "Because they are from a soil bacterium, and 

unrelated to any mammalian RNAses, they are not susceptible to eukaryotic 3 RNAse inhibitors. 

Consequently, they are highly toxic, and are actually being engineered currently as a means of 

killing cancer cells. Although the barnase is being used to prevent pollen formation, this is not 

100 [%]effective, and many beneficial insects and other wild-life could well be affected." 

Response: As indicated in the EA, the transgenic Eucalyptus hybrid identified in the permit 

application utilizes barnase to produce male sterile plants, reducing the potential for gene flow 

into the environment. Male sterility is achieved through the localized production of barnase in 

pollen producing cells. Barnase is a ribonuclease, an enzyme that degrades RNA, thereby 

regulating protein synthesis. Ribonucleases are highly ubiquitous molecules found in all living 

cells (Worrall and Luisi, 2007). The transgenic Eucalyptus trees identified in the permit 

application and other permits already authorized by APHIS were engineered using barnase gene 

mutants with reduced toxicity to minimize tissue damage (Rottmann et al., 2008). The barnase 

production is controlled by a tissue specific promoter. As with all genes, the gene encoding 

barnase is present in every cell of the transgenic plant, however, the promoter acts like an "on" 

switch that controls when and how strongly the gene is expressed. The PrMC2 promoter used by 

ArborGen, originally identified in pine (Pinus radiata), restricts barnase production to the 

tapetum, a small layer of cells within the male floral organ, or anther (Walden et al., 1999; Hofig 

et al., 2003; Rottmann, et al., 2008). Within the anther, the tapetum surrounds developing pollen 

grains. Mariani, et al. (1990) developed transgenic tobacco plants using the barnase gene also 

controlled by a tapetum-specific promoter (Tap29). They observed that in transgenic plants 

tapetal cells senesced early in their development, preventing pollen formation. All other floral 

organs, including the anthers, formed normally proving tapetal cells which could be eliminated 

without affecting flower development. Similar results were observed using PrMC2 in Eucalyptus 

trees (Rottmann et al., 2008). After several years of research, no pollen has been observed in 

transgenic Eucalyptus trees as reported in the permit application and in annual reports submitted 

by ArborGen up through 2011 (see Appendix I of the EA). As with other ribonucleases, barnase 

degrades quickly after the destruction of tapetal cells, and does not accumulate within the plant 

tissues (Mariani et al., 1990); http://www.fda.gov/Food/Biotechnology/Submissions 

/ucml61162.htm). Therefore barnase is only produced for a short period of time during floral 

development, is limited to a discrete cell layer within the anthers of the flower, and is rapidly 

degraded (Mariani et al., 1990; Rottmann et al., 2008; Hofig et al., 2003). In previous studies 

using transgenic radicchio containing the barnase gene, researchers were unable to detect 

accumulation of barnase within floral tissue 

(http://www.fda.gov/Food/Biotechnology/Submissions/ucml61162.htm). Therefore it is highly 

Page 20 of28 
unlikely that consumption of, or exposure to, transgenic Eucalyptus could contain enough 

bamase to cause mammalian or insect toxicity. Direct exposure of organ tissue, is also highly 

unlikely to occur in nature. In addition the FDA has previously reported that consumption of 

bamase would likely degrade quickly during digestion further reducing the risk of bamase 

exposure (http://www.fda.gov/Food/Biotechnology/Submissions/ucm 161162.htm). 

With regard to toxicity of bamase to bees and other pollinating insects, there is no clear evidence 

indicating pollinators would be adversely affected by ingestion ofbamase. Combined with the 

facts that bamase is only produced during the short period of tapetum formation and is quickly 

degraded, as well as the lack of pollen produced by the Eucalyptus hybrids, it can be concluded 

the tissue specific production of barnase is unlikely to adversely affect pollinators or other 

insects. APHIS has made a determination of nonregulated status of three plant species, com 

(petitions 95-288-01p, 97-342-01p, 98-349-01p), rapeseed (petitions 98-278-01p, 01-206-01p) 

and chicory (petition 97-148-01p) since 1995 that have included the barnase gene. APHIS is 

unaware of any reported cases of mammalian or insect toxicity resulting from barnase 

consumption or exposure occurring within the past 17 years. 

Based on these findings, no hazard or exposure associated with the use of barnase has been 

identified, therefore APHIS concludes that there is no foreseeable risk associated with the use of 

bamase in transgenic Eucalyptus trees in the proposed confined field tests. 

Mode of transformation and hazards of horizontal gene transfer 

One commenter expressed concerns about horizontal gene transfer between the Agrobacterium 

used to transform the plants to wild-type Agrobacterium in the soil, and to other bacteria and 

fungi in the soil, citing a publication by (Knight, 201 0). 

Response: It is highly unlikely that any Agrobacterium remains on the plants that are generated 

in the laboratory since antibiotics are used to remove them prior to release into soil. During the 

Eucalyptus transformation process, the transformed tissue is kept on selection medium 

containing antibiotics to kill any residual Agrobacterium for at least four months, following 

which individual events are transferred to medium without selection. As indicated by the 

applicant, Agrobacterium growth has never been observed in the cultures at this stage. 

ArborGen has also collected vegetative tissue from select tissue cultures as well as field-grown 

transgenic plants and analyzed for the VirD gene which would indicate the presence of 

Agrobacterium. The lack of detectable VirD signal in samples indicates that no live or dormant 

Agrobacterium remains in the transgenic plants. 

High flammability of Eucalyptus 

Several commenters including the form letter noted the high flammability of Eucalyptus which 

allows Eucalyptus plantations to be susceptible to initiation or rapid spread of wildfire. 

Response: As indicated in the EA, it is not clear that Eucalyptus plantations present a greater 

fire hazard than do the pine plantations which are common in the Southeastern USA. To assess 

the risk of wildfire in live and dead material, (Nufiez Regueira et al., 2002a; Nufiez Regueira et 

al., 2002b) in northern Spain used calorimetry analysis combined with multidimensional 

assessment of climatic and physical characteristics in plantations of a Pinus species (P. pinaster) 

and of a Eucalyptus species (E. globulus). For pine, the fire risk index for live biomass varied 

from extremely high in July to little risk in some winter and spring months. Pine residues, 

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however, varied between no risk and extremely high risk. For Eucalyptus, fire risk index for live 

biomass varied from a middle risk in July to little risk in fall and winter. Dead matter of 

Eucalyptus varied between a middle risk to extremely high risk in August. Ganteaume et al. 

(2009) undertook a variety of assays and comparisons of fuel bed flammability and firebrand 

capacity (the material that can ignite the fuel bed). Pinus litter flammability was higher than that 

of Eucalyptus, which in tum was higher than that of a hardwood species (Ulex). While 

differences between southeastern pines and the proposed Eucalyptus species and these analyses 

may be important, basic similarities within the genera are likely relevant. For example, studies 

of different Eucalyptus species have shown that despite the differing climactic specificities of E. 

globulus and E. nitens and differences of volatile content and solid mass, the total caloric content 

of wastes after pulping are similar (Perez et al., 2006). Due to the small size of these field trials, 

and the fact that they are isolated from one another by many miles, there is no reason to believe 

that they pose any more of a fire risk than do other forest tree plantings 

Wildfires in California and Australia 

Some of the comments and the form letter cited two large wildfires in arid areas of Australia and 

California involving Eucalyptus. These fires were mentioned as examples of deleterious 

consequences of Eucalyptus plantings. 

Response: These fires occurred in areas of extensive plantings of Eucalyptus, but it is likely that 

many factors are important for the frequency and season of occurrence of fires, not just the 

species of trees within a burned area. One post-fire report on the Oakland, California fire noted 

that extended drought conditions and freeze-killed Eucalyptus contributed large amounts of 

residue for the fire. However, a wide variety of grasses, brush, trees, and houses combined to 

provide fuel that sustained this fire, and the fire had not initiated in the Eucalyptus stands 

(FEMA, 1991). As noted, one ofthe most important risk factors for Eucalyptus production is the 

presence of accumulating fuels within a stand (Nufiez Regueira, et al., 2002a). The duration of 

these field trials will be limited (no more than seven to nine years). ArborGen has seen little 

accumulation in field trials extending through five years (Les Pearson, ArborGen, personal 

communication) and consequently little to no accumulating fuel is expected to arise within the 

proposed confined field trials. 

Failure to Consider the Environmental Effects of Eucalyptus grandis X Eucalyptus 

urophylla Pollen on Bee Populations 

One commenter was concerned that the EA did not consider the effect of pollen on bee 

populations. There was a concern that when the flower pollen becomes genetically modified or 

sterile, the bees will potentially go malnourished and die of illness due to the lack of nutrients 

and the interruption of the digestive capacity of what they feed on through the summer and over 

the winter hibernation process. He also indicates that there already exist sufficient acreage and 

plots to capture and test pollen from GMO Eucalyptus and to monitor its effect on bee 

populations in proximity to them. And if this process is shown to have no deleterious effects on 

existing bee populations, over a series of years should any proposal to expand the range of GMO 

Eucalyptus be considered. 

Response: It is important to note that the trees are not producing any pollen, therefore the 

studies that are proposed by the commenter cannot be performed. Also, as discussed above and 

within the EA, there is no reason to believe that the products of the introduced genes would have 

any negative effects on insects such as bees. ArborGen has observed many bees visiting the 

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Eucalyptus flowers in the field test because the flowers are very attractive to bees and they feed 

on the nectar produced by the flowers. As noted above no products (including barnase) produced 

by the inserted genes would be expected to affect bees. No dead bees have been observed in the 

vicinity of the field tests. 

Unacceptable Environmental Risks: Water Resources 

Several comments including the form letter noted the potential impacts of Eucalyptus on 

hydrology. The comments centered on concerns that tree plantations have been documented to 

deplete ground water and cause or exacerbate drought situations. They also cited the information 

that the Forest Service provided upon APHIS' request as an appendix to the EA to indicate that 

the risk is unacceptable. 

Response: The proposed action would be limited to 6 well dispersed confined field site 

locations in the States of Alabama, Florida, Mississippi, and South Carolina encompassing a total 

of 14.7 acres on individual research sites ranging from 0.5 up to 7.7 acres. ArborGen has 

supplied data to APHIS indicating the maximum size of each of the plantings at each site, the 

individual watersheds where the plantings occur, the area of the watershed, how much of the 

watershed will be occupied by the field tests, the location of the closest primary and secondary 

streams, and the location of any critical habitat for Federally listed threatened and endangered 

species within the watershed. The data provided by ArborGen show that none of the potential 

planting sites identified in the permit application occupy more than 0.005% of any given 

watershed. 

The potential impacts by Eucalyptus on hydrology relate to the scale of the plantings. The 

proposed confined field trial plantings are very small in terms of scale in forestry practices and 

watersheds. Individual forest plantings in the southern U.S. range in size but can typically be up 

to 120 acres at a single site in a given year (SFI standards: 

http://www.sfiprogram.org/files/pdf/sfi requirements 2010-20 14.pdf). A 20 acre planting is 

considered small in these terms. The issue of hydrology is thoroughly addressed in Section V of 

the EA. As noted in the section under Potential Effects of Growing Eucalyptus on Soil 

Hydrology in the EA, the Forest Service has pointed out that the significance of the impact on 

groundwater and stream flow will depend greatly on the area extent, size, and spatial distribution 

of the plantations. For example, a few small (less than 10 hectares, i.e. approximately 25 acres) 

and well-dispersed plantations may only have very localized impacts and negligible impacts at 

the watershed scale. Based on the very small footprint of these proposed confined field tests and 

the weight of evidence, APHIS concludes that the impacts of these field trials on hydrology will 

be negligible. 

Unacceptable environmental risks: Native Flora and Fauna 

One commenter noted a comment made on a previous EA for GE Eucalyptus permits. The 

commenter states that "The Georgia Department ofNatural Resources, Wildlife Resources 

Division has recommended that ArborGen's permit requests be denied because the Department 

has "serious concerns about potential impacts on hydrology, soil chemistry, native biodiversity 

and ecosystem functions regardless of whether this nonnative hybrid turns out to be invasive in a 

plantation setting." They further note that Eucalyptus plantations "will be extremely inhospitable 

environments for native flora and fauna." Moreover, noting the high water use for Eucalyptus 

plantations, the Department expressed concern regarding the increased potential for significant 

impacts on water resources and aquatic communities. 

Page 23 of28 
Response: The potential impacts of the proposed action on native flora and fauna is thoroughly 

analyzed in Section V of the EA and discussed in several of the responses above. 

The GE hybrid Eucalyptus trees that would be grown in these confined field tests are similar to 

those already authorized by APHIS to flower under permits 08-011-1 06rm, 08-014-101 rm, 10-

112-101r and 11-201-1 03r with similar supplemental permit conditions. As presented in 

Appendix I of the EA, information that has been collected by the applicant and provided to 

APHIS as part of their annual reporting requirements under the existing permits have produced 

no volunteer seedlings during the four years that they have been grown and allowed to flower. 

As reported by the applicant, these cold tolerant translines grown in these field tests did not 

produce any pollen, the applicant has not observed any volunteer seedlings in or around the test 

sites where trees have flowered and produced seed capsules, and monthly field test monitoring 

observations have not identified any differences in diseases and insects or other non-target 

organisms between the transgenic and non-transgenic trees in the field test. Based on the 

analysis and information provided in the EA and supporting permit application, the new genes 

that are engineered into the trees should not pose any greater risk of loss of confinement and risk 

to the environment than those that are currently authorized by APHIS for confined field release. 

The addition of new genes to increase cold tolerance, alter lignin and growth should not 

compromise the ability of these to remain confined field tests. Adherence to the supplemental 

permit conditions established for the permit by the applicant will effectively limit any potential 

adverse impacts to the human environment. As discussed in Section V of the EA, APHIS 

considers the possibility of unintended exposure from moving GE Eucalyptus trees under the 

proposed action to be negligible to non-existent. 

Failure to Assess Cumulative Effects of all existing ArborGen permits 

One commenter expressed the concern that since there are now six active permits under which 

ArborGen is authorized by APHIS to grow GE Eucalyptus which include 32 unique locations 

within 7 States where up to a total of 654.7 acres of GE Eucalyptus could be planted, that the 

cumulative impacts of this proposal are substantial and significant and should be considered. 

Response: A cumulative impacts analysis is provided in Section V of the EA. As identified in 

the permit application, GE Eucalyptus trees with the new constructs would be released at 6 

locations. The new permit application does not expand the number of trees or total acreage that 

could be planted at any of the confined release sites currently approved by APHIS. As described 

in the EA and other EAs prepared by APHIS for authorizing ArborGen permits 

(http://www.aphis.usda.gov/brs/aphisdocs/06 32511lr ea.pdf and 

http://www.aphis.usda.gov/brs/aphisdocs/08 01410lr ea.pdf), the 32 field test sites are widely 

dispersed across the southeastern US. The sites are 900 miles apart from the farthermost east 

and west plantings. They average approximately 50 miles apart. The closest plantings have at 

least a mile separation between them, no individual planting is greater than 20 acres in size and 

there is no reason to assume that there will be any interaction between these distant sites. The 

small size of the field plots and the distance between test sites spread over thousands of miles 

indicate no cumulative effects would result from APHIS issuing the proposed field release 

permits. Therefore the only past, present, and reasonably foreseeable actions associated with the 

locations for the proposed releases under permit are those related to agricultural or forest tree 

production. Based on the analysis provided in the EA, APHIS has determined that there are no 

past, present, or reasonably foreseeable actions that would aggregate with effects of the proposed 

Page 24 of28 
action to create cumulative impacts or reduce the long-term productivity or sustainability of any 

of the resources (soil, water, ecosystem quality, biodiversity, etc.) associated with the release 

sites or the ecosystem in which they are situated. No resources will be significantly impacted 

due to cumulative impacts resulting from the proposed action. 

Failure to Consider a Reasonable Range of Alternatives 

One commenter said: "While the DEA contains only two alternatives, it actually only has one: 

the proposal contained in Arborgen's application. The other alternative is the "no action" 

alternative, which is to deny the application. It therefore fails to include a range of alternatives. 

The commenter suggests two additional alternatives should be analyzed in the DEA: 

• One which excludes the change in conditions, the allowed flowering, of genetically 

engineered Eucalyptus trees authorized under permits 09-070-1 01 rm and 11-041-1 01 rm, 

and 

• One which both excludes the change in conditions, the allowed flowering, of genetically 

engineered Eucalyptus trees authorized under permits 09-070-101rm and 11-041-101rm, 

but also does not allow the flowering of genetically engineered Eucalyptus trees at the 6 

new sites. 

Response: APHIS has prepared this EA in response to confined environmental release permit 

applications (APHIS Number 11-052-101rm) received from ArborGen LLC. This permit 

application was submitted in accordance with APHIS' regulations in 7 CFR part 340. In 

accordance with these regulations when APHIS receives an application for a permit for 

environmental release, the application is evaluated to determine whether the environmental 

release, with appropriate conditions imposed, can be carried out while preventing the 

dissemination and establishment of plant pests. The receipt of a permit application to introduce a 

genetically engineered organism requires a response from the Administrator: 

Administrative action on applications. After receipt and review by APHIS of the 

application and the data submitted pursuant to paragraph (a) of this section, including any 

additional information requested by APHIS, a permit shall be granted or denied (7 

CFR 340.4(e)). 

The applicant has provided the information associated with this request in the permit application 

and APHIS now must make a determination to either grant or deny the permit. Since APHIS 

decision is to either grant or deny the permit application, APHIS analysis of the two alternatives 

(No Action- Deny the permit and Preferred Alternative- Issue the APHIS Permit) identified in 

the EA are reasonable. The EA analyzes the potential cumulative effects of the proposed field 

test locations and has shown that the issuance of the permits will not have a significant impact on 

the human environment. Further analysis on a site by site basis or over a more restricted space 

or time will not provide any additional information or analysis that would be necessary to make 

an informed NEP A decision on the issuance of these permits. 

The alternatives suggested by the commenter have been analyzed in the EA as either the No 

Action Alternative or the Preferred Alternative. As clearly stated in Section V of the EA, GE 

Eucalyptus trees grown under the confined field release permits 09-070-101 rm and 11-041-

101 rm would not be allowed to flower under the No Action Alternative. An additional 

alternative that analyzes the potential impacts of not allowing GE Eucalyptus trees to flower at 

Page 25 of28 
any of the proposed 6 confined release sites would not provide any additional information to the 

decision maker than what is already provided under the Preferred Alternative. In effect, the 

supplemental permit conditions under the Preferred Alternative will have the same impact on the 

environment as would an alternative that does not allow the trees to flower. As described in the 

EA, the supplemental permit conditions (see Appendix VII of the EA) established for the permit 

will effectively limit the reproductive capabilities and establishment of this GE Eucalyptus 

outside the confined field trial locations. 

Failure to prepare an EA or EISon permits 09-070-lOlrm and 11-041-lOlrm 

A commenter said: "Because permits 09-070-101rm and 11-041-101rm and all previous permits 

were approved without completing either an EIS or an EA, there has not been an environmental 

assessment of any sort on the environmental effects of introducing Eucalyptus grandis X 

Eucalyptus urophylla. This includes, but is not limited to critical habitat analysis under ESA, 

effects on hydrology and effects of pollen on bee populations, as referenced in previous sections 

of this document. APHIS did not provide public notice or an opportunity for public comments or 

request an ESA determination from FWS. Therefore the environmental effects of all of these 

permits must be considered in any environmental analysis, immediate, cumulative or otherwise." 

Response: This comment is outside the scope of this EA. APHIS has prepared this EA 

specifically to evaluate and determine if there are any potentially significant impacts to the 

human environment in response to a confined environmental release and interstate movement 

permit application (11-052-101rm) received from ArborGen to authorize the planting of 

genetically engineered (GE) Eucalyptus hybrid clone (Eucalyptus grandis x Eucalyptus 

urophylla) to support proposed field research studies on six research sites in Alabama, Florida, 

Mississippi, and South Carolina. A discussion of potential impacts associated with permits 

authorized by APHIS for GE Eucalyptus is presented in the cumulative impacts analysis 

provided in Section V of the EA and the "cumulative effects" response above. 

All regulatory actions taken by APHIS, including the authorization of permits 09-070-101 rm and 

11-041-101rm, are conducted in accordance with: (1) The National Environmental Policy Act of 

1969 (NEPA), as amended (42 U.S.C. § 4321 et seq.), (2) regulations ofthe Council on 

Environmental Quality for implementing the procedural provisions ofNEPA ( 40 CFR parts 

1500-1508), (3) USDA regulations implementing NEPA (7 CFR part 1b), and (4) APHIS' NEPA 

Implementing Procedures (7 CFR part 372). 

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