Monsanto Petition (11-188-01p) for Determination of 

Non-regulated Status of MON 88302 Canola

OECD Unique Identifier: 

MON 883Ø2-9

Plant Pest Risk Assessment 

May 2013

Agency Contact 

Cindy Eck

Biotechnology Regulatory Services

4700 River Road

USDA, APHIS 

Riverdale, MD 20737 

Fax: (301) 734-8669

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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. 
TABLE OF CONTENTS

A. Introduction................................................................................................................. 1

B. Development of [GE Crop Event] (includes description of the genetic 

modification)........................................................................................................... 3

C. Expression of the Gene Product and Changes to Plant Metabolism........................... 6

D. Potential Impacts on Disease and Pest Susceptibilities.............................................. 9

E. Potential Impacts on Nontarget Organisms Beneficial to Agriculture ..................... 12

F. Potential for Enhanced Weediness of [GE Crop Event]........................................... 13

G. Potential Impacts on the Weediness of Any Other Plants with which It Can 

Interbreed .............................................................................................................. 17

H. Potential Changes to Agriculture or Cultivation Practices....................................... 21

I. Potential Impacts from Transfer of Genetic Information to Organism with 

which [GE Crop Event] Cannot Interbreed .......................................................... 23

J. Conclusion ................................................................................................................ 24

K. References................................................................................................................. 25

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A. Introduction

Monsanto Company (referred hereafter as Monsanto) has petitioned the Animal and Plant 

Health Inspection Service (APHIS) of the United States Department of Agriculture 

(USDA) for a determination that the genetically engineered (GE) glyphosate herbicideresistant1 canola event MON 88302 (referred hereafter as MON 88302) is unlikely to 

pose a plant pest risk and, therefore, should no longer be a regulated article under the 

APHIS’ 7 Code of Federal Regulations (CFR) part 340. This petition was assigned the 

number 11-188-01p, and is hereafter referenced as Monsanto 2011. APHIS administers 7 

CFR part 340 under the authority of the plant pest provisions of the Plant Protection Act 

(PPA) of 2000 (7 U.S.C. 7701 et seq.)

2

. This plant pest risk assessment was conducted to 

determine if MON 88302 is unlikely to pose a plant pest risk.

APHIS regulations in 7 CFR part 340 regulate the introduction (importation, interstate 

movement, or release into the environment) of certain GE organisms and products. A GE 

organism is no longer subject to the plant pest provisions of the PPA or to the regulatory 

requirements of Part 340 when APHIS determines that it is unlikely to pose a plant pest 

risk. A GE organism is considered a regulated article under Part 340 if the donor 

organism, recipient organism, or vector, or vector agent used in engineering the organism 

belongs to any genera or taxa designated in 7 CFR 340.2 and is also considered a plant 

pest. A GE organism is also regulated under 7 CFR part 340 when APHIS has reason to 

believe that the GE organism may be a plant pest or APHIS does not have sufficient 

information to determine if the GE organism is unlikely to pose a plant pest risk. MON 

88302 was produced by transformation of canola tissue using Agrobacterium 

tumefaciens, a plant pest, and some of the sequences (i.e., enhancer sequence from 35S 

promoter from figwort mosaic virus, codon-optimized aroA gene sequence from 

Agrobacterium sp. strain CP4., and right and left T-DNA border sequences from A. 

tumefaciens) used in the transformation process (Monsanto 2011) are also from plant 

pests organisms listed in 7 CFR 340.2. Monsanto has conducted introductions of MON 

88302 as a regulated article under APHIS-authorized notifications since 2005 (Appendix 

Table A-1, p. 218, Monsanto 2011), in part, to gather information to support that it is 

unlikely to pose a plant pest risk. 

Potential impacts in this Plant Pest Risk Assessment are those that pertain to plant pest 

risk characteristics associated with MON 88302 and its progeny and their use in the 

1

Monsanto has described the phenotype of MON 88302 as “herbicide tolerant” and historically APHIS has also referred to GE plants 

with reduced herbicide sensitivity as herbicide tolerant. However, the phenotype would fall under the Weed Science Society of 

America’s definition of “herbicide resistance” since MON 88302 has an “inherited ability to survive and reproduce following 

exposure to a dose of herbicide normally lethal to the wild type” (WSSA 1998). By the WSSA definition, “resistance [to an herbicide] 

may be naturally occurring or induced by such techniques as genetic engineering or selection of variants produced by tissue culture or 

mutagenesis. ”Herbicide tolerance, by the WSSA definition, only applies to plant species with an “inherent ability to survive and 

reproduce after herbicide treatment. This implies that there was no selection or genetic manipulation to make the plant tolerant; it is 

naturally tolerant."

2

Plant Protection Act in 7 U.S.C. 7702 § 403(14) defines plant pest as: “Plant Pest - The term “plant pest” means any living stage of 

any of the following that can directly or indirectly injure, cause damage to, or cause disease in any plant or plant product: (A) A 

protozoan. (B) A nonhuman animal. (C) A parasitic plant. (D) A bacterium. (E) A fungus. (F) A virus or viroid. (G) An infectious 

agent or other pathogen. (H) Any article similar to or allied with any of the articles specified in the preceding subparagraphs.” 

1

 
absence of confinement. APHIS utilizes data and information submitted by the applicant, 

in addition to current literature, to determine if MON 88302 is unlikely to pose a plant 

pest risk. APHIS regulations in 7 CFR 340.6(c) specify the information needed for 

consideration in a petition for nonregulated status. APHIS will evaluate information 

submitted by the applicant related to plant pest risk characteristics, expression of the gene 

product, new enzymes, or changes to plant metabolism, potential impacts of genetic 

modifications on disease and pest susceptibilities, effects of the regulated article on 

nontarget organisms, weediness of the regulated article, impact on the weediness of any 

other plant with which it can interbreed, changes to agricultural or cultivation practices 

that may impact diseases and pests of plants, indirect plant pest effects on other 

agricultural products, and transfer of genetic information to organisms with which it 

cannot interbreed. 

APHIS may also consider information relevant to reviews conducted by other agencies 

that are part of the ‘Coordinated Framework for the Regulation of Biotechnology’ (51 FR 

23302, June 26, 1986). Under the Coordinated Framework, the oversight of 

biotechnology-derived plants rests with the APHIS, the Food and Drug Administration 

(FDA), and the Office of Pesticide Programs of the U.S. Environmental Protection 

Agency (EPA). Depending on its characteristics, certain biotechnology-derived products 

are subjected to review by one or more of these agencies. 

The EPA under the Federal Insecticide, Fungicide and Rodenticide Act (FIFRA), 

regulates the distribution, sale, use and testing of pesticidal substances produced in plants 

and microbes, including those pesticides that are produced by an organism through 

techniques of modern biotechnology. The EPA also sets tolerance limits for residues of 

pesticides on and in food and animal feed, or establishes an exemption from the 

requirement for a tolerance, under the Federal Food, Drug and Cosmetic Act (FFDCA). 

The post-emergence (in-crop) use of glyphosate in previously deregulated Roundup 

Ready canola RT73 was first approved by the EPA in March 1999 (Monsanto 2011). 

Monsanto has submitted a request for amended labeling to the EPA in February 2011 for 

EPA Registration Numbers 524-537 (Roundup WeatherMAX® Herbicide) and 524-549 

(Roundup PowerMAX® Herbicide) that proposes to modify the current use pattern of 

glyphosate in canola based on MON 88302. The amended labeling request pertains to 

use of glyphosate during an expanded window of application and at rates higher than 

those currently recommended and authorized (p. 13, Monsanto 2011). EPA is currently 

reviewing the label changes to add the increased rate and later application of glyphosate 

to commercial herbicide-resistant canola production. EPA’s assessment will analyze 

risks to non-target organisms to determine if the label is sufficient to meet EPA’s 

standards for registration; “reasonable certainty of no harm to humans” and “no 

unreasonable adverse effects on the environment.” If these standards are not met, EPA 

will apply appropriate risk mitigation strategies and propose label modifications to 

address the specific concerns. After EPA has completed it assessments and provided 

these to APHIS, APHIS will update this PPRA as appropriate.

2
The FDA under the FFDCA is responsible for ensuring the safety and proper labeling of 

all plant-derived foods and feeds, including those developed through modern 

biotechnology. To help sponsors of foods and feeds derived from genetically engineered 

crops comply with their obligations, the FDA encourages them to participate in its 

voluntary consultation process. 

In compliance with this policy, Monsanto initiated a consultation with the FDA (FDA 

BNF No. 127) on the food and feed safety and compositional assessment of MON 88302

(p. 28, Monsanto 2011). Monsanto submitted a safety and nutritional assessment 

summary document to the FDA in March 2011 and received a completed consultation 

letter from the FDA in April 2012. A copy of the text of this letter responding to BNF 

127, as well as a copy of the text of FDA’s memorandum summarizing the information in 

BNF 127, is available for public review via the FDA Completed Consultations on 

Bioengineered Foods page at www.fda.gov/bioconinventory. 

B. Development of MON 88302 Canola

Canola (Brassica spp) is an oil seed crop primarily cultivated in China, India, Europe, 

and Canada and is becoming popular in the United States and Australia (OECD 2012).

There are two main types of B. napus: 1) oil-yielding oleiferous rape, of which one subset 

with specific quality characteristics is often referred to as "canola" (vernacular name), 

and 2) vegetable and/or forage type: the tuber-bearing swede or rutabaga (B. napus var. 

napobrassica, rutabaga or Swede) and Siberian or rape kale (B. napus var. 

pabularia)(OECD 2012). At present, three species of Brassica (B. napus, B. rapa, and B. 

juncea) have commercialized varieties with "double low" characteristics, i.e. low erucic 

acid content (<2%) in the fatty acid profile and very low glucosinolate content (<30 

micromoles/g) in the air-dried oil-free meal, characteristics desirable for high-quality 

vegetable oil and high-quality animal feed, respectively (CCC 2003). In North America 

these species are considered to be of "canola" quality. There are spring (annual) and 

winter (biennial) biotypes of canola varieties. Spring canola is planted in early spring 

and harvested in late summer. It is a cool season crop that is grown in western Canada, 

southern Australia, northern China, and in the northern Great Plains region of the U.S. 

Winter canola is planted in the fall, requires exposure to winter cold (vernalization) to 

flower, and is grown in parts of northern Europe, central China, the northwestern U.S. 

and in the central portions of the U.S. Great Plains (OECD 1997, 2012). The spring 

biotypes are typically lower yielding than the winter types, but require considerably less 

time to complete their life cycle (OECD 1997). Spring canola varieties are available in 

all three Brassica species while winter canola varieties are created only in B. napus and 

B. rapa species (CCC 2003).

In the U.S. canola is grown in three geographical regions – the Northern and Southern 

Great Plains and the Pacific Northwest (Figure VIII-1, p. 131 and Table VIII-2, p. 132,

Monsanto 2011). For the growing season 2010, the largest U.S. canola producing state 

was North Dakota (1,280,000 acres or ~ 88%) followed by Oklahoma with 60,000 acres

(~ 4%) (Table VIII-2, p. 132, Monsanto 2011). Nearly all the canola grown in North 

Dakota is the B. napus spring canola type (NDSU 2005) while Oklahoma primarily 

grows the B. napus winter canola type (Monsanto 2011, p.129).

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MON 88302, a spring canola biotype (B. napus), has been genetically modified to 

express 5-enolpyruvylshikimate-3-phosphate synthase (termed CP4 EPSPS) enzyme. 

The CP4 EPSPS enzyme, encoded by the cp4 epsps gene, confers resistance to 

glyphosate-containing herbicides by the production of CP4 EPSPS protein that is less 

sensitive to inhibition by glyphosate compared to the endogenous plant EPSPS (Padgette 

et al. 1996; Funke et al. 2006). As detailed later in this document, MON 88302 utilizes 

an improved promoter sequence to enhance CP4 EPSPS expression in male reproductive 

tissues to facilitate glyphosate herbicide application at later stages of plant development 

without harming male reproductive tissues. According to Monsanto, such late 

application of glyphosate herbicide is not possible with currently available glyphosate 

herbicide-resistant canola varieties, and later application has the advantage of providing 

greater flexibility to growers to manage weeds in canola fields (p. 4, Monsanto 2011).

Description of the genetic modifications

As described in the petition (p. 33, Monsanto 2011) MON 88302 was developed through 

Agrobacterium-mediated transformation of hypocotyls (Radke et al. 1992) of a donor 

spring canola variety, Ebony, utilizing a binary plasmid vector PV-BNHT2672. The PVBNHT2672 plasmid vector contained two parts as summarized below: 1) a Transfer DNA 

(T-DNA) sequence containing the cp4 epsps plant expression cassette, and 2) backbone 

sequences necessary for maintenance or selection of the plasmid vector in bacteria, but 

which are not expected to be transferred to the plant (Figure III-1, p. 35 & Table III-1, pp.

38-39, Monsanto 2011). 

T-DNA cp4 epsps cassette

• Right Border (RB) Sequence: A specific DNA region from A. tumefaciens used 

for T-DNA transfer (Depicker et al. 1982; Zambryski et al. 1982).

• FMV/Tsf1 Promoter: Chimeric promoter consisting of the promoter of the Tsf1

(Twin Sister of Flowering Locus 1) gene from the Arabidopsis thaliana encoding 

elongation factor EF-1α (Axelos et al., 1989) and enhancer sequences from the 

35S promoter from the FMV (Figwort Mosaic Virus) (Richins et al. 1987). 

• Tsf1 Leader Sequence: 5' untranslated leader (exon 1) from the A. thaliana Tsf1

gene encoding elongation factor EF-1α (Axelos et al. 1989).

• Tsf1 intron: Intron from the A. thaliana Tsf1 gene encoding elongation factor EF1α (Axelos et al. 1989).

• CTP2 targeting sequence: Targeting sequence from the shkG gene encoding the 

chloroplast transit peptide region of A. thaliana EPSPS (Herrmann 1995; Klee et 

al. 1987) that directs transport of the CP4 EPSPS protein to the chloroplast.

• cp4 epsps coding sequence: Codon optimized coding sequence of the aroA gene 

from the Agrobacterium sp. strain CP4 encoding the CP4 EPSPS protein (Barry et 

al. 2001; Padgette et al. 1996).

• rbcS2-E9 gene terminator: 3' untranslated sequence from the rbcS2 gene of Pisum

sativum (pea) encoding the Rubisco small subunit (Coruzzi et al. 1984).

• Left Border (LB) sequence: A specific DNA region from A. tumefaciens used for 

T-DNA transfer (Barker et al. 1983; Zambryski et al. 1982).

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Backbone sequence

• Origin of replication (oriV): Origin of replication from the broad host range 

plasmid RK2 for maintenance of plasmid in Agrobacterium (Stalker et al. 1981).

• Repressor of primer (rop) protein coding sequence: Coding sequence for repressor 

of primer protein for maintenance of plasmid copy number in E. coli (Giza and 

Huang 1989).

• Origin of replication (ori-pBR322): Origin of replication from pBR322 for 

maintenance of plasmid in E. coli (Sutcliffe 1979).

• Promoter-coding sequence-Terminator sequence for aadA gene: Bacterial 

promoter, coding sequence, and 3′ untranslated region for an aminoglycosidemodifying enzyme,

• 3''(9)-O-nucleotidyl-transferase from the transposon Tn7 (Fling et al. 1985) that 

confers

• Spectinomycin and streptomycin antibiotic resistance.

• In addition to the above sequences, both the cp4 epsps cassette and the backbone 

contain four or five intervening sequences of various lengths to facilitate DNA 

cloning. 

The binary plasmid vector PV-BNHT2672 is disarmed. Disarmed binary plasmid vectors 

lack T-DNA sequences from Ti (Tumor-inducing) – plasmids normally responsible for 

the formation of crown gall tumors upon A. tumefaciens infection (Hoekema et al. 1983; 

Hellens et al. 2000). Furthermore, following transformation the plant tissue was treated 

with antibiotics carbenicillin, ticarcillin disodium and clavulanate potassium to inhibit the 

growth of excess Agrobacterium (Hellens et al. 2000; p. 33, Monsanto 2011). Plant 

pathogen sequences inserted into MON 88302, i.e. the T-DNA border sequences from 

Agrobacterium and the enhancer sequences from FMV do not cause plant disease.

Monsanto provided evidence demonstrating that,

• the DNA inserted into the MON 88302 canola genome is present at a single locus, 

and contains one functional copy of cp4 epsps gene expression cassette with 

truncated portions of the T-DNA right border (42 bp) and left border (273 bp)

(Table IV-2, p. 46 and section IV.A., pp. 47-50, Monsanto 2011); 

• the final product does not contain any of the backbone sequences from the 

plasmid PV- BNHT2672 outside of the T-DNA region or cp4 epsps expression 

cassette borders as determined through Southern blot analysis (section IV-B., pp. 

51-55, Monsanto 2011);

• the T-DNA sequence in MON 88302 (Table IV-2, p.46, Monsanto 2011) is 

identical to the corresponding T-DNA sequence of the original donor plasmid PVBNHT2672 (as confirmed by DNA sequence analysis) (section IV.C., pp. 56-57, 

Monsanto 2011);

• the inserted cp4 epsps expression cassette DNA was stably inherited across four 

breeding generations (section IV.E., pp. 60-62, Monsanto 2011); and,

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• the cp4 epsps expression cassette in MON 88302 resides at a single locus within 

the canola genome and is inherited according to Mendelian principles of 

inheritance (as determined by Chi-square analysis for three segregating 

populations) (section IV-F., pp. 63-66, Monsanto 2011).

Insertion of foreign genetic materials tends to induce mutations at sites of insertion 

(generally referred to as insertional mutations) in recipient genomes (Nacry et al. 1998; 

Laufs et al. 1999). Monsanto examined the T-DNA insertion site in MON 88302 and 

corresponding conventional control line using Polymerase Chain Reaction (PCR) and 

sequence analyses (Appendix B, pp. 223-224, Monsanto 2011) and discovered that MON 

88302 contained transformation induced mutations at the flanking site adjacent to the 3’ 

end of the T-DNA insert (section IV.D., pp. 58-59, Monsanto 2011). There was a 9 base 

pair insertion, a 29 base pair deletion, and a single nucleotide difference between the 

conventional control sequence and the genomic DNA sequence flanking the 3' end of the 

T-DNA insert in MON 88302. According to Monsanto, the observed insertion-deletion 

mutation (indel mutation) presumably resulted from double-stranded break repair 

mechanisms in the plant during the Agrobacterium-mediated transformation process 

(Salomon and Puchta 1998) while a single nucleotide difference was most likely caused 

by a single nucleotide polymorphism (SNP) segregating in the canola population (Trick 

et al. 2009). As discussed later in this document, none of these mutations altered the 

function of cp4 epsps gene or exhibited deleterious phenotypes in MON 88302.

C. Expression of the Gene Product, Enzymes or Changes to Plant 

Metabolism

USDA-APHIS assessed whether changes in plant metabolism or composition in MON 

88302 canola are likely to alter its plant pest risk. The assessment encompasses a 

consideration of the expressed protein or enzyme and its effect on plant metabolism and 

an evaluation of whether the nutrients and anti-nutrient levels in harvested seed derived 

from MON 88302 are comparable to those in the conventional canola control variety 

Ebony or to other reference canola cultivars considered for the composition analysis. 

Host plant quality (including such components as carbon, nitrogen, amino acid sources, 

trace elements, and defensive metabolites) is known to affect herbivore performance and 

fecundity; and higher-trophic level interactions, such as the performance of predators and 

parasitoids, may also be affected (reviewed by Awmack and Leather 2002). Similarly a 

vast array of secondary metabolites in plants is known to provide defense against 

microbes (Dixon, 2001). Thus APHIS assessed whether changes in host plant quality 

could have the potential to affect MON 88302 canola’s performance against pest and 

disease incidences.

The enzyme, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), catalyzes one of

the enzymatic steps of the shikimic acid pathway, and is the target for the broad spectrum

herbicide glyphosate, the active ingredient in Roundup agricultural herbicides (Haslam

1993; Herrmann and Weaver 1999; Steinrücken and Amrhein 1980). Glyphosate -

herbicide resistance in MON 88302 comes from the expression of a bacterial 

(Agrobacterium sp. strain CP4) gene cp4 epsps. The cp4 epsps gene encodes CP4 EPSPS 

protein, which like plant EPSPS protein, catalyzes the transfer of the enolpyruvyl group 

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from phosphoenolpyruvate (PEP) to the 5-hydroxyl of shikimate-3-phosphate (S3P), 

thereby yielding inorganic phosphate and 5-enolpyruvylshikimate-3-phosphate (EPSP) 

(Alibhai and Stallings 2001). Shikimic acid is a substrate for the biosynthesis of the 

aromatic amino acids (phenylalanine, tryptophan and tyrosine) and other aromatic 

molecules necessary for plant growth. In conventional plants, glyphosate blocks the 

biosynthesis of EPSP thereby depriving plants of essential amino acids (Steinrücken and 

Amrhein 1980; Haslam 1993). The CP4 EPSPS protein is similar and functionally 

identical to endogenous plant EPSPS enzymes, but has a much reduced affinity for 

glyphosate relative to endogenous plant EPSPS (Padgette et al. 1996). Therefore, in 

glyphosate herbicide-resistant plants, requirements for aromatic amino acids and other 

metabolites are met by the continued action of the CP4 EPSPS enzyme in the presence of 

glyphosate (Padgette et al. 1996).

In MON 88302 the cp4 epsps coding sequence encodes a 47.6 kDa EPSPS protein 

consisting of a single polypeptide of 455 amino acids (Figure III-3, p. 40, Monsanto 

2011; Padgette et al. 1996). Because CP4 EPSPS protein has a long history of use in 

various genetically engineered (GE) crops to provide herbicide-resistance, its 

physiochemical and functional properties along with food and/or feed safety data were 

previously examined on an identical version of CP4 EPSPS protein produced in E. coli

(Harrison et al. 1996). Thus E. coli produced CP4 EPSPS protein has become an industry 

standard for comparative studies. A comparison of the MON 88302-produced CP4 

EPSPS protein to the E. coli-produced CP4 EPSPS protein confirmed the structural and 

functional identity of the MON 88302-produced CP4 EPSPS protein with that of E. coli

produced CP4 EPSPS protein (Appendix C, pp. 226-244, Monsanto 2011). Furthermore, 

since CP4 EPSPS proteins isolated from other Roundup Ready crops have been 

previously demonstrated to be equivalent to the E. coli-produced CP4 EPSPS protein, 

then according to Monsanto by inference the MON 88302-produced CP4 EPSPS protein 

must be equivalent to the CP4 EPSPS proteins expressed in other Roundup Ready crops 

that have been deregulated by USDA-APHIS (Section V, pp. 68-70, Monsanto 2011).

Monsanto estimated CP4 EPSPS protein levels in eight tissue types covering forage, 

seed, leaf and root (Appendix D, pp. 245-249, Monsanto 2011) collected from three 

experimental fields in the U. S. (Idaho, Minnesota and North Dakota) and three in 

Canada (two sites in Manitoba and one in Saskatchewan) during the 2009 growing 

season. According to Monsanto, the six selected field sites were representative of canola 

producing regions suitable for commercial production in the U.S. and Canada. Monsanto 

observed MON 88302 CP4 EPSPS protein expression in all the various tissue types: its 

level was highest in leaf (180 - 230 μg/g dry weight (dw)) followed by forage, root and 

seed (27 μg/g dw) (Table V-1, p. 71, Monsanto 2011). The CP4 EPSPS protein 

expression levels in leaf (forage) and seed tissue in MON 88302 reported on a fresh 

weight basis (Table V-1, p. 71, Monsanto 2011) are within the range reported for the 

previously deregulated glyphosate resistant canola RT73 (Table 6, p. 36, Monsanto 

1998). Although data on CP4 EPSPS protein in pollen were not reported in the petition, 

other studies suggest that it is more highly expressed in MON 88302 compared to RT73. 

Feng et al. (2010) describes the difference in sensitivity to glyphosate between RT73 and 

MON 88302, the ‘second generation RR2’ GR canola. Pollen viability drops to zero 

percent when RT73 is sprayed at 4-10-leaf stages with 1.33 times the maximum single 

7
application rate of 0.6 kg acid equivalents (a.e.)/hectare ha, whereas no reduction in 

pollen viability is seen in MON 88302 pollen even with 6 times this application rate. Also 

at this 6 x rate, MON 88302 is able to produce approximately 1.5 times more average 

seed weight per plant than RT73 (Figure 3.2 in Feng et al. 2010). MON 88302 also 

suffers no leaf chlorosis when sprayed with an 8 x rate of glyphosate. 

Monsanto carried out a compositional assessment of seed samples obtained from MON 

88302, control, and seven commercial reference varieties (Appendix Table E-1, p. 250, 

Monsanto 2011) using the principles and analytes outlined in the OECD consensus 

document for canola composition (OECD 2001) to assess whether levels of key nutrients, 

toxicants and anti-nutrients in MON 88302 were equivalent to levels in the conventional 

control and to the composition of commercial reference varieties. The OECD consensus 

document does not recommend analysis of canola forage, as canola forage is rarely 

consumed by animals and is not a source of nutrition for humans. The seven different 

commercial reference varieties were included across all sites of the field production to 

provide data on natural variability of each compositional component analyzed. The 

samples for compositional assessment were collected in the 2009 growing season from 

two U.S. sites (Minnesota and North Dakota) and three Canadian sites (two sites in 

Manitoba and one site in Saskatchewan). The five field sites were chosen to represent the 

typical canola growing regions of the United States and Canada. In addition to the 

conventional weed control programs, MON 88302 plots were treated at the 5-6 leaf stage 

with a glyphosate application at a target rate of 1.6 lb a.e. per acre (1800 g a.e./ha) (p. 77, 

Monsanto 2011).

Nutrients assessed included proximates (ash, carbohydrates, moisture, protein, and total 

fat), fiber, amino acids, fatty acids, vitamin E, and minerals. The toxicants assessed in 

seed included erucic acid and glucosinolates. The anti-nutrients assessed in seed 

included phytic acid and sinapine (see Appendix E for analytical methods. pp. 250-259, 

Monsanto 2011). In all, according to Monsanto, 70 different components were measured. 

Of those 70 components, 18 nutrients and one toxicant (18 fatty acids, including erucic 

acid, and one mineral) had more than 50% of the observations below the assay limit of 

quantitation (LOQ), and therefore were excluded from the statistical analyses. 

Monsanto employed six statistical comparisons on the MON 88302 compositional data. 

One comparison was based on compositional data combined across all five field sites 

(combined-site analysis) and five separate comparisons were conducted on data from 

each of the individual field sites. Data from the commercial reference varieties were 

combined across all sites and used to calculate a 99% tolerance interval for each 

compositional component to define the natural variability of each component in canola 

varieties that have a history of safe consumption, and that were grown concurrently with 

MON 88302 and the conventional control in the same trial (see p. 78, Monsanto 2011 for 

further details about data interpretations). 

No statistically significant differences were observed between MON 88302 canola and 

the control canola mean values for a majority of the analyzed nutrients except for total 

dietary fiber (TDF) and seven fatty acids (16:1 palmitoleic acid, 18:0 stearic acid, 18:1 

oleic acid, 18:2 linoleic acid, 18:3 linolenic acid, 20:0 arachidic acid, and 22:0 behenic 

8
acid) (Tables VI-1 and VI-2, pp. 83-100, Monsanto 2011). However, mean values for all 

of the nutrient components found to be significantly different (α = 0.05) from the 

combined-site analysis of MON 88302 were within the 99% tolerance interval 

established from the commercial reference varieties grown concurrently (Tables VI-1 and 

VI-2, pp. 83-100, Monsanto 2011); and with the exception of TDF, for which no 

commercial reference values have been published, all of the compositional components 

identified as significantly different from the conventional control were also within the 

natural variability of these components in commercial canola composition as published in 

the scientific literature (Table VI-4, p. 102-103, Monsanto 2011). 

Similar to nutrient analysis, MON 88302 and control were similar in anti-nutrient 

composition except for alkyl glucosinolate, which was slightly reduced in MON 88302 

(Tables VI-1 to VI-3, pp. 83-101, Monsanto 2011). Although in the combined-site 

analysis a significant difference in alkyl glucosinolates was observed for MON 88302; 

the mean value was lower than the conventional mean value, and the difference was not 

consistently observed at the individual sites. The combined-site mean alkyl 

glucosinolates value for MON 88302 was within the natural variability of commercial 

canola defined by the 99% tolerance interval established from the concurrently grown 

commercial reference varieties that have a history of safe consumption, and the value was 

within the safety threshold for canola (pp. 81-82, Monsanto 2011). Higher 

glucosinolinates in some Brassica species have also been reported to reduce the feeding 

rates of larvae of the pest species White cabbage butterflies (Pieris rapae) and the 

fecundity of the Cabbage aphid (Brevicoryne brassicae) (Awmack and Leather 2002). 

However, as noted in the next section of this PPRA, pest damage ratings for cabbage 

worms (Pieridae) and aphids were not different between the 88302 canola and control 

canola (Table G7, p. 331, Monsanto 2011, nor was there any significant difference in 

aphid abundance (Table G-9, p. 333, Monsanto 2011). 

Results of the comparison indicate that the composition of the seed of MON 88302 is 

similar to that of the conventional control and within the natural variability of 

commercial reference varieties. These findings support the conclusion that nutrients in 

seed from MON 88302 are compositionally equivalent to those in conventional canola 

varieties with a history of safe usage.

Previously CP4 EPSPS protein was part of 13 deregulated crop events, including canola 

(Petitions for Nonregulated Status Granted or Pending by APHIS, 

http://www.aphis.usda.gov/biotechnology/not_reg.html; Information Systems for 

Biotechnology, http://www.isb.vt.edu/search-petition-data.aspx). So far there are no 

reports of plant pest characteristics exhibited by them. Based on all the above noted 

considerations, APHIS concludes that MON 88302 poses no more of a plant pest risk 

from new gene products, changes to plant metabolism or composition than conventional 

canola varieties.

D. Potential Impacts on Disease and Pest Susceptibilities

USDA-APHIS assessed whether MON 88302 is likely to have significantly increased 

disease and pest susceptibility because of the introduced cp4 epsps gene compared to the 

9
control canola variety. This assessment encompasses a thorough consideration of 

introduced traits, their impact on agronomic traits (discussed later in the document) and 

plant composition (discussed earlier), and quantitative and/or qualitative data on pest and 

disease responses. Important changes are those which would (1) affect not only the new 

GE crop, but that would also result in significant introduction or spread of a damaging 

pest or disease to other plants; and/or (2) result in the introduction, spread, and/or 

creation of a new disease or (3) result in a significant exacerbation of a pest or disease for 

which APHIS has a control program. Any increase in pest or disease susceptibility 

should be evaluated with respect to the context of currently cultivated varieties, the 

ability to manage the pest or disease, and the potential impact on agriculture.

Plant Protection and Quarantine (PPQ) is an APHIS program that safeguards agriculture 

and natural resources from the entry, establishment, and spread of animal and plant pests 

and noxious weeds into the United States of America; and supports trade and exports of 

U.S. agricultural products. PPQ responds to many new introductions of plant pests to 

eradicate, suppress, or contain them through various programs in cooperation with state 

departments of agriculture and other government agencies. These may be emergency or 

longer term domestic programs that target a specific pest. A variety of insect, plant 

disease, mollusk, nematode, or weed programs exist (see http://www.aphis.usda.gov/ 

plant_health/plant_pest_info/index.shtml ); however none specifically target pests of 

canola.

The complex of insects that feed upon the Brassicas is one of the important factors 

limiting the production of commercial Brassica crops (Lamb 1989: Weiss et al. 2009). 

Brassicaceous plants produce a family of sulphur compounds called glucosinolates whose 

breakdown products are attractants and stimuli for feeding and oviposition but, on the 

other hand, act as deterrents or toxins for herbivores not adapted to plants of the 

Brassicaceae. Some of the more important insect pests of canola are listed in Table VIII4 (p. 148, Monsanto 2011). Likewise, Brassica crops are subject to a broad range of 

pathogens (Kharbanda et al. 2001). Out of all the diseases affecting Brassica crops, the 

three most troublesome diseases are blackleg or stem canker (Leptosphaeria maculans); 

Sclerotinia stem rot (Sclerotinia sclerotiorum); and clubroot (Plasmodiophora brassicae).

As detailed in the petition (p. 116, Monsanto 2011), in the 2009 growing season, 17 trial

locations (8 in the U.S. and 9 in Canada) (Table VII-3, p. 118, Monsanto 2011) were 

selected in order to provide agronomic and pest and disease observations representative 

of the major spring annual canola growing regions in the U.S. and Canada, where 

commercial canola production and sales of MON 88302 canola are expected. Agronomic 

practices used to prepare and maintain each field site were characteristic of each 

respective region. Mon 88302 and the conventional control were planted at all field sites, 

while 24 commercial reference varieties included in the field trials (see Appendix Table 

G-1, p. 314, Monsanto 2011) were site-specific, locally-adapted genotypes, including a 

few glyphosate or glufosinate herbicide-resistant varieties. In addition, insect pests and 

diseases at the Canada locations are typical of those found in the U.S. All plots of MON 

88302, the conventional control, and the commercial reference varieties at each site were 

uniformly managed in order to assess whether the introduction of the glyphosatetolerance trait altered the phenotypic and agronomic characteristics or the environmental 

10
interactions (biotic and abiotic stress) of MON 88302 compared to the conventional 

control. Glyphosate herbicide was not applied to any of the plots in the trial. The same 

observations and data collection were not made for every site. Although some data were 

missing or excluded from statistical analyses or observational interpretations for a variety 

of technical and logistic reasons (Appendix G, Table G3, pp. 321-322, Monsanto 2011), 

the sample size included for statistical analyses on each variable was large enough to 

perform the necessary statistical analyses.

Monsanto conducted three types of pest and disease analyses—(1) qualitative assessment 

at all 17 sites for disease damage (Appendix Table G-6, p. 330, Monsanto 2011) and 

arthropod related damage (Appendix Table G-7, p. 331. Monsanto 2011); (ii) quantitative 

assessment at four sites for flea beetle (Chrysomelidae) and seedpod weevil 

(Ceutorhynchus obstrictus) damage (Appendix Table G-8, p. 332, Monsanto 2011); and, 

(iii) abundance of pest arthropods at four sites (Appendix Table G-9, p. 333-334, 

Monsanto 2011). A detailed description about the data collection sites, techniques, and 

statistical analyses has been provided in Appendix G (pp. 312-338, Monsanto 2011). 

In a qualitative assessment of plant response to disease damage and arthropod damage, no 

differences were observed between MON 88302 and the conventional control for any of 

the 141 comparisons involving 16 assessed diseases or for any of the 165 comparisons for 

any of the 13 assessed arthropods among all observations at the sites (Table VII-5, p. 

123; Appendix Tables G-6 and G-7, pp. 330-331, Monsanto 2011). Although a minor yet 

statistically significant difference was detected in flea beetle damage at one site for one 

observation (Table G-8, p. 332, Monsanto 2011), in the combined-site analysis for a 

quantitative assessment of flea beetle (Chrysomelidae) damage and seedpod weevil 

damage, no statistically significant differences were detected between MON 88302 and 

the conventional control from the four sites evaluated (Table VII-6, p. 124, Monsanto 

2011). Furthermore, pest and beneficial arthropods (the discussion on ‘beneficial 

arthropods’ is presented in the following section “Potential Effects on Nontarget 

Organisms...”) were collected at the four experimental locations four times during the 

2009 growing season (Section G.8, p. 337, Monsanto 2011 for sampling strategy and 

statistical analyses). According to Monsanto, the arthropods assessed often varied 

between collections from a site and between sites due to differences in temporal activity 

and geographical distribution of the taxa (Section G.8, p. 317, Monsanto 2011). Pest 

arthropod abundance data did not reveal any consistent trends that would suggest an 

increase in pest abundance for MON 88302 compared to the control variety for aphids 

(Aphididae), Bertha armyworms (Mamestra configurata), diamondback moth larvae 

(Plutella xylostella), flea beetles (Chrysomelidae), lygus bugs (Miridae), or thrips 

(Thripidae) (Table G-9, pp.333-334, Monsanto 2011) and no significant difference was 

detected for any of the 36 observations carried out across the four experimental sites 

(Table VII-5, p. 123, Monsanto 2011).

As discussed earlier there were no significant changes in MON 88302 compositions that 

would render MON 88302 more susceptible to pests and diseases over its control or 

reference canola varieties. As presented later in this document, the observed agronomic 

traits also did not reveal any significant changes that would indirectly indicate that MON 

88302 is or could be relatively more susceptible to pests and diseases over control or 

11
reference canola varieties. Thus MON 88302 is expected to be susceptible to the same 

plant pathogens and insect pests as conventional canola. The introduced genes did not 

significantly alter the observed insect pest infestation and disease occurrence or resulting 

damage on MON 88302 over the control line. For this reason, there is also unlikely to be 

any indirect plant pest effects on other agricultural products.

E. Potential Impacts on Nontarget Organisms, Including Those 

Beneficial to Agriculture

MON 88302 is not engineered for pest resistance, thus there are no ‘target’ species, and 

thus no ‘nontarget’ species either. However, APHIS assessed whether exposure or 

consumption of herbicide-resistant MON 88302 containing the CP4 EPSPS protein 

would have an adverse effect on beneficial species or wildlife associated with canola. 

Monsanto provided the following information justifying the safety of MON 88302 canola 

(Section V.D., pp. 72-75, Monsanto 2011):

(i) The donor organism, Agrobacterium sp., strain CP4 is not known for human or 

animalpathogenicity, Agrobacterium sp. strain CP4 has been previously reviewed as a 

part of the safety assessment of the donor organism during Monsanto consultations with 

the FDA regarding Roundup Ready soybean (1994), Roundup Ready canola (1995), 

Roundup Ready cotton (1995), Roundup Ready 2 corn (1996), Roundup Ready sugar 

beet (1998), and Roundup Ready Flex cotton (2005). Further, the Environmental 

Protection Agency has established an exemption from the requirement of a tolerance for 

residues of CP4 EPSPS protein and the genetic material necessary for its production in all 

plants (EPA 1996).

(ii) The CP4 EPSPS protein present in MON 88302 is similar to EPSPS proteins 

consumed in a variety of food and feed sources. CP4 EPSPS protein is homologous to 

EPSPS proteins naturally present in plants, including food crops (e.g., soybean and 

maize) and fungal and microbial food sources such as baker’s yeast (Saccharomyces 

cerevisiae), all of which have a history of safe human consumption (Harrison et al. 1996; 

Padgette et al. 1996).

(iii) The CP4 EPSPS protein originates from Agrobacterium sp. strain CP4 an organism 

that has not been reported to be a source of known allergens. Bioinformatics analyses 

demonstrated that the CP4 EPSPS protein does not share amino acid sequence similarities 

with known allergens and, in vitro digestive fate experiments conducted with the CP4 

EPSPS protein demonstrate that the protein is rapidly digested in simulated gastric fluid 

and in simulated intestinal fluid.

(iv) An acute oral toxicology study with mice indicated that the CP4 EPSPS protein did 

not cause any adverse effects in mice at the highest dose level (572 mg/kg) tested 

(Harrison et al., 1996).

(v) The potential route of human exposure to MON 88302 is through canola oil. The 

primary human food currently produced from canola is refined, bleached, and deodorized 

(RBD) oil. Because RBD oil contains negligible amounts of protein (Martín-Hernández 

12
et al., 2008), oil produced from MON 88302 will contain negligible levels of CP4 EPSPS 

protein that would minimize the dietary exposure to this protein from consumption of 

foods derived from MON 88302. Furthermore, the safety of CP4 EPSPS has been 

extensively assessed (Harrison et al., 1996) and several Roundup Ready crops that 

produce CP4 EPSPS have been reviewed by FDA and other regulatory agencies. They 

concluded the Roundup Ready crops were safe for consumption. Likewise Monsanto 

provided data showing that there is minimal exposure to MON 88302 CP4 EPSPS in 

relation to the total protein consumed by animals such as dairy cows, pigs, and chickens 

(FDA BNF No. 127).

Therefore, based on the food and feed safety analyses, including toxicity and allergenicity 

data for CP4 EPSPS, it can be inferred that MON 88302 is unlikely to cause any 

significant adverse effects on nontarget organisms (including beneficial species or 

wildlife associated with canola) compared to other commercial canola varieties. 

Additionally, as indicated earlier in this PPRA, compositional analysis of the seed 

showed that the mean values for nutritional components, anti-nutrients and toxicants 

from the combined site analysis were within the 99% tolerance interval for the 

commercial reference varieties, so there is unlikely to be nontarget effects resulting from 

changes in composition. 

Direct observations detected no differences for beneficial arthropods between MON 

88302 and the conventional control over 15 observations across 4 sites (as summarized in 

Table VII-5, p. 123, Monsanto 2011. Beneficial arthropod observation data were 

collected for Chironomid midge, Lacewings (Chrysopidae), Ladybird beetles 

(Coccinellidae), micro- and macro-parasitic hymenoptera, Orius spp., Spiders (Aranaea), 

and Sphecid wasps (Sphecidae); however with the exception of Orius spp., they were 

rarely observed in either MON 88302, the conventional control, or in the commercial 

reference varieties (Table G10, pp. 335-337). Although honeybees were not assessed in 

the beneficial arthropod study, honeybees tend to show a preference for oilseed rape 

pollen (Cook et al. 2003; Keller et al. 2005). Monsanto did provide observations on 

pollen morphology, and data on pollen diameter and viability from growth chamber 

grown plants showed no significant difference between MON 88302 and the control 

(Appendix H, pp. 339-342, Table H-1, p. 340, Monsanto 2011). Therefore, no direct or 

indirect effects from the transformation process are expected to have an adverse impact 

on beneficial species or wildlife associated with canola compared to other commercial 

canola varieties.

F. Potential for Enhanced Weediness of MON 88302 Canola

APHIS assessed whether MON 88302 canola has attained characteristics as a result of 

genetic engineering that would enhance its weediness compared to the nontransgenic 

progenitor and whether the engineered trait affects methods of control for canola in 

situations where it is managed as a volunteer in subsequent crops or in feral populations. 

13
Canola is a domesticated Brassica species. Canola is not identified as a noxious weed in 

the Federal Noxious Weed List nor does it appear in any state weed lists (USDA-NRCS 

2012). However, canola does possess a few attributes commonly associated with weeds, 

such as a large seed crop and harvest yield loss (Thomas et al. 1991; Brown et al. 1995), 

prolonged seed dormancy of 2-5 years, and an ability to persist as feral populations in 

disturbed habitats (Gulden et al., 2004). Monsanto collected major agronomic data 

relevant to weedy traits such as seed germination, seed dormancy, seedling emergence, 

plant height, lodging, pollen viability and morphology, seed quality and seed moisture, 

seed yield, pod shattering, and abiotic stress tolerance (Table VII-1, p. 109, Monsanto 

2011). Except for seed dormancy, germination, and pollen characteristics investigation, 

all other phenotypic and agronomic data were collected from the experimental set up 

described earlier under the section “Potential Impacts of Genetic Modifications on 

Disease and Pest Susceptibilities.” 

For assessment of the seed germination and dormancy characteristics, the seed lots of 

MON 88302, the conventional control and four commercial reference varieties were 

produced in the field in Grand Forks County, ND in 2009, a geographic area which 

represents an environment with conditions suitable for canola production (Appendix 

Table F-1, p. 309, Monsanto 2011). In addition to the Association of Official Seed 

Analysts recommended temperature regime of 15/25 °C (AOSA 2009a, 2009b), seed was 

tested at five additional temperature regimes of constant 5, 15, 25 or 30 °C, and 

alternating 5/25 °C to assess seed germination properties. The details of the experimental 

materials and methods are presented in Table VII-1 (pp. 108-109, Monsanto 2011) and 

Appendix F (p. 307-311, Monsanto 2011). According to Monsanto, no statistically 

significant differences were detected between MON 88302 and the control at any of the 

temperature regimes for normal and abnormal germination and dead seed and between 

MON 88302 and the control at the 5 oC temperature regime for viable nondormant seed 

(Table VII-2, p. 115, Monsanto 2011). According to Monsanto, statistical comparisons 

were not made due to a low number of dormant seed in the AOSA-recommended 

temperature regime (15/25 oC), MON 88302 numerically had fewer dormant seeds than 

the conventional control (0.0% vs. 0.3%). A reduction in the number of dormant seed 

would not increase plant weediness since non-dormant seed would be more likely to 

germinate reducing the potential for persistence in the soil seed bank (Gulden et al., 

2003). Although, no statistical comparisons were made for viable non-dormant seed in 

the 15 oC, 25 oC, 30 oC and 5/25 oC temperature regimes, the magnitude of the difference 

for seed of MON 88302 and the conventional control was small and there were no 

observable trends in the mean difference in this category across the different temperature 

regimes (p. 114, Monsanto 2011).

The experimental set up and data analysis for agronomic and phenotypic data was 

described earlier in this document (see section “Potential Impacts of Genetic 

Modifications on Disease and Pest Susceptibilities”; the methods and detailed results of 

the individual site data comparisons are presented and discussed in Appendix G, pp. 312-

339, Monsanto 2011). A total of 12 phenotypic and agronomic characteristics were 

evaluated (early stand count, seedling vigor, days after planting to both first flowering 

and seed maturity, lodging, plant height, both visual and quantitative measures for pod 

shattering, seed moisture and quality, yield, and final stand count) (Table VII-4, p. 119, 

14
Monsanto 2011). Only two statistically significant differences were detected between 

MON 88302 and the conventional control in the combined-site analysis: days to 

flowering and seed moisture content (Table VII-4, p. 119, Monsanto 2011). MON 88302 

reached first flowering later than the conventional control (61.1 vs. 56.2 days). However, 

the mean value of MON 88302 for days to first flowering was within the natural 

variability of the commercial reference varieties (45.9 – 67.5 days). Therefore, the 

difference in days to first flowering is unlikely to be biologically meaningful in terms of 

increased weediness potential. Although early flowering is one of the potential indicators 

of enhanced weediness (Campbell et al. 2009; Ridley and Ellstrand 2009), MON 88302 

canola is a late flowering type and the observed difference in flowering is unlikely to 

facilitate any weediness potential to MON 88302 canola. In a separate study on growthchamber grown plants, another reproductive feature, pollen morphology, did not exhibit 

any difference between MON 88302 and control, as no statistically significant differences 

were detected between MON 88302 and the conventional control for percent viable 

pollen or pollen grain diameter (Table VII-7. p. 126, Monsanto 2011). MON 88302 had 

higher harvested seed moisture than the conventional control (13.2% vs. 11.7%). 

However, the mean value of MON 88302 for harvested seed moisture was within the 

natural variability of the commercial reference varieties (7.5% – 14.8%). Therefore, 

according to Monsanto the difference in seed moisture is unlikely to be biologically 

meaningful in terms of increased weediness potential. Likewise, Monsanto did not 

observe significant qualitative differences between MON 88302 and the control canola 

for the nine abiotic stress tolerance traits examined, with the exception of one observation 

for frost damage at one site, which was more severe for MON 88302 but within the range 

observed for the reference varieties (Table VII-5, p. 123, and Table G5, p. 329, Monsanto 

2011). Stress responses examined included cold, heat, frost, hail, drought flood, wind, 

nitrogen deficiency and compaction. 

Seed shattering or seed yield loss during harvest combined with extended seed dormancy 

have the potential to create volunteer and weed problems for subsequent crops. Indeed 

canola is known to shatter seeds with about 2-7% of the seed yield lost during seed 

harvest (Table 2 in Gan et al. 2008). Despite significant seed loss during harvest, a 

majority of fallen seeds in the soil tend to germinate (> 90%) in the first season after 

harvest, and the remaining seeds generally exhibit 1-2 years of dormancy (Gulden et al. 

2003). A few research reports also noted canola seed dormancy periods extending 

beyond 3 years (Légère et al. 2001; Simard et al. 2002; D’Hertefeldt et al. 2008), yet it 

was also observed that very few canola volunteers emerge during and after the third year 

of the post-harvest (Simard et al. 2002). Nonetheless, volunteer canola plants have still 

been documented at low densities four and five years after production (Simard et al. 

2002). A significant body of research exists on the ability of canola to form feral 

populations (Simard et al. 2002 and references therein; Schafer et al. 2011) and there is a 

widespread concern that herbicide-resistant feral populations may become an 

unmanageable weed problem around field edges and minimally managed 

agroecosystems. Unlike highly domesticated crops such as corn and soybean, canola is a 

relatively newly domesticated crop plant and possesses a few traits (e.g. prolonged seed 

dormancy, large seed yield, seed shattering) that facilitate canola to persist as feral 

populations (Crawley and Brown 1995; Pessel et al. 2001). On the contrary, a mere 

15
possession of the potential weedy traits (Baker 1965) does not appear to predispose a 

plant taxon to become a weed (Perrins et al. 1992; Sutherland 2004). 

Despite possessing some of the weed traits, canola is unlikely to become an 

unmanageable weed with the introduced trait. Like other crop plants, canola has several 

domesticated traits such as high seed output under optimum agronomic practices, selfpollination, etc., that make canola less competitive in unmanaged or minimally managed 

ecosystems (Crawley et al. 1993; Crawley et al. 2001; Salisbury 2002). The agronomic 

characteristics and germination data discussed earlier in this section provide evidence that 

the genetic modification resulting in MON 88302 canola did not alter any major 

characteristics of the plant that would allow for development of weedy characteristics 

different from other canola varieties. Furthermore, the herbicide-resistance trait conferred 

by the cp4 epsps gene is unlikely to provide a selective advantage in unmanaged 

ecosystems, but rather only in settings where glyphosate is being applied for weed 

control. 

As described in the petition (section VII.D., p. 126, Monsanto 2011), herbicide-resistant 

canola is no more likely to form feral populations than unmodified canola, nor is it more 

likely to be more invasive or competitive or persistent in habitats where the target herbicide 

is not applied (Warwick et al. 2009; Andersson and de Vicente 2010). Even in those areas 

where herbicide-resistant canola has been grown extensively in the last several years, there is

no indication of altered weediness or invasiveness potential imparted to feral canola or 

volunteer canola populations (Hall et al. 2005). 

Volunteer herbicide-resistant canola (MON 88302 canola, as well as other commercially 

available glyphosate, glufosinate, and imidazolinone resistant varieties) should be controlled 

before planting canola cultivars with different herbicide resistance traits to reduce the 

potential for gene flow to result in stacked herbicide resistance, as such stacked herbicide 

resistance in canola has already been documented (Hall et al. 2000; Shaefer et al. 2011) 

(discussed in more detail in the following section). Although herbicide-resistant and 

nonresistant canola varieties are documented to persist as volunteers or establish feral 

populations, there are alternative herbicides that can be used to control them. Paraquat 

and diuron (alone or in various combinations, some including glyphosate) were shown to 

be effective herbicides for control prior to planting in conservation tillage systems in the 

Pacific Northwest (Rainbolt et al. 2004). Producers need to consider which crop will 

follow the canola when making herbicide selections. Canola best follows cereal grains or 

fallow in rotation, and is rarely planted within one or two years following canola and 

other crops highly susceptible to sclerotinia (a stem rot) e.g. sunflower, dry edible beans 

or crambe, and sugar beet (Berglund et al. 2007). Glyphosate resistant canola is typically 

rotated with other crops, typically wheat in a two year rotation or with wheat and soybean 

in a three year rotation, and other rotation crops include oats, barley, and flax (Berglund 

et al. 2007). Several (26) herbicide formulation options provide at least good to excellent 

control of volunteer glyphosate resistant canola, particularly at the 3-6 leaf stage, as 

described in the 2012 North Dakota Weed Control Guide, and several can be used in the 

most common rotation crops of canola (NDSU 2012, p. 115; Table VIII-12, p. 164, 

Monsanto 2011). Tillage can also be used. CP4 EPSPS does not confer cross resistance 

16
to other herbicide modes of action. Therefore, MON 88302 is expected to be sensitive to 

the same herbicides as other glyphosate resistant canola already commercialized. 

Therefore, based on this characterization, MON 88302 is no more likely to establish feral 

populations than either existing transgenic or nontransgenic herbicide-resistant or 

nontransgenic herbicide sensitive canola varieties, and such feral populations can be 

controlled using current weed control practices.

G. Potential of MON 88302 to Impact the Weediness of Other Plants 

with which It Can Interbreed

Gene flow is a natural biological process with significant evolutionary importance. A 

number of angiosperm taxa are believed to be derived from hybridization or introgression 

between closely related taxa (Grant 1981; Soltis and Soltis 1993; Rieseberg 1997; Hegde 

et al. 2006), and even in the existing floras, the occurrence of hybridization or 

introgression is reported to be widespread (Stace 1987; Rieseberg and Wendel 1993;

Peterson et al. 2002). It has been a common practice by plant breeders to artificially 

introgress traits from wild relatives into crop plants to develop new cultivars. However, 

gene flow from crops to wild relatives is also thought of as having a potential to enhance 

the weediness of wild relatives, as observed in rice, sorghum, sunflower and a few other 

crops (see Table 1 in Ellstrand et al. 1999). 

APHIS considers two primary issues when assessing weediness of sexually compatible 

plants because of transgene flow: 1) the potential for gene flow and introgression and, 2) 

the potential impact of introgression. 

Canola is predominantly self-pollinating, but outcrossing does occur via wind and insect 

pollination (William 1984; William et al. 1987). Depending on the size of the crop and 

distance between plants or fields, a variety of outcrossing rates (12-55%) were observed 

for canola (See Table 1 in Beckie et al. 2003). Most outcrossing between fields generally 

occurs within the first 10-20 m of the recipient field, and rates decline with distance 

(Table 1 in Beckie et al. 2003; Table IX-2, p. 181, Monsanto 2011). 

Canola is grown in very few places in the U.S. and a majority (~88%) of canola 

production occurs in North Dakota, while the remaining cultivation comes from 

Minnesota, Idaho, Washington, Montana, Oklahoma, and Oregon (USDA-NASS 2011; 

Table VIII-2, p. 132, Monsanto 2011). Spring and winter canola varieties are generally 

grown in different regions of the U.S. based on climate zones most suitable for the 

varieties (pp. 129-130 and 138 in Monsanto 2011). Brassica crops involve a number of 

diploid and polyploidy species and the family Brassicaceae involves a number of major 

weed species (OECD 2012). 

In a majority of crop species, gene flow is idiosyncratic depending upon biology and 

ecology of both crop and sexually compatible relatives (Gliddon et al. 1999; Ingram 

2000; Warwick et al. 2009). Accordingly, there are several important considerations for 

a successful gene flow and introgression between MON 88302 and sexually compatible 

crop and weedy relatives such as spatial proximity, overlapping phenology, F1 hybrid 

17
fertility, self-sustaining reproductively fertile hybrid-derived (backcrossed) populations, 

and neutral or beneficial introgressed genes (Devos et al. 2009). In addition, the stage at 

which the crop is harvested, e.g. before or after flowering or seed set, will also influence 

the likelihood of successful gene flow.

Monsanto provided a summary of published literature on unassisted hybridization under 

field conditions with B. napus as a the male parent to various crops and wild relatives of 

canola that occur in the U.S. (Table IX-3, p. 189-193, Monsanto 2011) and the likelihood 

of hybridizations with canola based on information on the success rate of hand 

pollinations or spontaneous and natural hybridization, weediness, and presence of the 

species in winter or spring canola growing areas (section IX.D., pp. 180-193, Monsanto

2011). From this analysis Monsanto concludes: “There are reports of hybridization under 

field conditions with B. napus as the pollen donor with six species including Brassica

rapa, B. juncea, B. oleracea, Hirschfeldia incana, Raphanus raphanistrum and Sinapsis

arvensis (Table IX-3). The species B. rapa, B. juncea and B. oleracea are cultivated for 

crop production. The other species listed, H. incana, R. raphanistrum and S. arvensis are 

not cultivated for crop production, but are found in the environment. In all cases the 

resulting hybrids had decreased environmental fitness evidenced by a variety of 

characteristics including decreased pollen viability, seed production, seedling survival, 

etc. when compared to parental varieties.” (p. 182, Monsanto 2011). APHIS agrees with 

this conclusion, which is well supported by published literature.

Genus Brassica and related genus Raphanus contain oil seed, vegetable, and forage crop

species (Ellstrand 2003; FitzJohn et al. 2007) such as B. napus (oil seed, swede) and B. 

rapa (oilseed, turnip and Chinese cabbage), B. oleraceae (cauliflower, cabbage, 

broccoli), B. juncea (Indian mustard), and R. sativus (radish). Three Brassica species and 

one species in the related genus Sinapis are ‘mustards’: B. carinata (Ethiopian mustard), 

B. juncea (Indian mustard), B. nigra (black mustard) and Sinapis alba (white mustard). 

Cultivation of B. carinata as an oilseed and vegetable crop is largely restricted to 

Ethiopia and India (Hemingway 1995; Stewart 2002). Some forms of B. napus, B. 

oleracea, B. rapa and R. sativus are also grown as fodder crops (FitzJohn et al. 2007).

The three Brassica species forming the foundation of the Triangle of U showing genome 

relationships among cultivated Brassicaceae (Nagaharu 1935) are B. rapa, B. nigra, and 

B. oleracea. B. napus and B. rapa outcross readily with each other, while Brassica napus 

and B. juncea share a common set of chromosomes, enhancing the likelihood of 

interspecific hybridization and gene flow (Myers 2006). The A genome is common to 

the three major oilseed Brassica species, which explains the success of interspecific 

crossing, and the ability to transfer genes among these species. 

The Brassicaceae family contains a number of major weeds, including those in the genera 

Sinapis, Capsella, Thlaspi, Erucastrium, Raphanus, and others (OECD 1997). Concerns 

have been raised about the potential for the transfer of transgenes from the cultivated 

oilseed Brassica species to their weedy relatives in Europe and North America where 

Brassica crop species are widely grown. These Brassica crop species can also outcross, 

albeit rarely, with a wide range of wild and weedy species (summarized in OGTR 2002). 

Some Brassica crops and their wild relatives will hybridize only under artificial 

18
conditions in laboratories or highly contrived field conditions; whereas others will 

hybridize at very low rates under natural conditions (Raybould 1999; Barton and Dracup 

2000). Through an extensive literature survey, Warwick and Black compiled an 

exhaustive list of interspecific and intergeneric hybridization among the members of the 

tribe Brassiceae, including large-scale artificial intergeneric hybridizations between 

various members of the tribe (Table 1 in Warwick and Black 1993) and reported very few 

natural hybrids. As noted earlier, several reproductive and ecological barriers between 

canola and its wild species prevent formation of successful introgressed, self-sustaining 

hybrid derived populations (see detail descriptions in section IX.D., pp. 180-193 in 

Monsanto 2011). 

Feral canola is a quite common occurrence along canola field edges and transportation 

routes (Bagavathiannan and Van Acker 2008; see Table 1 in Devos et al. 2012). The major 

concern raised about herbicide-resistant GE canola is about the ability of feral canola 

populations to act as a sink for the accumulation (stacking) of multiple herbicide 

resistance genes from the four different herbicide-resistant (GE-glufosinate resistance, 

GE-bromoxynil resistance, GE-glyphosate resistance, non-GE imidazolinone-resistant) 

canola currently under cultivation in Canada (Hall et al. 2000). There are reports 

indicating that canola cultivars with different HR traits resulted in volunteers with 

multiple resistance at a field site in western Canada (Hall et al. 2000). Therefore, the 

stacking of HR traits has the potential to facilitate the evolution of invasive feral canola 

populations (Ellstrand 2003; Knispel et al. 2008; Warwick et al. 2009). 

In 1997 in northern Alberta, a field of glyphosate resistant canola was grown adjacent to 

a field of glufosinate resistant and imidazolinone resistant canola. Volunteers were 

selected with glyphosate in 1998 (Hall et al. 2000). These volunteers flowered and 

produced seeds that contained individuals resistant to glyphosate and glufosinate; 

glyphosate and imazethapyr; and glyphosate, imazethapyr, and glufosinate. Two triple 

herbicide-resistant individuals were detected, with one plant located 550 m from the 

glyphosate resistant pollen source. More recently, a study at 11 sites in Saskatchewan, 

Canada, where glyphosate HR B. napus canola was grown adjacent to glufosinate HR B. 

napus canola, documented gene flow to the limits of the study areas—a maximum 

distance of 800 m—on the basis of occurrence of double-HR volunteers (Beckie et al. 

2003). The results of both studies suggest that herbicide-resistant gene stacking is 

common in B. napus canola volunteers in western Canada. In a similar study Knispel et al 

(2008) surveyed for the presence of single and multiple herbicide resistance traits and 

assessed the extent of gene flow within escaped canola populations. Seed was collected 

from 16 escaped canola populations along the verges of fields and roadways in four 

agricultural regions in southern Manitoba from 2004 to 2006. Glyphosate resistance was 

found in 14 (88%) of these populations, glufosinate resistance in 13 (81%) populations, 

and imidazolinone resistance in five (31%) populations. Multiple herbicide resistance 

was observed at levels consistent with previously published canola outcrossing rates in 10 

(62%) of the tested populations. These reports indicate that intraspecific gene flow 

results in stacking of herbicide resistance traits in individuals within escaped canola 

populations (Knipsel et al. 2008). Similar multiple herbicide-resistant canola feral 

populations were also reported from Japan around transportation routes, although Japan 

19
never cultivates GE canola varieties and only imports them for food and feed purposes 

(Aono et al. 2006; Kawata et al. 2009).

Gene flow from MON 88302 was evaluated thoroughly with respect to plant pest risk. 

The introduced cp4 epsps gene in MON 88302 is not expected to change the ability of the 

plant to interbreed with other plant species. Furthermore, the APHIS-BRS evaluation of 

data provided by Monsanto (2011) of agronomic and phenotypic properties of MON 

88302, including those characteristics associated with reproductive biology, indicated no 

unintended changes likely to affect the potential for gene flow from MON 88302 to 

sexually compatible species. In addition, gene flow has been occurring between non-GE 

canola (both herbicide-resistant and other canola varieties) and sexually compatible 

species even before the advent of GE canola varieties. Therefore, the consequences of 

gene flow and introgression of the glyphosate-resistant trait from MON 88302 to the 

same or sexually compatible species are anticipated to be the same as for existing 

commercial glyphosate-resistant canola varieties.

Successful hybridization of canola and a wild/weedy relative is highly unlikely and even 

if those successful rare events occur, the herbicide-resistance trait would only provide 

selective advantage in situations in which the weedy hybrid was in contact with the 

herbicide (i.e., in an agricultural field or treated rights of way). One such situation is 

when feral canola populations or wild relatives of canola are exposed to herbicide due to 

herbicide drift, whereby some herbicides are carried by winds and airflow beyond the 

intended area of application. Londo et al. (2010) demonstrated the potential for changes 

in the movement and persistence of glyphosate- resistance transgenes in weedy plant 

communities due to gene flow and fitness traits under the presence of glyphosate drift. 

As authors noted in their research paper, the above-mentioned research results were 

obtained from outdoor sunlit mesocosms (simulated and controlled natural system) and 

were not performed as a field experiment. Furthermore, Londo et al. (2010) advocated 

for future field-based studies to validate their observations and to test for other 

environmental factors that influence the gene flow dynamics of weedy populations 

exposed to glyphosate drift. Therefore, so far there is no strong evidence that genetically 

engineered glyphosate herbicide-resistant canola has substantially altered the gene flow 

dynamics between GE canola and its wild and weedy relatives.

Any herbicide-resistant feral and hybrid-derived populations are likely to be controlled 

using other available chemical or mechanical means. Many herbicides that are effective 

for control of glyphosate resistant canola are also effective for control of wild mustards 

(NDSU 2012, pp. 115, 117, 119, and 120). As described by Beckie and colleagues (see 

Beckie et al. 2004 and references therein) the following cultural or mechanical practices 

are recommended to growers to manage multiple-HR canola volunteers: (1) leaving seeds 

on or near the soil surface as long as possible after harvest because a high percentage will 

germinate in the fall and be killed by frost, whereas seeds incorporated into the soil may 

develop secondary dormancy that will increase persistence; (2) using tillage immediately 

before seeding; (3) silaging and green manuring to prevent seed set in volunteers; (4) 

isolating fields of canola with different herbicide resistance traits to reduce outcrossing; 

(5) following canola with a cereal crop and rotating canola in a 4-yr diverse cropping 

sequence to deplete volunteers from the seedbank over time (which also facilitates use of 

20
alternative herbicides with different modes of action) and growing competitive crops to 

minimize volunteer canola interference (by choice of species and manipulation of 

agronomic practices such as higher seeding rates and precision fertilizer placement); (6) 

scouting fields for volunteers not controlled by weed management treatments and 

preventing seed set; (7) using pedigreed seed to reduce the probability of the presence of 

off-types with different herbicide resistance traits; and (8) reducing seed loss during 

harvest by swathing at the correct crop development stage and properly adjusting 

combine settings. Herbicide treatments such as metribuzin, 2,4-D, or MCPA, alone or in 

a mixture, can control single or multiple herbicide-resistant canola volunteers when 

densities warrant, either pre-seeding or in-crop where registered. Previous studies have 

shown no difference in fitness among non-herbicide, single herbicide-resistant, or 

multiple herbicide-resistant canola. These findings indicate that multiple herbicideresistant canola can be controlled equally well as non-herbicide-resistant or single 

herbicide-resistant plants by alternative herbicides within an integrated weed 

management program (Beckie et al. 2004).

Large-scale cultivation of herbicide-resistant canola has occurred for nearly 15 years in 

Canada and the United States. To date, there are no reports of problems with interspecific 

crosses and introgression of herbicide-resistance genes into cultivated or wild relatives of 

canola (Andersson and de Vicente 2010). The International Survey of Herbicide 

Resistant Weeds has no confirmed cases of glyphosate resistant weeds that are wild or 

weedy relatives of canola (Heap 2013). The MON 88302 is not expected to expand the 

amount of acreage planted to canola or to glyphosate resistant canola, but rather to 

provide growers an opportunity to control troublesome weeds of canola crops with the 

glyphosate herbicide application during later stages of plant development than currently 

possible with other varieties of glyphosate-resistant canola already commercially 

available. 

Therefore, it is highly unlikely that canola plants in the United States will be found 

outside of an agricultural setting, except along roadsides along seed transportation routes. 

It is also highly unlikely that gene flow and introgression will occur between MON 

88302 plants and wild or weedy species in a natural environment. Herbicides are 

available to control volunteer glyphosate-resistant canola and weedy relatives. USDA 

has therefore determined that any adverse consequences of gene flow from MON 88302

to wild or weedy species in the United States are highly unlikely.

H. Potential Changes to Agriculture or Cultivation Practices

APHIS considered whether there are likely to be significant changes to agricultural 

practices associated with cultivation of MON 88302, and if so are they likely to 

significantly exacerbate plant diseases or pests, especially those for which APHIS has a 

control program. 

Glyphosate-resistant MON 88302 is not a new type of GE crop, as three different types of 

herbicide-resistant canola varieties (conventionally derived imidazolinone-resistant 

(Clearfield); GE glyphosate-resistant (Roundup Ready RT73); and GE glufosinateresistant (InVigor)) are already available for cultivation in the U.S. (Brown et al. 2008). 

21
In addition, a significant acreage of genetically modified herbicide-resistant canola 

(predominantly glyphosate-resistant) has been planted commercially in the U.S. since 

1999. For instance, in 2008, genetically engineered herbicide-resistant canola was 

estimated to be 95% of the U.S. canola crop (Brookes and Barfoot 2010). One major 

difference between MON 88302 and earlier glyphosate resistant canola varieties is that 

MON 88302 is modified to withstand a later application of glyphosate compared to the 

earlier herbicide-resistant canola varieties. According to Monsanto, MON 88302 is a 

second-generation glyphosate-resistant spring canola variety designed to provide growers 

with improved weed control through greater flexibility of glyphosate herbicide 

application. By virtue of enhanced CP4 EPSPS expression in male reproductive tissues, 

MON 88302 provides tolerance to glyphosate during the sensitive reproductive stages of 

growth, and enables the application of glyphosate at later stages of development and at 

higher rates than is possible with the current product (Feng et al. 2010; p. 196 in 

Monsanto 2011). 

Current label directions indicate glyphosate agricultural herbicides can be applied postemergence on glyphosate-tolerant canola from emergence to the 6-leaf stage of 

development up to 0.39 to 0.56 lb a.e./acre in spring canola varieties and 0.77 lb a.e./acre 

for winter canola varieties to prevent weed competition. The first application of 

glyphosate to glyphosate-tolerant canola is made at the two- to three-leaf stage to prevent 

early weed competition and an additional application is made up to the six-leaf stage to 

control any late-emerging weeds. Two applications of glyphosate were made on 

approximately 42% of the canola acreage in the largest canola producing state, North 

Dakota, in 2008 (Table VIII-8, p. 153, Monsanto 2011), however, after the 4-leaf stage a 

single application of glyphosate greater than 0.39 lb a.e./acre in spring canola and greater 

than 0.56 lbs a.e./acre prior to the 6-leaf stage in the fall for winter canola can result in 

crop injury. In winter canola, a sequential application up to 0.77 lbs a.e./acre can be made 

the following spring prior to the bolting stage, as long as the two in-crop applications do 

not exceed a total of 1.55 lbs. a.e./acre (Section VIII.G., pp. 156-157, Monsanto 2011).

MON 88302 provides greater crop tolerance which will permit higher rates and a wider

period for application of glyphosate in spring and winter canola compared to the firstgeneration Roundup Ready canola system. The higher glyphosate rates and extended 

timing for applications up to the first flower stage will provide improved control of 

difficult to control weed species such as Canada thistle, dandelion, sow thistle, common 

lambsquarters, kochia, smartweed and wild buckwheat (CCC 2003), however glyphosate 

resistant kochia has now been reported in crop land in six states (including significant 

canola-producing states, North Dakota and Montana, as well as in Alberta, Canada (Heap 

2013). If the glyphosate label is amended by the EPA, MON 88302 will permit two incrop sequential glyphosate applications up to 0.77 lbs. a.e./ acre each in spring canola or 

one application up to 1.55 lb a.e./acre prior to the 6-leaf growth stage. Total in-crop 

applications will be increased from 0.78 to 1.55 lbs of glyphosate a.e./acre for spring 

canola. Recommended application rates for glyphosate in winter canola will likely 

remain unchanged with the introduction of MON 88302 (Section VIII.G., p. 157, 

Monsanto 2011). Although MON 88302 commercial cultivation would likely increase 

the total amount of the glyphosate herbicide use in canola crops for an extended period of 

time during crop cultivation, similar cultivation practice has already been in use with 

22
several GE herbicide-resistant crops that are planted every year across millions of acres

of agricultural lands. Moreover, the increased glyphosate use on MON 88302 would be 

negligible (1%) compared to the amount of glyphosate usage on other glyphosate 

herbicide-resistant crops such as corn, cotton, and soybean which are grown over much 

larger acreages (NDSU 2007; Benbrook 2009). Issues related to weed control and 

herbicide resistance as a result of changes in herbicide use are covered in the NEPA 

document prepared for this petition.

Recent studies (Anderson and Kolmer 2005; Feng et al. 2005, 2008) have reported that 

glyphosate is active against certain fungal diseases (e.g. in wheat and soybean) and when 

applied to GR crops provided disease suppression that benefit from the persistence of 

glyphosate. As this is primarily a glyphosate effect, this issue will be more fully 

evaluated in the NEPA document prepared for this petition. A more recent review by 

Duke et al. (2012) indicates that although it is clear that glyphosate does increase severity 

of disease on glyphosate susceptible plants, overall the baseline disease resistance or 

susceptibility of the host plant, not the presence of the glyphosate resistance gene or 

treatment with glyphosate, is the major contributor to susceptibility. None of the 

referenced studies on disease response following glyphosate treatment in glyphosate 

resistant crops included glyphosate resistant canola.

As discussed throughout this document, MON 88302 is similar to conventional canola in 

its agronomic, phenotypic, environmental intercalations, and compositional 

characteristics and has levels of tolerance to insects and diseases comparable to 

conventional canola. Therefore, no significant impacts on current cultivation and 

management practices for canola are expected following the introduction of MON 88302.

I. Potential for Transfer of Genetic Information to Organism with 

which MON 88302 Canola Cannot Interbreed and Potential Impacts 

of Such Transfer

The horizontal gene transfer (HGT) between unrelated organisms is one of the most 

intensively studied fields since 1940, and the issue gained extra attention with the release 

of transgenic plants into the environment (Dröge et al. 1998) and sequencing of large 

numbers of genomic sequences (Choi and Kim 2007). HGT contributed to major 

transitions in evolution of prokaryotic organisms (Woese 2002) and has been implicated 

as a major contributor to the spread of antibiotic resistance amongst pathogenic bacteria 

and the emergence of increased virulence in viruses, bacteria, and eukaryotes. Although, 

gene exchange has been documented for nearly all types of genes and between unrelated 

organisms at an evolutionary scale (Gogarten et al. 2002; Yoshida et al. 2010), the 

frequency of HGT among higher organisms are shown to be extremely rare and, 

consequently, such transfers did not play any major role in their evolution (Kurland et al. 

2003).

APHIS examined the potential for the new genetic material inserted into MON 88302 to 

be horizontally transferred to other organisms without sexual reproduction and whether 

such an event could lead directly or indirectly to disease, damage, injury or harm to 

plants. Although MON 88302 contains genetic sequences from the plant pathogenic 

23
organisms Agrobacterium and figwort mosaic virus, the expressed gene product does not 

have plant pathogenic properties or cause disease in plants. Furthermore, many 

organisms, including pathogens and symbionts, already contain variations of genes 

encoding EPSPS with varying degrees of sensitivity to glyphosate (see Duke et al. 2012). 

Furthermore, horizontal gene transfer and expression of DNA from a plant species to 

other fungal, bacterial, or parasitic species is unlikely to occur based on the following 

observations.

Although there are many opportunities for plants to directly interact with fungi, bacteria, 

and parasitic plants (e.g. as commensals, symbionts, parasites, pathogens, decomposers, 

or in the guts of herbivores), so far there are no reports of significant horizontal gene 

transfer between evolutionarily distant organisms (as reviewed in Kurland et al. 2003; 

Keese 2008). Accumulated evidence show that there are universal gene-transfer barriers, 

regardless of whether transfer occurs among closely or distantly related organisms 

(Koonin et al. 2001; Wood et al. 2001; Kaneko et al. 2002; Brown 2003; Sorek et al. 

2007). Many genomes (or parts thereof) have been sequenced from bacteria that are 

closely associated with plants including Agrobacterium and Rhizobium (Kaneko et al. 

2002; Wood et al. 2001). There is no evidence that these organisms contain genes 

derived from plants. In cases where review of sequence data implied that horizontal gene 

transfer occurred, these events are inferred to occur on an evolutionary time scale on the 

order of millions of years (Koonin et al. 2001; Brown 2003), so also the case with the 

recent report about HGT between sorghum and purple witchweed. According to authors 

(Yoshida et al. 2010), the incorporation of a specific genetic sequence occurred between 

sorghum and purple witchweed before speciation of purple witchweed (Striga

hermonthica) and related cowpea witchweed (S. gesnerioides), a parasitic plant of dicots, 

from their common ancestor. In other words, HGT is an extremely rare event, and a 

majority of those rare events occur over millions of years. 

Transgene DNA promoters and coding sequences are optimized for plant expression, not 

prokaryotic bacterial expression. Thus even if horizontal gene transfer occurred, proteins 

corresponding to the transgenes are not likely to be produced. FDA has evaluated 

horizontal gene transfer from the use of antibiotic resistance marker genes and concluded 

that the likelihood of transfer of antibiotic resistance genes from plant genomes to 

microorganisms in the gastrointestinal tract of humans or animals, or in the environment, 

is remote (FDA 1998: http://www.fda.gov/Food/GuidanceComplianceRegulatory

Information/GuidanceDocuments/Biotechnology/ucm096135.htm). Therefore APHIS 

concludes that horizontal gene transfer is unlikely to occur from MON 88302 to 

microorganisms, and thus no significant plant pest risk is expected from horizontal gene 

transfer.

J. Conclusion

APHIS has reviewed the information submitted by the petitioner and conducted a plant 

pest risk assessment on MON 88302 (including its progeny). APHIS concludes that 

MON 88302 canola is highly unlikely to pose a plant pest risk for the following reasons:

 

24
a. Neither the introduced sequences nor the method of transformation has resulted in 

disease symptoms, pathogen infection, or expression of a pathogen in MON 

88302. 

b. Composition of Mon 88302 seed is similar to that of the conventional control 

and/or within the natural variability of commercial reference varieties. Therefore, 

changes in gene expression, enzymes or metabolism from introduced genes in 

MON 88302 are unlikely to pose a plant pest risk.

c. MON 88302 is expected to be susceptible to the same plant pathogens and insect 

pests as conventional canola and it is unlikely to bring about any indirect plant 

pest effects on other agricultural products. 

d. MON 88302 is not expected to adversely impact wildlife or other organisms 

beneficial to agriculture any more than conventional canola varieties. 

e. Introduced genes in MON 88302 did not significantly alter any major 

characteristics of the plant (i) that would facilitate the development of weedy 

characteristics in the crop or (ii) that would enhance its gene flow potential to 

wild/weedy relatives and consequently produce or alter weedy characteristics in 

wild/weedy relatives. 

f. The glyphosate resistance trait in MON 88302 and the anticipated changes in 

agricultural practices related to glyphosate resistance in MON 88302 are not 

expected to increase pests or diseases or impact their control in canola or other 

crops based on prior experience with previously deregulated glyphosate resistant 

canola. 

g. Genes encoding variants of the EPSPS protein already exist among pathogens and 

symbionts in the environment, and horizontal transfer of the inserted glyphosate 

resistance gene from MON 88302 to other organisms with which it cannot 

interbreed is highly unlikely, and thus should not pose a plant pest risk.

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