Plant Pest Risk Assessment for Syngenta COT67B Cotton

Syngenta Biotechnology, Inc., (referred to hereafter as Syngenta) has petitioned the 

Animal and Plant Health Inspection Service (APHIS) of the United States Department of 

Agriculture for a determination that genetically engineered (GE) cotton (Gossypium 

hirsutum) event COT67B (APHIS number 07-108-01p) is unlikely to pose a plant pest

risk and, therefore, should no longer be a regulated article under APHIS’ 7 Code of 

Federal Regulations (CFR) part 340. APHIS administers 7 CFR part 340 under the 

authority of the plant pest provisions of the Plant Protection Act of 20001. This plant pest 

risk assessment was conducted to determine if COT67B cotton is unlikely to pose a plant 

pest risk. 

Potential impacts to be addressed in this risk assessment are those that pertain to the use 

of COT67B and its progeny in the absence of confinement. APHIS utilizes data and 

information submitted by the applicant, in addition to current literature, to determine if 

COT67B is unlikely to pose a plant pest risk. If APHIS determines that a GE organism is 

unlikely to pose a plant pest risk, then APHIS has no regulatory authority over that 

organism. 

Based on the information requested by APHIS for submission of a petition for 

determination of nonregulated status (§ 340.6(c)) and in order to determine whether 

COT67B is unlikely to pose a plant pest risk, BRS considered information such as: plant 

pest risk characteristics, disease and pest susceptibilities, expression of the gene product, 

new enzymes, changes to plant metabolism, weediness of the regulated article, impact on 

the weediness of any other plant with which it can interbreed, agricultural or cultivation 

practices, effects of the regulated article on non-target organisms, and transfer of genetic 

information to organisms with which it cannot interbreed.

The Environmental Assessment (EA) for this petition included an analysis on agricultural 

or cultivation practices for COT67B. Potential impacts addressed in this risk assessment 

are those that pertain to plant pest risk characteristics. The GE construct inserted in

COT67B was evaluated to determine if the sequences inserted into the cotton could cause

plant disease. In addition, morphological characteristics of this cotton were analyzed to 

determine if this variety would become weedy or invasive. Gene flow and introgression 

of the inserted genes into weedy and wild relatives was evaluated to determine the 

potential of increased weedy or invasive characteristics; also, the potential to transfer 

genetic information to organisms with which cotton cannot interbreed. Finally, APHIS 

evaluated and compared COT67B to conventional cotton in regards to disease and pests 

susceptibility and conducted an analysis of the effects of the regulated article on nontarget organisms. A thorough assessment of the effects of the determination on non-

 1

Section 403 (14) of the Plant Protection Act (7USC Sec 7702(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.” 
Page 2 of 16

target and beneficial organisms, and threatened and endangered species was included in 

the EA.

Development of Lepidopteran Resistant Cotton event COT67B

Cotton growers in the United States, the world’s largest exporter of cotton, suffer

significant yield loss due to insect pests (NCC 2008). There is a need to provide growers 

with a cost effective method to control insects, while taking into account the potential 

hazardous nature of insecticides. GE cotton expressing Cry proteins derived from 

Bacillus thuringiensis (Bt), commonly referred to as Bt cotton, have a long history of safe 

use without adverse human health or environmental effects and provide an option for the 

control of lepidopteran insect pests (Glare 2000, Romeis 2008, NAS 2004).

Syngenta has genetically engineered G. hirsutum (referred to as Upland cotton or cotton)

to express a Cry1Ab protein for use in the control of lepidopteran pests. COT67B deters 

feeding by several insects known to cause significant damage to cotton in the U.S. These 

include: tobacco budworm (Heliothis virescens), cotton bollworm (Helicoverpa zea), 

pink bollworm (Pectinophora gossypiella), and cabbage looper (Trichoplusia ni) (USDAAPHIS 2008, Williams 2008).

GE cotton containing insect-resistant traits, first approved for commercial production in 

the U.S. in 1996, has been widely adopted by U.S. growers (USDA-AMS 2009, USDAERS 2009). Most of the growers who adopt Bt cotton do so mainly to increase yields 

through improved pest control (USDA-ERS 2006). Plantings of GE insect resistant 

cotton increased rapidly in the U.S. to the current 65 percent (Figure 1) (USDA-ERS 

2009). Transgenic varieties - GE varieties resistant to worms, herbicides, or both -

accounted for 88 percent of the Upland cotton planted in the U.S. in 2009 (USDA-ERS 

2009). Grower benefits are likely to be higher with Bt cotton in areas and years with high 

infestation levels of the target pests (EPA 2008, USDA-ERS 2006).
Page 3 of 16

Figure 1. Adoption rates of genetically engineered crops in the U.S. (USDA-ERS 2009).

Tolerance exemptions and conditional pesticide registrations have been granted for the 

plant-incorporated protectant in COT67B and the genetic material necessary for its 

production. On July 16, 2008, the Environmental Protection Agency (EPA) granted an 

exemption from the requirement of a tolerance for residues of Cry1Ab in or on food and 

feed commodities of cotton (73 FR 40760-40764). Likewise, on October 29, 2008, EPA 

approved the conditional registration of Cry1Ab produced in COT67B for use as a 

lepidopteran insecticide (73 FR 64323-64324). Syngenta’s food safety summary 

submitted to FDA indicated that food and feed derived from COT67B are as safe and 

nutritious as those derived from conventional cotton. Following this voluntary 

biotechnology consultation, on February 13, 2009, the FDA concluded that it had “no 

further questions concerning food and feed derived from cotton event COT67B” (FDA 

BNF No. 0112).

Description of the Modification

COT67B has been genetically engineered to contain one transgene construct2: an actin

promoter, a full-length cry1Ab gene, and a nopaline synthase terminator, and non-coding 

regulatory regions.

 2 The aph4 gene from Escherichi coli, encoding for the selectable marker hygromycin-B 

phosphotransferase, was also incorporated as part of the initial transformation process. However, the 

transformation process resulted in the introduction of the cry1Ab gene and the aph4 gene into two different 

chromosomes. The presence of the aph4 gene allowed for selection of transformed cells containing cry1Ab

gene in the presence of hygromycin. As part of the breeding process to develop COT67B, the aph4 gene 

was segregated out, and is not present COT67B (Syngenta 2007, Figures 3-13 through 3-16). 
Page 4 of 16

• The actin promoter and intron from the actin-2 gene of Arabidopsis thaliana (An 

1996). 

• The full-length cry1Ab gene from B. thuringiensis subsp. kurstaki strain HD-1

encodes the Cry1Ab protein. COT67B contains a full-length cry1Ab gene 

(flcry1Ab, 3546bp) which encodes a full-length crystal protein comprising 1181 

amino acids (Syngenta 2007, Appendix 1-Figure1). Full-length cry1Ab in 

COT67B was originally derived from Bacillus thuringiensis subsp. kurstaki strain 

HD-1 (Btk). The native or wild-type cry1Ab gene is a product of the genetic 

recombination of the B. thuringiensis kurstaki HD-1 genes, cry1Aa and cry1Ac. It 

is well documented that mutations resulting in deletions of stretches of DNA are 

common during genetic recombination. Such was the case for the native cry1Ab 

gene (Geiser 1986). This loss of DNA resulted in a diminished capacity of native 

cry1Ab to encode Cry1Ab protein in fermentative cultures of B. thuringiensis

under the customary fermentation temperatures. It was subsequently discovered 

that this inefficiency was attributable to the absence of 26 amino acids encoded by 

the stretch of DNA lost as a result of recombination. Syngenta “repaired” the 

gene by replacing the deleted coding region with the functional region from 

cry1Aa, thus restoring the original fermentative properties of cry1Ab. The 

additional 26 amino acids in the C-terminal portion of the FLCry1Ab protein are 

referred to as the ‘Geiser motif’ (Geiser 1986, 1991; Koziel 1997) and are 

encoded by the “full-length” cry1Ab (flcry1Ab) gene. This “full-length” version 

of the gene was used in transformation to produce COT67B and is referred to as 

full-length cry1Ab. The Cry1Ab protein is toxic to certain lepidopteran species. 

• The terminator sequence was derived from the nopaline synthase gene of the plant 

pest A. tumefaciens (Entrez Accession Number V00087 (NCBI 2007)). Its 

function is to provide a polyadenylation site 3 (Depicker 1982).

• The right and left border regions of T-DNA from Agrobacterium tumefaciens

nopaline Ti-plasmid.

Transgenic DNA was introduced into the parental cotton line Coker 312, using disarmed 

(non-plant pest causing) Agrobacterium-mediated transformation. Plant cells containing 

the introduced DNA were selected by culturing in the presence of hygromycin. After the 

initial incubation, the broad-spectrum antibiotic cefotaxime was included in the culture 

medium to kill any remaining A. tumefaciens. Because the transformed cells contain 

sequences from plant pathogens, they are explicitly subject to regulation under 7 CFR 

part 340.

 3 Polyadenylation site – facilitates the addition of adenine nucleotides to the 3′ end of messenger 

ribonucleic acid molecules during posttranscriptional modification. 
Page 5 of 16

Data from Southern analyses demonstrate that COT67B plants contain: (1) a single copy 

of cry1Ab (Syngenta 2007, Figure 3-8); (2) a single copy of the actin promoter (Syngenta 

2007, Figure 3-10); (3) no sequences from the transformation plasmid (pNOV4641) 

(Syngenta 2007, Figure 3-12) (i.e.‘backbone sequences’); (4) no sequences from the aph4

gene. Analysis of DNA sequences of COT67B confirmed that the overall integrity of the 

intended insert and the contiguousness of the functional elements have been maintained 

(Syngenta 2007, Appendix 1 - Figure 4). Statistical analyses over multiple generations 

confirm that the cry1Ab gene is stably integrated and is inherited over generations in the 

expected fashion (Syngenta 2007, Figures 3-17 and 3-18). 

Potential of COT67B to Become Invasive and/or a Weed

Cotton is not considered a weed in the U.S. (Holm 1977 and 1997, Muenscher 1980, 

Reed 1977) and is not listed as a Federal noxious weed (USDA-NRCS 2006). APHIS 

assessed whether COT67B is any more likely to become a weed than the nontransgenic 

comparator, or other cultivated cotton. The assessment encompassed a thorough 

consideration of the basic biology of cotton and an evaluation of the unique 

characteristics of COT67B. APHIS analyzed the field test reports and other data included 

in the petition, as well as data from scientific literature on the establishment, 

reproduction, and dispersal processes of cotton as measures of invasiveness and 

weediness. 

Syngenta provided measures of plant growth including; plant height, days to flowering, 

total nodes4

, plant height to node ratio, number of bolls per plant and days to open boll 

(Syngenta 2007, Tables 5-3, 5-8, 5-9, 5-10, 5-16 through 5-19 and Appendix 4). Some 

statistically significant differences in plant growth characteristics were noted in various 

field trials. For instance, plant height was similar to both comparators for early square, 

early bloom and late bloom stage of plant development in 2005 (Syngenta 2007, Tables 

5-8, 5-9 and 5-10) and for early square stage in 2006 (Syngenta 2007, Table 5-16). In 

2006 the early bloom, late bloom, and preharvest stages of COT67B had similar plant 

height when compared to its COT67B null isoline5 (Syngenta 2007, Tables 5-17, 5-18 

and 5-19), however, COT67B on average was slightly shorter than the parental Coker 

312 (Syngenta 2007, Table 5-17, 5-18 and 5-19). The observed variations in growth 

were small, inconsistently seen in all stages of plant development. The differences in 

growth were minor and not found in all growing environments and therefore, not 

expected to be a factor in terms of weediness or pest potential. 

A measure of the reproductive capacity of plants that are propagated by seed, such as 

cotton, is the number of seeds that are produced and the germination and viability of 

 4 A node is the point on the stem from which a leaf grows. 5 Null isoline refers to a plant line that is genetically very similar to another plant line except for the 

presence of a different allele at one locus. Because there is only one copy of the cry1Ab gene in COT67B, 

when the COT67B is crossed with a cultivar that lacks the cry1Ab gene, 50 percent of the progeny will lack 

the cry1Ab gene. In this case, the null isoline refers to the progeny of the original transformed COT67B that 

lack the cry1Ab gene. 
Page 6 of 16

those seeds. Overall, COT67B produced similar or slightly lower levels of seed when 

compared to Coker 312 or the null isoline (Syngenta 2007, Tables 5-4, 5-11 and 5-20). 

Seed germination tests showed no significant differences in relative germination 

(Syngenta 2007, Table 5-12). 

These results on growth characteristics and seed production and germination, indicate 

that the COT67B is not significantly different than its comparators. In addition to the 

results summarized above, APHIS notes that there have been no reports of increased 

weediness associated with other lepidopteran insect resistant cotton lines that have been 

deregulated pursuant to Part 340 and the Plant Protection Act (Carpenter 2002, USDAAPHIS 2009). There is no indication that COT67B possesses a selective advantage that 

would result in increased weediness. Therefore, COT67B lacks the ability to persist as a 

troublesome weed, and there would be no direct impact on current weed management 

practices for cotton cultivation.

Potential for Gene Flow and Gene Introgression from COT67B into its SexuallyCompatible Relatives 

G. hirsutum is a perennial plant that is cultivated as an annual in the U.S. where it is 

grown from Virginia southward and westward to California (McGregor 1976, USDANASS 2009). G. hirsutum is generally self-pollinating and its pollen is not readily 

dispersed by the wind because it is sticky and heavy (Khan 1950, Thies 1953). Insect 

pollination is generally carried out by bumble bees, Melissodes bees, and honey bees 

(McGregor 1976). In assessing the risk of gene introgression from COT67B to its 

sexually compatible relatives, APHIS considered two primary issues: 1) the potential for 

gene flow and introgression, and 2) the potential impact of introgression. 

Although most of the cultivated cotton grown in the U.S. is G. hirsutum, G. barbadense

(Pima cotton) is also grown (USDA-NASS 2009). In addition to these cultivated species,

G. thurberi and G. tomentosum are found in the mountains of southern Arizona and in 

Hawaii, respectively. None of the above are listed (or proposed) as endangered or 

threatened under Federal (USFWS 2009a, USFWS 2009b) or State listings (Arizona 

2009, Hawaii 2001) with the exception of G. hirsutum.

Wild populations of G. hirsutum have been listed as threatened and endangered by the 

State of Florida (Florida 2003). However, in Florida, wild G. hirsutum is not present in 

the northwestern panhandle where cotton cultivation occurs (Coile 2003, USDA-NASS 

2009, Wunderlin 2004). Additionally, because the terms and conditions of EPA’s 

conditional registration for Cry1Ab in cotton prohibits commercial cultivation south of 

Route 60 (near Tampa (EPA 2008)), COT67B is neither expected to be planted 

commercially in the areas of Florida where wild populations of G. hirsutum occur nor 

would they likely be impacted by COT67B planted north of Route 60. Therefore, 

because they are not likely to be present in close proximity, cultivated G. hirsutum is not 

likely to cross with wild populations of G. hirsutum.
Page 7 of 16

G. tomentosum is native to the Hawaiian Islands, occurring primarily in arid, rock, or clay 

coastal plains (Wagner 1999). In laboratory and greenhouse breeding programs with 

hand pollination, G. tomentosum and G. hirsutum are sexually compatible (Meyer 1974), 

form viable progeny and share common pollinators (Pleasants 2010). However, DNA 

marker analyses have not found evidence of genes from G. hirsutum occurring in native 

populations of G. tomentosum (DeJoode 1992). It is possible that the lack of evidence of 

movement of G. hirsutum genes into G. tomentosum is the result of lack of opportunity, 

because cotton has not been grown commercially in Hawaii for at least the last 45 years 

(USDA-NASS 2009). G. tomentosum is not known to be weedy or to have invasive 

characteristics (Holm 1977, Holm 1997, University of Hawaii 2001), and is considered a 

rare plant in Hawaii (University of Hawaii 2001). Because G. tomentosum is not a weedy 

plant, even if cry1Ab did introgress into G. tomentosum it is not likely to become weedy 

or invasive. Finally, EPA’s conditional registration for Cry1Ab prohibits commercial use 

of COT67B in Hawaii (EPA 2008), therefore it would not be planted in Hawaii and the 

probability of outcrossing would not exist. 

The difference in chromosome numbers precludes sexual compatibility of G. hirsutum 

with G. thurberi (OECD 2008). Outcrossing of genes from COT67B to G. thurberi is 

unlikely to occur, because G. thurberi contains only the D genome whereas G. hirsutum 

contains both the A and D genome. In addition, in Arizona, G. thurberi was eradicated 

near cotton growing areas as part of the cotton boll weevil control program (Benson 

1981, Kearney 1960) and the probability of outcrossing would be very unlikely. 

Therefore, USDA has determined that any adverse consequences of gene flow from 

COT67B to wild or weedy species in the U.S. are highly unlikely.

Potential for Transfer of Genetic Information to Organisms with which COT67B 

Cannot Interbreed

Horizontal gene transfer and expression of DNA from a plant species to bacteria is 

unlikely to occur (Keese 2008). First, many bacteria (or parts thereof) that are closely 

associated with plants have been sequenced, including Agrobacterium and Rhizobium 

(Kaneko 2000, Kaneko 2002, Wood 2001). There is no evidence that these organisms 

contain genes derived from plants. Second, 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 (Brown 2003, Koonin 2001). 

Third, transgene DNA promoters and coding sequences are optimized for plant 

expression, not bacterial expression. Thus, even if horizontal gene transfer occurred, 

proteins corresponding to the transgenes are not likely to be produced. Therefore, APHIS 

concludes that horizontal gene transfer is unlikely to occur and thus poses no significant 

environmental or plant pest risk.

Potential for COT67B to have Altered Disease and Pest Susceptibilities

APHIS assessed whether COT67B is likely to have significantly altered disease and pest 

susceptibility. This assessment encompassed a thorough consideration of the introduced 

trait and disease and pest susceptibility data from COT67B field trials. 
Page 8 of 16

Transformation of G. hirsutum with Agrobacterium will not lead to crown gall disease,

because the A. tumefaciens strain GV3101 was disarmed by removing the native T-DNA

(Syngenta 2007, page 31, Klee 2000). (The native T-DNA contains the plant hormone 

genes necessary for the formation of crown gall tumors.) Instead, a T-DNA region that 

contains a cry1Ab gene as well as the regulatory components necessary for their 

expression in the cotton genome, was introduced on plasmid pNOV4641 (Syngenta 2007,

Table 3-1). Further, antibiotics were used to kill any remaining Agrobacterium after the 

transformation. 

The DNA inserted into COT67B includes only, the actin promoter from the plant A. 

thaliana, the cry1Ab gene from the bacterium B. thuringiensis, commonly found in soils,

and the nopaline synthase termination sequence from A. tumefaciens (see the Description 

of the Modification in this document). The A. tumefaciens nopaline synthase terminator 

has been well-characterized and cannot cause disease (Hooykaas 1992). The actin 

promoter sequence is derived from a plant that is not a plant pest and is therefore highly 

unlikely to make COT67B susceptible to disease and plant pests.

COT67B was grown during 2004, 2005, and 2006, in 22 locations encompassing the 

range of geographical and environmental conditions where it is anticipated that it will be 

grown commercially in the U.S. These field trials included two cotton comparators: the 

COT67B null isoline and Coker 312. COT67B plants were examined for changes in the 

types of plant pests that were observed and the relative pest damage that those plants 

incurred (field test reports submitted for COT67B, Syngenta 2007, Tables 1-1 and 5-6). 

When expressed in COT67B, the Cry1Ab protein provides protection from the feeding 

damage incurred by several important lepidopteran pests of cotton (Syngenta 2007,

Supplements 20 and 22).

Data submitted by Syngenta indicate that there are no significant differences for disease 

susceptibility between COT67B cotton and its comparators (field test reports for field 

trials of COT67B submitted to APHIS by Syngenta, Syngenta 2007, Table 5-1). All 

cotton plants (GE and non-GE) produce the anti-nutrients6 gossypol and cyclopropenoid 

fatty acids (OECD 2004). Data presented by Syngenta showed that the levels of both

gossypol and cyclopropenoid produced in COT67B cotton fall within normal ranges of 

commercially available cotton varieties (Syngenta 2007, Table 5-30). Overall, data 

presented by Syngenta indicate that the composition of COT67B is not significantly 

different from its non-transgenic comparators except for the presence of the inserted 

sequence (Syngenta 2007, Tables 5-25 through 5-29 and 5-31). Based upon the field trial 

and composition data, it appears that COT67B exhibits the same pest susceptibility as 

conventional cotton, except for the increased resistance to lepidopteran pests.

 6 Antinutrients are substances that inhibit nutrient metabolism or absorption. Gossypol provides defense 

against herbivory (insect feeding) and microbial attack. Cyclopropenoid fatty acids are known to inhibit the 

desaturation of stearic acid to oleic acid in animals. Because of the toxic effect of these compounds, 

maximum allowable limits have been set for levels of these compounds in cotton that is used for food and 

feed (OECD 2004).
Page 9 of 16

Additionally, it does not appear that COT67B harbors an altered pest or pathogen 

community compared to other cotton varieties (field test reports submitted by Syngenta 

for COT67B, Syngenta 2007, Table 5-1).

Potential Impacts of COT67B on Target and Non-target Organisms, Including 

Beneficial Organisms

For decades, microbial products containing B. thuringiensis (the organism that produces 

the Cry1A protein) have been used to control insect pests on a commercial scale and for 

home garden applications (Glare 2000, OMRI 2008, Shelton 2002). Cry1Ab proteins 

display lepidopteran-specific toxic activity against the early-instar larvae of 

agronomically important insect pests such as pink bollworm (Pectinophora gossypiella), 

tobacco budworm (Heliothis virescens), cotton bollworm (Helicoverpa armigera), 

armyworms (Spodoptera spp.), European corn borer (Ostrinia nubilalis), and 

diamondback moth (Plutella xylostella) (Glare 2000, Van Frankenhuzen 1993, Syngenta 

2007, Supplement 20). 

Plants that were genetically engineered to express the Cry1Ab protein have a history of 

safe use in the U.S. Since the mid-1990s, corn and cotton lines have been 

commercialized without substantiated reports of significant deleterious impacts on nontarget organisms (EPA 2008, Mendelson 2003, OECD 2007, USDA-APHIS 2009). The 

use of transgenic cotton producing the Cry1Ab proteins has been shown to reduce the use 

broad spectrum insecticides

7 without significant impacts on diversity of non-target

insects (Cattaneo 2006, Dively 2005, Marvier 2007, Naranjo 2005, Romeis 2006, Torres 

2005 & 2007, Whitehouse 2005). COT67B is expected to be similar with respect to the 

low potential harm to the environment (Syngenta 2007, Appendix 6, Supplements 12

through 19 of the petition, letter from Syngenta dated July 20, 2007 (which summarizes 

the data from the referenced Supplements), EPA 2008). Because Cry1Ab receptors are

not present in non-target birds and mammals (Hofmann 1988a and 1988b, Shimada 

2006a and 2006b, Van Rie 1989 and 1990), these insecticidal proteins are not expected to 

adversely affect non-target invertebrate and vertebrate organisms (EPA 2008). 

Syngenta submitted data from laboratory and field studies on non-target representative 

species, and other peer reviewed studies that provide evidence for the lack toxicity of 

Cry1Ab (Syngenta 2007, Appendix 6, Supplements 12 through 19, letter from Syngenta 

dated July 20, 2007 (which summarizes the data from the referenced Supplements), EPA 

2008). Assessment of insecticidal transgenic crops include laboratory tests with indicator 

test species to determine potential toxicity at doses of the toxin higher than would be 

anticipated under field conditions (Rose 2007). Selection of representative indicator test 

species was based upon the potential for exposure to Cry1Ab. Syngenta submitted non-

 7 Broad spectrum insecticides are chemical insecticides which kill insects that are causing injury to plants 

and also kill other insects that are not causing injury to the plant. Insects that are inadvertently killed by the 

application of insecticide are called “non-target” insects. Because the Cry1Ab protein is specific for a 

narrow range of insects, use of Cry1Ab to control plant pests is recognized as being benifical to the survival 

of non-target insects (EPA 2008). 
Page 10 of 16

target data for two above-ground arthropods (insidious flower bug (Orius insidious) and 

spotted ladybird beetle (Coleomegilla maculate)), two soil dwelling arthropods (rove 

beetle (Aleochara bilineata) and springtail (Folsomia candida)), a pollinator (honeybee 

(Apis mellifera)), a bird (Bobwhite quail (Colinus virginianus)), a mammal (mouse (Mus 

musculus)), an aquatic invertebrate (water flea (Daphnia magna)), and a fish (catfish 

(Ictalurus punctatus)). The data submitted in the petition indicate that no significant 

adverse effects were observed at the maximum test dose for any of the tested species. 

Other research has also shown no direct adverse effects on insectivores (insects that eat 

insects) in field and laboratory studies with transgenic plants expressing Cry1Ab (Pilcher 

1997, Marvier 2007, Romeis 2004, Romeis 2006). Exposure of aquatic organisms is 

likely very low because cotton pollen is large, sticky, and is not transported long 

distances by the wind (Thies 1953). Seed and plant debris are not expected to be readily 

transported via overland runoff or wind to aquatic habitats.

Based on the above information, APHIS believes that a determination of nonregulated 

status for COT67B will have no adverse effects on non-target organisms. 

Conclusion

APHIS has prepared this plant pest risk assessment in order to determine if event 

COT67B is unlikely to pose a plant pest risk. Based on the information provided by the 

applicant and the lack of plant pest risk from the inserted genetic material, weedy 

characteristics, atypical responses to disease or plant pests in the field, effects on nontargets or beneficial organisms in the agro-ecosystem, and horizontal gene transfer,

APHIS has concluded that event COT67B is unlikely to pose a plant pest risk. 

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