Biotechnology Consultation Note to the File BNF No. 000118

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Biotechnology Consultation - Note to the File
Biotechnology Notification File BNF No. 000118

DATE: August 12, 2011


Biotechnology Notification File (BNF) BNF 000118; Cotton Events T304-40 and GHB119


Cotton, Gossypium hirsutum L., Coker 312 variety, Coker 315 variety, event T304-40, event GHB119, T304-40 x GHB119, Lepidopteran insect resistance, Cry1Ab protein, Cry2Ae protein, bar gene, phosphinothricin acetyltransferase, PAT protein, Bayer CropScience LP, TwinLink


This document summarizes our evaluation of biotechnology notification file (BNF) No. 0118 submitted by Bayer CropScience LP (Bayer) on January 12, 2009. The file describes Bayer’s safety and nutritional assessment of bioengineered cotton events T304-40 and GHB119. Bayer subsequently provided additional information in submissions dated July 10, 2009, December 19, 2009, November 2, 2010 and December 7, 2010. FDA evaluated the information in Bayer’s submission to ensure that human food and animal feed regulatory or safety issues, regarding the food or feed derived from the new plant variety have been resolved prior to commercial distribution.

In our evaluation of BNF No. 000118, we considered all information provided by the notifier as well as publicly available information and information in the agency’s files. Here we discuss the outcome of the consultation, but do not intend to restate the information provided in the final consultation in its entirety.

Intended Effect

The intended effect of the modifications in events T340-40 and GHB119 was to confer resistance to common Lepidopteran insect pests of cotton, primarily bollworm and tobacco budworm. To accomplish this objective, Bayer introduced the cry1Ab gene, encoding the Cry1Ab protein, and the bar gene, encoding the phosphinothricin acetyltransferase (PAT) enzyme, used as a selectable marker, into cotton event T340-40 and the cry2Ae gene, encoding the Cry2Ae protein, and the bar gene into cotton event GHB119. The cry1Ab gene was derived from Bacillus thuringiensis (Bt) subsp. berliner, while the cry2Ae gene was derived from B. thuringiensis (Bt) subsp. dakota.

Cotton events T340-40 and GHB119 were subsequently combined through traditional breeding.[1] The cotton line containing both events is hereafter referred to as cotton T304-40 x GHB119. Bayer states that they plan to market this cotton line containing both events.[2] FDA evaluated data provided by Bayer for both the individual cotton events T304-40 and GHB119, as well as the combined T304-40 x GHB119 variety intended for commercial use.

Regulatory Considerations

The Environmental Protection Agency (EPA) defines a plant-incorporated protectant (PIP) as “a pesticidal substance that is intended to be produced and used in a living plant, or in the produce thereof, and the genetic material necessary for production of such a pesticidal substance,” including, “any inert ingredient contained in the plant, or produce thereof” (40 CFR 174.3). EPA regulates PIPs under the Federal Insecticide, Fungicide, and Rodenticide Act and the Federal Food, Drug, and Cosmetic Act. Under EPA regulations, the cry1Ab and cry2Ae genes and resulting expression products are considered pesticidal substances; and the bar gene and resulting expression products are considered inert ingredients.

Genetic Modification and Characterization

The recombinant DNA constructs cry1Ab, cry2Ae and bar in cotton events T304-40 and GHB119 fall under the regulatory purview of EPA.

Protein Characterization

The expression products of the recombinant DNA constructs Cry1Ab, Cry2Ae, and bar in cotton events T304-40 and GHB119 fall under the regulatory purview of EPA.

Food & Feed Use

Cotton (Gossypium hirsutum L.) is grown worldwide primarily as a source of fiber for the textile industry. Cottonseed-derived products are used in human food, animal feed, and a range of industrial products. Food uses include cottonseed oil and cotton linters. Cottonseed oil is highly refined to remove naturally occurring toxicants, gossypol and cyclopropenoid fatty acids (CPFAs). Cottonseed oil is primarily consumed as a salad or cooking oil. Cotton linters are short fibers that remain on cotton seeds after ginning. They are removed from the seeds and processed into pure cellulose, which is used, for example, in casings for bologna, sausages, and frankfurters, and in other products such as ice cream and salad dressings.

Whole cottonseed, cottonseed meal, crude cottonseed oil, hulls, and cotton gin trash may be used in animal feeds for cattle, sheep, goats, horses, poultry, swine, fish and shrimp. Small amounts of cotton linters have been used in animal feeds. Whole cottonseed has been used as a source of protein and energy in cattle. Cottonseed meal is the product obtained from the flakes or cake after oil removal and is used as a protein supplement in animal feed. Cottonseed hulls and gin trash are used as a source of fiber in animal feeds.


Scope of Analysis

Bayer analyzed the composition of ginned cottonseed (fuzzy seed) derived from events T304-40 and GHB119 and cotton T304-40 x GHB119. These transgenic crops were compared to non-transgenic cotton control varieties grown at the same locations, Coker 315, Coker 312, and Coker 315 respectively. Bayer also provided information on processed fractions (such as linters, cottonseed meal, hulls, and refined bleached deodorized oil) from cotton T304-40 x GHB119.

Study Design - Compositional Analyses

Bayer analyzed ginned cottonseed containing individual events T304-40 and GHB119 as well as cotton T304-40 x GHB119 containing both events. Bayer provided analytical data for the following components:

  • Proximates: moisture, crude protein, crude fat, ash, and carbohydrates calculated by difference
  • Neutral detergent fiber (NDF) and acid detergent fiber (ADF)
  • Amino acids (18)
  • Fatty acids (reported data on 9 major fatty acids for individual events and 12 fatty acids for cotton T304-40 x GHB119)
  • Minerals (calcium, iron, magnesium, phosphorus, potassium, and zinc)
  • Vitamin E (alpha tocopherol and/or total tocopherol)
  • Antinutrients and potential toxicants (phytic acid, total gossypol, free gossypol, sterculic acid, malvalic acid, and dihydrosterculic acid)

Bayer provided analytical results for transgenic cottonseed T304-40, GHB119 and T304-40 x GHB119 collected from plants that were sprayed and not sprayed with glufosinate. The non-transgenic control cotton is sensitive to glufosinate and was not sprayed. There were eight field sites for event T304-40, six field sites for event GHB119, and seven field sites for cotton T304-40 x GHB119. At each of these field sites, seed was planted in a randomized block design with three replicates per block. The analytical results are presented on a dry weight basis except for fatty acids and cyclopropenoid fatty acids which are provided as percentages of total fatty acids.

Results of analyses:
Events T304-40 and GHB119

Bayer stated that the initial transformants containing events T304-40 and GHB 119 were self-pollinated for five generations prior to obtaining seed for compositional analysis. Statistical assessments of the compositional data were conducted using mixed model analysis of variance across all sites, referred to as the over-all sites analysis. The model included treatment, site and treatment by site with treatment and site being considered fixed factors. For some components, the over-all sites analysis was not valid since significant treatment x site interaction was detected. For these components, a by-site analysis was conducted.

Although Bayer observed that there were statistically significant differences in the mean levels of several components between the transgenic line and control samples, these differences were small (usually within one standard deviation). Furthermore, the mean values for each of these components were comparable to the values in the published scientific literature. Bayer concluded that the minor differences in specific components between the transgenic line and its respective comparator for events T304-40 and GHB119 were not biologically meaningful.

Cotton T304-40 x GHB119

Bayer provided composition data on cotton T304-40 x GHB119 containing both individual events combined. Bayer analyzed in total 63 samples of cotton T304-40 x GHB119 (cotton T304-40 x GHB119 unsprayed, cotton T304-40 x GHB119 sprayed, and the comparator line Coker 315 from 3 blocks at 7 locations) for 50 components. The results were analyzed with the analysis of variance methods. The statistical evaluation of data collected from all sites (over-all sites analysis) was not valid for some components due to significant site x treatment interactions. The analytical results for these compounds were analyzed on a by-site basis. Bayer stated that statistical differences at the 5 percent level (P<0.05) were declared to be significant.

Bayer reported that in both the over-all sites analysis and by-site analysis statistically significant differences were observed in the mean levels of several components between cotton T304-40 x GBH119 and the non-transgenic control cotton. Bayer noted that these differences were small and that the mean levels of these components were within the reference ranges for commercial cotton seeds. Bayer concluded that cotton T304-40 x GBH119 is indistinguishable from the non-transgenic cotton with the exception of intentionally added two Bt proteins and the PAT protein.

Bayer also analyzed processed fractions from cotton T304-40 x GHB 119, including fuzzy seed, lint, linters, delinted cottonseed, defatted cottonseed meal (toasted and untoasted), hulls, and oil (crude and refined). Bayer concluded that the levels of all components measured in the processed products derived from cotton T304-40 x GHB119 were comparable with those measured in the processed products derived from non-transgenic control cotton.

Additional Studies

Bayer reported the results of a 42-day broiler chicken feeding study comparing birds fed a diet of 10% toasted cottonseed meal from cotton T304-40 x GHB119, cottonseed meal from Coker 312, and cottonseed meal from a non-transgenic commercial cotton variety. Bayer stated that no treatment-related differences were observed between the chickens receiving cotton T304-40 x GHB119, the comparator, and the commercial variety.


FDA evaluated Bayer’s submission to determine whether the developer’s product raises any safety or regulatory issues with respect to the intended modification or with respect to the food itself. Based on the information provided by the company and other information available to the agency, FDA did not identify any safety or regulatory issues under the Federal Food, Drug and Cosmetic Act or current FDA regulations that would require further evaluation at this time.

Bayer has concluded that cotton derived from events T304-40 and GHB119 and the foods and feeds obtained from these events are as safe as conventional cotton varieties and with the exception of the plant-incorporated protectant proteins, are not materially different in composition or any other relevant parameter from other cotton varieties now grown, marketed, and consumed in the U.S. At this time, based on Bayer’s data and information, the agency considers Bayer’s consultation on cotton derived from events T304-40 and GHB119 to be complete.

Richard E. Bonnette

[1]Bayer states that to obtain cotton T304-40 x GHB119 the initial transformants containing events T304-40 and GHB119 were each crossed with a conventional cotton breeding line and screened for resistance to glufosinate. The F1 glufosinate-resistant plants were subsequently backcrossed into the conventional line. At the BC2F1 generation, the lines were crossed to produce cotton T304-40 x GHB119.

[2]Bayer declares that the company does not intend to release cotton containing individual events for commercial use nor does the company intend to derive commercial varieties from these events beyond the cotton containing both events.

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