Biotechnology Consultation Note to the File BNF No. 000119
Return to inventory: Submissions on Bioengineered New Plant Varieties
Biotechnology Consultation - Note to File
Biotechnology Notification File No. 0119
August 5, 2010
MON 87701 Lepidopteran insect-resistant soybean
Soybean; Glycine max; insect resistance; Lepidopteran resistance; MON 87701; OECD unique identifier MON-877Ø1-2, Cry1Ac protein from Bacillus thuringiensis subsp. kurstaki; cry1Ac gene; The Monsanto Company
This document summarizes our evaluation of biotechnology notification file (BNF) No. 0119. In a submission dated May 28, 2009, the Monsanto Company (Monsanto) submitted a safety and nutritional assessment of MON 87701. Monsanto provided additional information in a submission dated December 16, 2009. Monsanto is voluntarily consulting with the agency as discussed in the agency's 1997 Guidance on Consultation Procedures for Foods Derived from New Plant Varieties. Here we describe the outcome of the consultation but do not recapitulate the entirety of the information we considered.
The intended effect of the modification in MON 87701 is to confer resistance to targeted Lepidopteran insect pests. To accomplish this objective, Monsanto introduced the cry1Ac gene from Bacillus thuringiensis (Bt) subsp. kurstaki. The cry1Ac gene encodes the Cry1Ac protein.
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 the produce thereof, and the genetic material necessary for the 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 cry1Ac gene and resulting expression products in the MON 87701 soybean line are considered pesticidal substances.
Genetic Modification and Characterization
The recombinant DNA construct containing the cry1Ac gene in the MON 87701 soybean line falls under the regulatory purview of EPA.
The expression products of the recombinant DNA construct containing the cry1Ac gene in the MON 87701 soybean line fall under the regulatory purview of EPA.
Food & Feed Use
Soybean (Glycine max) is grown around the world for a variety of food, feed, and industrial uses. Soybean seeds are primarily processed into oil and meal. Soybean oil is rich in polyunsaturated fatty acids and is commonly used as a salad and cooking oil and in the production of margarine and other food ingredients. A small fraction of soybean meal is further processed into soy flours and soy proteins for a variety of food uses. Traditional foods prepared from soybeans include tofu, miso, soymilk, tempeh, and soy sauce.
The preponderance of soybean meal is used in animal feed, primarily in poultry, swine, and beef and dairy cattle diets. Soybean meal is processed in moist heat to inactivate trypsin inhibitors and lectins, which are antinutrients occurring in raw soybeans.
Scope of Analysis
Monsanto analyzed the composition of forage and seed from the MON 87701 soybean and compared it to a non-transgenic soybean control variety, A5547 (control), which has a genetic background similar to MON 87701. Monsanto also evaluated the composition of forage and seed from a total of twenty commercial non-transgenic soybean varieties ("reference varieties") grown under the same field conditions as MON 87701 and control soybeans. Monsanto used the data derived from the reference varieties to generate a 99% tolerance1 interval for each analyte. Monsanto states that these data illustrate the natural variability in commercially grown soybean varieties grown under similar field conditions. The compositional analysis included key nutrients and antinutrients.
Study Design - Compositional Analyses
Monsanto conducted compositional analyses on seed and forage from transgenic (MON 87701), control and reference varieties grown at five field sites, with three replicates per site in a randomized complete block design, in soybean-growing areas of the United States. Monsanto measured and evaluated 57 components in seed and seven in forage. Monsanto analyzed forage for proximates (crude protein, crude fat, moisture, ash), carbohydrates by calculation, acid detergent fiber (ADF), and neutral detergent fiber (NDF). Compositional analysis of seed included measurement of the proximates (crude protein, crude fat, moisture, ash), carbohydrates by calculation, ADF, NDF, fatty acids (C8-C22), 18 amino acids, Vitamin E, isoflavones (daidzein, genistein, and glycitein), and antinutrients (phytic acid, trypsin inhibitor, lectin, raffinose, and stachyose).
Monsanto applied a mixed model analysis of variance for mean values calculated from data aggregated from a combined-site analysis, and also conducted five individual site analyses. The compositional data obtained for MON 87701 soybean forage and seed was evaluated for biological relevance against the 99% tolerance interval derived from analysis of the reference varieties, data from published scientific literature and publicly available soybean crop composition databases such as the International Life Sciences Institute Crop Composition Database (ILSI-CCD).2 Monsanto reported the composition data by providing mean values, ranges, p-values, confidence levels and tolerance intervals for the transgenic and control samples. Monsanto used a p-value of ≤0.05 to identify a significant statistical difference between the conventional control and MON 87701 soybeans for analytes from both seed and forage.
Results of analyses:
Compositional analysis of soybean forage
Monsanto reported no significant statistical difference (p≥0.05) between MON87701 and A5547 for the seven components analyzed in the combined-site analysis for forage. Furthermore, all of the values for MON87701 and A5547 were within the 99% tolerance interval of the reference values and comparable to that published in the scientific literature and publicly available databases (ILSI-CCD). Monsanto concluded that forage from MON 87701 is not materially different from the soybean control.
Compositional analysis of soybean seed
Monsanto noted that nine fatty acids in harvested seed had more than 50% of the observations below the assay limit of quantitation and, as a result, were excluded from the statistical analysis. Monsanto reported significant statistical difference (p≤0.05) for 15 of the 48 components analyzed in the combined-site analysis for seed. These components included crude protein, nine amino acids (alanine, glycine, histidine, isoleucine, leucine, lysine, serine, threonine and valine), carbohydrates, one fatty acid (22:0 behenic acid), vitamin E, and two antinutrients (daidzein and trypsin inhibitor). Mean levels of crude protein from MON 87701 were statistically different from control and reflected the higher levels of the nine amino acids mentioned above. In particular, the level of histidine was shown to be higher than the control at two different test sites. Monsanto also measured higher levels of vitamin E in MON 87701 compared to conventional soybean control in four of the five individual sites. Monsanto noted that the magnitude of differences in these measurements were small, and not consistent in more than two sites. The mean values of all these components were within the calculated 99% tolerance interval of the reference varieties grown concurrently at the same sites and comparable to values in published scientific literature and publicly available databases (ILSI-CCD). Based on the data, Monsanto considers that the measured values for MON 87701 were within the natural variability of this crop, and the differences were not biologically meaningful.
Monsanto's compositional analysis of MON 87701 seeds also identified statistically significant differences for trypsin inhibitor and daidzein. In each case, the differences were not reproducible across sites and the values for MON 87701 and the control variety were within the 99% tolerance interval established for commercial reference varieties, and comparable to values reported in ILSI-CCD. Monsanto concluded that MON 87701 was compositionally equivalent to conventional soybean and that these observed differences were small, inconsistent, and not biologically significant, reflecting natural variation in soybeans.
Monsanto compared the IgE-binding capacity of endogenous soybean allergens in both MON 87701 and a control variety to commercial soybean reference varieties, using human sera from both soybean-allergic and non-allergic subjects. All results were within the 99% tolerance interval calculated for each subject's serum with respect to the commercial reference varieties. Monsanto concluded that IgE binding to soybean allergens in MON 87701 is comparable to IgE binding in commercially available soybean varieties.
Monsanto voluntarily consulted with the agency as discussed in the agency's 1997 Guidance on Consultation Procedures for Foods Derived from New Plant Varieties. Monsanto has concluded that its soybean variety, MON 87701 soybean, and the foods and feeds derived from it are as safe as and are not materially different in composition or any other relevant parameter from other soybean varieties. At this time, based on information provided by Monsanto, the agency considers Monsanto’s consultation on MON 87701 soybean to be complete.
1A 99% tolerance interval represents, with 95% confidence, 99% of the values contained in the population of commercial conventional soybean varieties.
2Monsanto used version 3.0 (accessed on 8-27-06) of the ILSI Crop Composition Database in its analysis. The database is maintained by ILSI and can be accessed at http://www.cropcomposition.org/.