DD2008-71: Determination of the safety of Monsanto Canada Inc.'s Delicious Soybean

Issued: 2008-03

This decision document has been prepared to explain the regulatory decision reached under Directive 95-03, entitled "Guidelines for the Assessment of Novel Feeds: Plant Sources".

The Canadian Food Inspection Agency (CFIA), specifically the Animal Feed Division of the Animal Products Directorate, has evaluated information submitted by Monsanto Canada Inc. regarding Delicious Soybean, which has reduced glycinin content. The CFIA has determined that this novel feed does not present livestock feed safety concerns when compared to currently commercialized soybean varieties in Canada.

Taking into account this evaluation, use as livestock feed of Delicious Soybean is therefore authorized by the Animal Feed Division of the Animal Products Directorate as of March 11, 2008. Any soybean lines derived from Delicious Soybean may also be used as livestock feed, provided that (i) no inter-specific crosses are performed, (ii) the intended uses are similar and (iii) it is known based on characterization, that these plants do not display any additional novel traits and are substantially equivalent to currently grown soybean varieties in Canada, in terms of their potential environmental impact and livestock feed safety.

Delicious Soybean is subject to the same phytosanitary import requirements as its unmodified counterpart.

Table of contents

1. Brief identification of the modified plant
2. Background information
3. Description of the novel trait
   1. Reduced glycinin content
   2. Development method
   3. Stable expression
4. Criteria for the environmental assessment
5. Criteria for the livestock feed assessment
   1. Potential impact of delicious soybean on livestock nutrition
   2. Potential impact of delicious soybean on livestock and workers or bystanders
6. New information requirements
7. Regulatory decision

I. Brief identification of the modified plant

Designation of the modified plant: Delicious Soybean

Applicant: Monsanto Canada Inc.

Plant species: Soybean (Glycine max L.) Merr)

Novel traits: Reduced glycinin content

Trait introduction method: Conventional breeding

Proposed use of the modified plant: Commercial production of soybeans in an identity preserved system for human consumption. These plants are not intended to be grown outside the normal cultivation area for soybeans in Canada.

II. Background information

Monsanto Company has developed a soybean line, designated as Delicious Soybean, which has reduced content of the glycinin protein fraction (11S globulin). This trait is intended to impart physical properties that can be utilized to develop soy-based food ingredients with "favourable" taste and texture characteristics.

Glycinin, the 11S globulin protein, is one of the major soybean storage proteins and is composed of six subunits. The development of Delicious Soybean resulted from traditional crossing of a line with low levels of glycinin (lacking the glycinin subunits A1aB2, A1bB1b, A2B1 and A5A4B3) developed through conventional breeding methods, with a wild-type soybean (Glycine soja) naturally lacking the A3B4 subunit. Thus the storage protein profile of Delicious Soybean lacks five glycinin subunits.

Monsanto has provided data on the identity of Delicious Soybean, a detailed description of the development method and breeding history as well as information about the genes involved in the reduced glycinin trait and the stability of the expression of this trait.

Delicious Soybean has been field tested in the United States and compositional data for the trial years 2004 and 2005 were submitted along with agronomic data from 2006. Some of the locations of these trials share similar environmental and agronomic conditions to soy production areas in Southwestern Ontario and were considered representative of major Canadian soybean growing regions.

Agronomic characteristics of Delicious Soybean such as seed emergence, plant height, lodging, shattering, maturity, moisture and yield were provided and were shown to be similar to commercial soybean lines.

Nutritional components of Delicious Soybean, such as proximates, amino acids and fatty acids were compared with those of commercial reference varieties and found to be similar.

The Plant Biosafety Office of the Plant Products Directorate, CFIA, with advice from the Biotechnology Environmental Release Assessment (BERA) Unit of the Science Strategies Division, CFIA, has reviewed information provided by Monsanto Canada Inc. and agrees with Monsanto Canada Inc.'s determination, based on this information, that Delicious Soybean is not a Plant with a Novel Trait (PNT) and therefore does not require an assessment of environmental safety.

The Animal Feed Division of the Animal Products Directorate, CFIA, has reviewed the submitted data to determine the safety and efficacy of Delicious Soybean or its derivatives as a livestock feed, as described in Directive 95-03, entitled "Guidelines for the Assessment of Novel Feeds: Plant Sources". The Animal Feed Division has considered:

• potential impact of Delicious Soybean on livestock nutrition; and
• potential impact of Delicious Soybean on livestock and workers or bystanders.

Monsanto Canada Inc. has provided the CFIA with a method for the detection and identification of Delicious Soybean.

III. Description of the novel trait

1. Reduced glycinin content

Glycinin, one of the major storage proteins in soybeans, is a hexameric protein with a molecular weight of approximately 350 kDa with five different subunits: G1, G2, G3, G4, and G5. Various combinations of each subunit are assembled into a trimer and covalently stabilized through disulfide bond linkages; two trimers then form the final hexameric protein. Glycinin typically accounts for around 35% of total protein in soybean seed however selective breeding can reduce these levels, as in the case of Delicious Soybean.

The two major storage proteins in soybeans are glycinin and beta-conglycinin, each with different physical, chemical and functional properties. With respect to the functional properties of these proteins, there are notable differences in heat stability, solubility in the presence of metal ions (particularly calcium), binding constants for off-flavor compounds and gel strength. The use of soybeans with reduced glycinin content in food products is intended to improve food processing qualities and impact mouth feel, water retention (texture), and flavour/taste profiles.

2. Development method

The development of Delicious Soybean resulted from crossing a line with low levels of glycinin (lacking the glycinin subunits A1aB2, A1bB1b, A2B1 and A5A4B3) developed through conventional breeding methods, with a wild-type soybean (Glycine soja) naturally lacking the A3B4 subunit. From resulting progeny, one line (B2G2), was identified which lacked all 5 subunits. The storage protein composition was monitored using molecular makers which were verified by SDS-PAGE. Additionally, agronomic traits were used as selection criteria, following typical field breeding practices over multiple years and locations.

3. Stable expression

The traits affecting the composition of interest exhibit normal Mendelian segregation and can be selected through the use of molecular markers. Chi square analysis demonstrated that the genes of interest fit the expected segregation ratio normally exhibited by single recessive genes with Mendelian inheritance patterns. The stability of the traits was demonstrated over three generations.

IV. Criteria for the environmental assessment

The Plant Biosafety Office of the Plant Products Directorate, CFIA, with advice from the Biotechnology Environmental Release Assessment (BERA) Unit of the Science Strategies Division, CFIA, has reviewed information provided by Monsanto Canada Inc. and agrees with Monsanto Canada Inc.'s determination, based on this information, that Delicious Soybean is not a PNT and therefore does not require an assessment of environmental safety.

V. Criteria for the livestock feed assessment

1. Potential impact of delicious soybean on livestock nutrition

Nutritional composition

The compositional equivalence of Delicious Soybean to twelve commercial soybean varieties was determined using data generated from eleven field sites in the United States during the 2004 and 2005 growing seasons. Seed samples were analyzed for proximates, acid detergent fibre (ADF), neutral detergent fibre (NDF), fatty acids, amino acids, vitamin E and isoflavones (diadzein, genistein and glycitein). Crude protein, crude fat, ash, ADF and NDF means measured in Delicious Soybean were within ranges seen in the commercial soybean varieties and ranges found in the Agriculture and Food Systems Institute (AFSI) database for soybeans. The means for palmitoleic, stearic, oleic, linoleic, arachidic, eicosenoic, and behenic acids in Delicious Soybean were within ranges seen in the commercial soybean varieties. In 2005 palmitic and linolenic acids means in Delicious Soybean were outside of the ranges seen in the commercial soybean varieties, but were within ranges found in the AFSI database and/or literature sources. Amino acids means measured in Delicious Soybean in 2004 and 2005 were within the ranges of the commercial soybeans or AFSI database except for arginine, histidine and lysine values measured in 2005. However the means of these amino acids were within ranges found in other literature sources. The vitamin E mean measured in Delicious Soybean was within the range of the commercial soybean varieties and the AFSI database. The means of genistein and glycitein were within the ranges seen in the commercial soybean varieties. The diadzein mean for Delicious Soybean was outside of the range seen in the commercial soybean varieties, but was within the range found in the AFSI database for soybeans.

Antinutritional factors

Phytic acid, trypsin inhibitor, lectin, raffinose and stachyose were analyzed in Delicious Soybean seed and compared to commercial reference soybean varieties. The means measured in Delicious Soybean for these analytes fell within the range of values measured for the commercial reference varieties, the AFSI database, or cited literature sources.

The evidence provided by Monsanto Canada Inc. supports the conclusion that the nutritional composition of Delicious Soybean is substantially equivalent to commercial soybean varieties.

2. Potential impact of delicious soybean on livestock and workers or bystanders

Delicious Soybean was not modified to express a new protein, or a modified form of an endogenous protein, but rather to reduce the expression of the storage protein glycinin. As such, this modification would not be expected to result in the expression of a novel allergen or toxin or the altered expression of endogenous toxins and allergens.

Based on the characterization data provided (nutritional composition and agronomic data), it is unlikely that the reduced glycinin trait has had any unintended effects on the modified plant.

The evidence provided by Monsanto Canada Inc. supports the conclusion that Delicious Soybean will not result in altered impacts on livestock and workers/by-standers compared to current commercial soybean lines.

VI. New information requirements

If at any time, Monsanto Canada Inc. becomes aware of any information regarding risk to the environment, including risk to human or animal health, which could result from the release of Delicious Soybean materials in Canada or elsewhere, Monsanto Canada Inc. will immediately provide such information to the CFIA. On the basis of such new information, the CFIA will re-evaluate the potential impact of Delicious Soybean on the environment, livestock and human health, and may re-evaluate its decision with respect to the livestock feed use and environmental release authorizations of Delicious Soybean.

VII. Regulatory decision

Based on the review of submitted data and information by Monsanto Canada Inc., including comparisons of Delicious Soybean with its unmodified soybean counterparts, the Animal Feed Division of the Animal Products Directorate has concluded that Delicious Soybeans do not confer any characteristic that would raise any concerns regarding the feed safety of Delicious Soybean. Soybean and several of its byproducts are currently listed in Schedule IV of the Feeds Regulations and are, therefore, approved for use in livestock feeds in Canada. Delicious Soybean has been assessed and found to be as safe as and as nutritious as traditional soybean varieties. Delicious Soybean and its products are considered to meet present ingredient definitions and are approved for use as livestock feed ingredients in Canada.

Taking into account this evaluation, livestock feed use of Delicious Soybean is therefore authorized by the Animal Feed Division of the Animal Products Directorate as of March 11, 2008. Any soybean lines derived from Delicious Soybean may also be used as livestock feed, provided no inter-specific crosses are performed, provided the intended uses are similar, and provided it is known based on characterization that these plants do not display any additional novel traits and are substantially equivalent to currently grown soybean in Canada, in terms of their potential environmental impact and livestock feed safety.

Delicious Soybean is subject to the same phyto-sanitary import requirements as its unmodified counterparts.

This bulletin is published by the Canadian Food Inspection Agency. For further information, please contact the Animal Feed Division at:

Animal Feed Division
Animal Products Directorate
59 Camelot Drive
Ottawa, Ontario K1A 0Y9
Telephone: 613-225-2342
Fascimile: 613-773-7565