Safety Evaluations of the Cry1Ia1 Protein Found in the Transgenic Potato ‘SpuntaG2’

Quemada, Hector; Zarka, Kelly; Pett, W.; Bothma, Gurling; Felcher, Kimberly J.; Mirendil, Hope; Koch, Muffy; Brink, Johan; Douches, David S.

doi:10.21273/jashs.135.4.325

Cited by 14 publications

(10 citation statements)

References 18 publications

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“…Based on multiple years of field, storage, and agronomic trials, as well as measurement of agronomic traits, it was concluded that 'SpuntaG2' gave complete resistance to potato tuber moth infestation in the field and in storage and that it does not differ from non-transgenic 'Spunta' with respect to yield, other important agronomic properties, tuber quality, and processing traits. Furthermore, two companion articles (Quemada et al, 2010;Zarka et al, 2010) give a complete characterization of 'SpuntaG2' at the molecular level and a thorough protein safety evaluation that demonstrated that 'SpuntaG2' is safe for human consumption. Therefore, in South Africa where potato tuber moth is a serious pest of potatoes, 'SpuntaG2' could provide growers with an alternative to the variable results achieved with approved chemical insecticides.…”

Section: Discussionmentioning

confidence: 99%

“…'SpuntaG2' is a transgenic potato cultivar containing the cry1Ia1 gene from Bacillus thuringiensis for resistance to potato tuber moth. This cultivar has been characterized at the molecular level (Zarka et al, 2010) and has been thoroughly evaluated for safety as a human food source (Quemada et al, 2010). The current study focuses on the agronomic performance of 'SpuntaG2' and its efficacy against potato tuber moth in the field and in storage.…”

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confidence: 99%

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Field and Storage Evaluations of ‘SpuntaG2’ for Resistance to Potato Tuber Moth and Agronomic Performance

Douches¹,

Pett²,

Visser³

et al. 2010

J. Amer. Soc. Hort. Sci.

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‘SpuntaG2’ is a transgenic potato (Solanum tuberosum) cultivar that contains the cry1Ia1 gene for resistance to potato tuber moth (Phthorimaea operculella), which is a serious pest of potato in many parts of the world. Previous studies have characterized ‘SpuntaG2’ at the molecular level and evaluated it for safety as a human food source. The objective here was to determine the efficacy of ‘SpuntaG2’ against the potato tuber moth in the field and in storage and to evaluate its agronomic performance. Efficacy trials at seven South African locations over 5 years indicated that ‘SpuntaG2’ gave complete control of potato tuber moth in the field and storage. The agronomic performance (tuber size and yield) of ‘SpuntaG2’ was not statistically different from ‘Spunta’ or was better than ‘Spunta’ at all locations/years with two exceptions. Three years of trials in Michigan further support these conclusions. Evaluations done during the 2006–07 South African season showed that ‘SpuntaG2’ did not differ from non-transgenic ‘Spunta’ for the following traits: maturity, growth habit, eye morphology, tuber size distribution, tuber shape, skin and flesh color, growth cracks, internal defects, specific gravity, chip color, cooking quality, disease resistance, and pollen fertility. Based on the results of these trials, it was concluded that ‘SpuntaG2’ provides the agronomic performance of ‘Spunta’ with the added benefit of resistance to potato tuber moth.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Field and Storage Evaluations of ‘SpuntaG2’ for Resistance to Potato Tuber Moth and Agronomic Performance

Douches¹,

Pett²,

Visser³

et al. 2010

J. Amer. Soc. Hort. Sci.

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“…Equivalency tests of Cry1Ia1 from GMO potato and E. coli produced Cry1Ia1 showed no difference in size in Western blot. 23 The Western blot results also showed that Cry2Aa2 expressed in tobacco leaves was larger than the E. coli expressed Cry2Aa2. 12 The larger molecular protein on lane 5 could be the cotton Cry1Ac protein precursor.…”

Section: Journal Of Agricultural and Food Chemistrymentioning

confidence: 88%

“…Western blot is widely used across a broad range of biological science disciplines − as well as the detection of GMO proteins expression level, environmental fate or biosafety investigation. ,, Western blot provides useful information, like protein degradation, truncation, and modification after translation, which is not readily gathered from other immunoassays. After gel electrophoretic separation and protein transfer, a target protein is detected by a specific antibody, which confirms the identity of the target protein.…”

Section: Introductionmentioning

confidence: 99%

Development of Monoclonal Antibodies Recognizing Linear Epitope: Illustration by Three Bacillus thuringiensis Crystal Proteins of Genetically Modified Cotton, Maize, and Tobacco

Cao

Zhang

Ning

et al. 2017

J. Agric. Food Chem.

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Bacillus thuringiensis Cry1Ac, Cry1Ia1, and Cry1Ie are δ-endotoxin insecticidal proteins widely implemented in genetically modified organisms (GMO), such as cotton, maize, and potato. Western blot assay integrates electrophoresis separation power and antibody high specificity for monitoring specific exogenous proteins expressed in GMO. Procedures for evoking monoclonal antibody (mAb) for Western blot were poorly documented. In the present study, Cry1Ac partially denatured at 100 °C for 5 min was used as an immunogen to develop mAbs selectively recognizing a linear epitope of Cry1Ac for Western blot. mAb 5E9C6 and 3E6E2 selected with sandwich ELISA strongly recognized the heat semidenatured Cry1Ac. Particularly, 3E6E2 recognized both E. coli and cotton seed expressed Cry1Ac in Western blot. Such strategy of using partially denatured proteins as immunogens and using sandwich ELISA for mAb screening was also successfully demonstrated with production of mAbs against Cry1Ie for Western blot assay in maize.

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“…The research presented here is the first in a series of three articles including Quemada et al (2010) and Douches et al (2010) documenting the extensive research necessary to apply for the release of a GM potato ('SpuntaG2') in South Africa. This project was funded by the U.S. Agency for International Development (Washington, DC) and is a collaborative effort between Michigan State University Potato Breeding and Genetics Program (East Lansing) and the Agricultural Research Council, Vegetable and Ornamental Plant Institute [(ARC-VOPI), Pretoria, South Africa].…”

mentioning

confidence: 99%

Insertion and Characterization of the cry1Ia1 Gene in the Potato Cultivar Spunta for Resistance to Potato Tuber Moth

Zarka¹,

Greyling²,

Gazendam³

et al. 2010

J. Amer. Soc. Hort. Sci.

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Potato tuber moth (Phthorimaea operculella) is a serious pest of potatoes in tropical and subtropical regions of the world, including South Africa. The cry1Ia1 gene (from Bacillus thuringiensis) under the control of the 35S cauliflower mosaic virus promoter was transformed into the potato (Solanum tuberosum) cultivar Spunta to develop a cultivar with resistance to potato tuber moth for release in South Africa. Two transformation events, ‘SpuntaG2’ and ‘SpuntaG3’, were selected and subjected to extensive molecular analyses as required by the regulatory agencies of South Africa. Southern hybridization experiments indicated that ‘SpuntaG2’ and ‘SpuntaG3’ had one and three copies of the cry1Ia1 gene, respectively, and that the gene insertion was stable through multiple clonal generations. Furthermore, the sequence of the cry1Ia1 gene in ‘SpuntaG2’ was compared with the known sequence of the cry1Ia1 gene and found to be identical. Polymerase chain reaction (PCR) amplification using primers for plasmid “backbone” genes demonstrated that ‘SpuntaG2’ contained no backbone plasmid genes, whereas ‘SpuntaG3’ contained several backbone plasmid genes. Therefore, further analyses were limited to ‘SpuntaG2’, and event-specific primers were developed for this cultivar. Analysis of the left and right border regions in ‘SpuntaG2’ demonstrated that the insertion of the cry1Ia1 gene did not disrupt any functional genes nor did it create new open reading frames that encoded proteins with a significant match to the non-redundant sequence database queried by the BLASTP program. Enzyme-linked immunoabsorbent assays (ELISA) tests indicate that the cry1Ia1 gene was expressed at a mean concentration of 2.24 μg·g−1 fresh weight in leaf tissue and 0.12 μg·g−1 fresh weight in tubers. This study demonstrates the extensive molecular characterization that is necessary to apply for deregulation of a genetically modified crop and these data have been used in a regulatory package for the general release of ‘SpuntaG2’.

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Safety Evaluations of the Cry1Ia1 Protein Found in the Transgenic Potato ‘SpuntaG2’

Cited by 14 publications

References 18 publications

Field and Storage Evaluations of ‘SpuntaG2’ for Resistance to Potato Tuber Moth and Agronomic Performance

Field and Storage Evaluations of ‘SpuntaG2’ for Resistance to Potato Tuber Moth and Agronomic Performance

Development of Monoclonal Antibodies Recognizing Linear Epitope: Illustration by Three Bacillus thuringiensis Crystal Proteins of Genetically Modified Cotton, Maize, and Tobacco

Insertion and Characterization of the cry1Ia1 Gene in the Potato Cultivar Spunta for Resistance to Potato Tuber Moth

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