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Living Modified Organism
(LMO)
The image below identifies the LMO through its unique identifier, trade name and a link to this page of the BCH. Click on it to download a larger image on your computer. For help on how to use it go to the LMO quick-links page.
Pod borer resistant cowpea
EN
DPS200V2
Yes
DPS-2ØØV2-2
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Organization:Donald Danforth Plant Science Center ()Non-governmental organization (NGO)975 N Warson Rd.St. Louis, Missouri
63132, United States of AmericaPhone: +1 314.587.1000, +1 314.406.4287,Fax:Email: kroeber@danforthcenter.org,Website: https://www.danforthcenter.org/,
Genetically modified cowpea (Vigna unguiculata) event, DPS200V2, expresses Bacillus thuringiensis Cry1Bb1 that confers resistance resistance to Lepidoptera pests, particularly maruca pod borer (Maruca vitrata). The crystal delta-endotoxin has a similar mode of action to other crystal proteins, binding receptors in the epithelial gut lining of feeding larvae to form pores, which then allows for an influx of water, cell swelling and eventual lysis. This leads to the destruction of the larval midgut epithelium and death of susceptible larvae.
The term “Recipient organism” refers to an organism (either already modified or non-modified) that was subjected to genetic modification, whereas “Parental organisms” refers to those that were involved in cross breeding or cell fusion.
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BCH-ORGA-SCBD-103617-3 Organism Vigna unguiculata (Cowpea, Black eyed pea)Crops
EN
pDIICI-200
EN
- Agrobacterium-mediated DNA transfer
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
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BCH-GENE-SCBD-272521-2 Cry1Bb | Bacillus thuringiensis (Bt, Bacillus, BACTU)Protein coding sequence | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths), Cotton bollworm (Helicoverpa spp.), European corn borer (Ostrinia nubilalis), Fall armyworm (Spodoptera frugiperda))
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BCH-GENE-SCBD-115757-2 Actin 2 terminator | Arabidopsis thaliana (Thale cress, Mouse-ear cress, Arabidopsis, ARATH)Terminator
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BCH-GENE-SCBD-104802-5 Polyubiquitin10 gene promoter | Arabidopsis thaliana (Thale cress, Mouse-ear cress, Arabidopsis, ARATH)Promoter
The modified cowpea expresses a Bacillus thuringiensis crystal protein (cry1Bb1) gene cassette. The coding sequence is under control of an Arabidopsis thaliana ubiquitin 10 promoter and actin 2 terminator. Due to the constitutive nature of the promoter, high levels of transcription are expected in all plant tissues.
Note:
Note:
- The cowpea was initially transformed with T-DNA containing an additional three genetic cassettes. However, during the development of the cowpea line, the marker cassettes were excised during germline-directed Cre-lox recombination:
- A. thaliana APETALA1 promoter (1.894 kb) – Bacteriophage P1 recombinase cre (1.221 kb) – Solanum tuberosum proteinase inhibitor II terminator (0.311 kb);
- Glycine max ubiquitin promoter + intron (1.946 kb) – synthetic chloroplast transit peptide (0.165 kb) – Streptomyces spectabilis spectinomycin phosphotransferase (spcN; 0.996 kb) – A. thaliana ubiquitin 14 terminator (0.902 kb); and
- Glycine max elongation factor 1 alpha promoter + intron (1.501 kb) – Discosoma sp. DsRed2 (0.678 kb) – A. thaliana ubiquitin 3 terminator (1.088 kb).
- The excision occurs during early plant development. For more information regarding germline-directed Cre-lox recombination, kindly refer to Van Ex and others (2009) and Verweire and others (2007).
EN
- Feed
- Food
EN
Development of the transformation vector:
A derivative of the pCAMBIA2300 binary vector, p5000, was modified by removing the Escherichia coli neomycin phosphotransferase II plant-selectable marker expression cassette and replacing it with the Cre recombinase gene driven by Arabidopsis floral homeotic gene APETALA1, and the Streptomyces spectabilis spectinomycin resistance selectable marker cassette. The gene encoding the Discosoma sp. red fluorescent protein (DsRed2) was also included for visual identification of transformed plants. In addition, the base vector was designed to include restriction endonuclease sites and sequences to facilitate Golden Gate cloning of the gene-of-interest expression cassette. Plasmid pDIICI-200 was introduced into Agrobacterium tumefaciens strain LBA4404 and used to transform cowpea embryonic axis explants using established protocols (see Che and others (2021)).
A derivative of the pCAMBIA2300 binary vector, p5000, was modified by removing the Escherichia coli neomycin phosphotransferase II plant-selectable marker expression cassette and replacing it with the Cre recombinase gene driven by Arabidopsis floral homeotic gene APETALA1, and the Streptomyces spectabilis spectinomycin resistance selectable marker cassette. The gene encoding the Discosoma sp. red fluorescent protein (DsRed2) was also included for visual identification of transformed plants. In addition, the base vector was designed to include restriction endonuclease sites and sequences to facilitate Golden Gate cloning of the gene-of-interest expression cassette. Plasmid pDIICI-200 was introduced into Agrobacterium tumefaciens strain LBA4404 and used to transform cowpea embryonic axis explants using established protocols (see Che and others (2021)).
| Record type | Field | Record(s) | |
|---|---|---|---|
| Risk Assessment generated by a regulatory process | Living modified organism(s) | 2 | |
| Country's Decision or any other Communication | LMO identification | 2 | |
