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Living Modified Organism (LMO)
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Decisions on the LMO Risk Assessments  
published: 26 Feb 2019 last updated: 20 Jun 2024
Living Modified Organism identity
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
AAT709A
Yes
AAT-7Ø9AA-4
Cowpea (Vigna unguiculata) was modified through Agrobacterium-mediate transformation to confer resistance to Lepidoptera pests, particularly pod borer (Maruca vitrata). The modified cowpea expresses Bacillus thuringiensis crystal protein Cry1Ab, which has a pore-forming mode of action that selectively damages the midgut epithelium line of feeding larvae, resulting in cell lysis and septicemia and leading to death of the feeding larvae. In addition, the modified cowpea expresses an Escherichia coli neomycin phosphotransferase II cassette, which functioned as a selectable marker for kanamycin resistance during transformation of the cowpea line. 
EN
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.
Cowpea cultivar IT86D-1010
EN
Characteristics of the modification process
pMB4
EN
  • Agrobacterium-mediated DNA transfer
 
0.523 kb
 
 
0.177 kb
 
 
0.190 kb
 
 
0.618 kb
 
 
0.139 kb
 
 
1.726 kb
 
 
1.848 kb
 
 
0.373 kb
 
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
  • BCH-GENE-SCBD-114438-3 SCSV S1 promoter | Subterranean clover stunt virus (SCSV, Subterranean clover stunt virus)
    Promoter
  • BCH-GENE-SCBD-15001-5 Neomycin Phosphotransferase II | Escherichia coli (ECOLX)
    Protein coding sequence | Resistance to antibiotics (Kanamycin)
  • BCH-GENE-SCBD-114274-2 Catalase 1 intron | Ricinus communis (Castor bean)
    Intron
  • BCH-GENE-SCBD-114440-2 SCSV3 terminator | Subterranean clover stunt virus (SCSV, Subterranean clover stunt virus)
    Terminator
  • BCH-GENE-SCBD-103851-5 rbcS Promoter | Arabidopsis thaliana (Thale cress, Mouse-ear cress, Arabidopsis, ARATH)
    Promoter
  • BCH-GENE-SCBD-14985-12 Cry1Ab | Bacillus thuringiensis (Bt, Bacillus, BACTU)
    Protein coding sequence | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • BCH-GENE-SCBD-103853-2 rbcS Terminator | Nicotiana tabacum (Tobacco, TOBAC )
    Terminator
The modified cowpea expresses two gene cassettes: Bacillus thuringiensis delta-endotoxin (crystal) cry1Ab and Escherichia coli neomycin phosphotransferase II (nptII).

Transcription of cry1Ab occurs from the Arabidopsis thaliana ribulose-1,5-bisphosphate carboxylase small subunit promoter and terminates at the Nicotiana tabacum ribulose-1,5-bisphosphate carboxylase small subunit terminator.

Transcription of nptII is controlled by a Subterranean clover stunt virus DNA segment 1 promoter and DNA segment 3 terminator. The coding sequence of nptII is interrupted by the presence of a Ricinus communis catalase 1 intron to prevent leaky expression of the nptII gene cassette. Due to the nature of the promoter, elevated levels of transcription of nptII are expected from this gene cassette.

Note:
  • Vector pMB4 was derived from pART27.
  • The coding sequence of cry1Ab was codon optimized for plant expression. The sequence encodes a 1155-amino acid native Cry1Ab protoxin, which contains a 615-amino acid protein (68.9 kDa) trypsin-resistant, insecticidally active core protein.
  • Southern blot analysis identified a single T-DNA insertion in the genome, comprising of one complete copy of the plasmid pMB4 T-DNA, except for 10 bp and 29 bp truncations at the 5’ and 3’ termini, respectively, and a 735 bp inverted repeat derived from the 3’ end of the T-DNA. 
  • The site of insertion of the T-DNA was mapped to a region on chromosome 2 of the cowpea genome.
  • Event specific PCR analysis indicated that the DNA inserts were stable through at least 10 generations.
EN
LMO characteristics
EN
  • Food
Detection method(s)
EN
Additional Information
EN
Records referencing this document Show in search
Record type Field Record(s)
Country's Decision or any other Communication LMO identification 4
Risk Assessment generated by a regulatory process Living modified organism(s) 5
Living Modified Organism Recipient Organism or Parental Organisms 1
Living Modified Organism Related LMO(s) 1
Laboratory for detection and identification of LMOs LMO(s) detectable by the laboratory 1