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Modified Organism
Cowpea resistant to Lepepidoptera pests
Record information and status
Record ID
114508
Status
Published
Date of creation
2019-03-11 16:06 UTC (austein.mcloughlin@cbd.int)
Date of publication
2019-03-11 16:06 UTC (austein.mcloughlin@cbd.int)

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.

LMO name
Cowpea resistant to Lepepidoptera pests
Transformation event
-
Developer(s)
AATF
The African Agricultural Technology Foundation (AATF)
P.O. Box 30709
Nairobi
Kenya, 00100
Phone:+ 254-20 422 3700
Email:aatf@aatf-africa.org
Url:The African Agricultural Technology Foundation
Description
Cowpea (Vigna unguiculata) was modified to express both crystal proteins Cry1Ab and Cry2Ab from Bacillus thuringiensis for resistance to Lepidoptera pests, particularly Maruca vitrata (pod borer). The Cry2Ab protein functions as a backup for Cry1Ab.
Recipient Organism or Parental Organisms
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.
Vigna unguiculata - Cowpea, Black eyed pea
Related LMOs
Cowpea resistant to lepidoptera pests
AATF Resistance to antibiotics - Kanamycin Resistance to diseases and pests - Insects - Lepidoptera (butterflies and moths)
AAT-7Ø9AA-4 - Pod Borer-resistant cowpea
AATF and Institute of Agricultural Research Resistance to antibiotics - Kanamycin Resistance to diseases and pests - Insects - Lepidoptera (butterflies and moths)
Show detection method(s)
Characteristics of the transformation process
Vector
pMB4 (derived from pART27)and pMB6
Techniques used for the modification
  • Agrobacterium-mediated DNA transfer
Genetic elements construct
 
rbcS Promoter
1.73 Kb
 
 
Cry1Ab
1.85 Kb
 
 
rbcS Terminator
0.37 Kb
 
 
SCSV1 promoter
0.52 Kb
 
 
Neomycin Phosphotransferase II
0.18 Kb
 
 
Catalase 1 intron
0.19 Kb
 
 
Neomycin Phosphotransferase II
0.62 Kb
 
 
pMB4 intervening sequence
0.18 Kb
 
 
SCSV3 terminator
0.14 Kb
 
 
SCSV1 promoter
0.53 Kb
 
 
Neomycin Phosphotransferase II
1.16 Kb
 
 
SCSV3 terminator
0.14 Kb
 
 
rbcS Promoter
1.73 Kb
 
 
Transit peptide and first intron of Rubisco SSU
0.27 Kb
 
 
Cry2Ab2
1.91 Kb
 
 
rbcS Terminator
0.46 Kb
 
Further details
Notes regarding the genetic elements introduced or modified in this LMO
DNA insertion from pMB4:
Transcription of cry1Ab occurs from the Arabidopsis thaliana ribulose-1,5-bisphosphate carboxylase small subunit promoter (Rubisco SSU)* and terminates at the Nicotiana tabacum ribulose-1,5-bisphosphate carboxylase small subunit terminator. The nucleotide sequence of cry1Ab, originating from Bacillus thuringiensis was codon optimized for plant expression and encodes a 615-amino acid protein (68.9 kDa), corresponding to the trypsin-resistant, insecticidally active core protein, following cleavage of the 1155-amino acid native Cry1Ab protoxin.

For selection during cloning, a kanamycin resistance gene cassette was also integrated into the genome. Transcription of the neomycin phosphotranferase II (nphII) gene starts at the subterranean clover stunt virus (SCSV) 1 promoter. Two segments of nptII are transcribed, with a catalase-1 intron from Ricinus communis between the two segments and an intervening sequence from the MB4 plasmid following the second segment. Transcription stops at the SCSV3 terminator.

DNA insertion from pMB6:
Transcription of the cry2Ab gene occurs from the A. thaliana Rubisco SSU promoter* and terminates at the N. tabacum Rubisco SSU terminator. A chloroplast targeting peptide sequence has been added before the cry2Ab sequence to localize the translated protein to the chloroplasts. The protein coding region of cry2Ab is derived from B. thuringiensis ssp. kurstaki HD-1. The bacterial gene sequence was optimized for efficient expression in plants by the removal of polyadenylation signals, lowering of the AT content, and the removal of destabilising sequences.

A second nphII kanamycin resistance cassette was also integrated into the genome. Transcription also commences from the SCSV1 promoter** and terminates at the SCSV3 terminator, which contains a single polyadenylation signal.

Note:
* The Rubisco SSU promoter contains 1.7 kb of 5' untranslated region of the gene.
** No enhancers present
LMO characteristics
Modified traits
Common use(s)
  • Food