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Modified Organism
BCS-GHØØ4-7 x BCS-GHØØ5-8 x SYN-IR1Ø2-7 - Insect-resistant, herbicide-tolerant cotton
Record information and status
Record ID
115044
Status
Published
Date of creation
2019-07-10 19:30 UTC (austein.mcloughlin@cbd.int)
Date of publication
2019-07-10 19:30 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
Insect-resistant, herbicide-tolerant cotton
Transformation event
T304-40 x GHB 119 x COT102
Unique identifier
BCS-GHØØ4-7 x BCS-GHØØ5-8 x SYN-IR1Ø2-7
Developer(s)
BASF Plant Science GmbH
Carl-Bosch-Str. 38
Ludwigshafen
Germany, 67056
Phone:+49 621 60-0
Fax:+49 621 60-42525
Email:global.info@basf.com
Url:BASF AG - Pflanzenbiotechnologie
Description
The modified cotton was produced cross-breeding to obtain a stacked event with insect resistance and herbicide tolerance. The plant expresses the Bacillus thuringiensis Cry1Ab (Pesticidal crystal protein Cry1Ab), Cry2Ae and VIP3A proteins, which confers resistance to Lepidopteran insect larvae, such as cotton bollworm larvae (Helicoverpa zea), tobacco budworm larvae (Heliothis virescens) and fall armyworm larvae (Spodoptera frugiperda). In particular, VIP3A was included to prevent and manage resistance developing against the crystal proteins. Additionally, the modified cotton expresses the proteins phosphinothricin acetyltransferase from Streptomyces hygroscopicus, which confers tolerance to herbicides containing glufosinate ammonium. The selectable marker hygromycin B phosphotransferase for hygromycin resistance is also present.
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.
BCS-GHØØ4-7 - Herbicide-tolerant, insect-resistant cotton
Resistance to diseases and pests - Insects - Lepidoptera (butterflies and moths) Resistance to herbicides - Glufosinate
Show detection method(s)
BCS-GHØØ5-8 - Herbicide-tolerant and lepidoptera-resistant cotton
Bayer BioScience N.V. Resistance to diseases and pests - Insects - Lepidoptera (butterflies and moths) - Cotton bollworm (Helicoverpa spp.), armyworm (Spodoptera frugiperda) Resistance to herbicides - Glufosinate
Show detection method(s)
SYN-IR1Ø2-7 - VIPCOT™ Cotton
Resistance to antibiotics - Hygromycin Resistance to diseases and pests - Insects - Lepidoptera (butterflies and moths) Selectable marker genes and reporter genes
Show detection method(s)
Characteristics of the transformation process
Vector
pTDL008; pTEM12; pCOT-1
Techniques used for the modification
  • Cross breeding
Genetic elements construct
 
NADP-malic enzyme 1 gene 3’UTR and terminator
0.94 Kb
 
 
Cry1Ab
1.85 Kb
 
 
5'e1 Leader
0.06 Kb
 
 
Ps7s7
1.04 Kb
 
 
CaMV 35S promoter
0.86 Kb
 
 
Phosphinothricin N-acetyltransferase gene
0.55 Kb
 
 
Nopaline Synthase Gene Terminator
0.31 Kb
 
 
Ti plasmid left border repeat
0.02 Kb
 
 
Nopaline Synthase Gene Terminator
0.31 Kb
 
 
Phosphinothricin N-acetyltransferase gene
0.55 Kb
 
 
CsVMV promoter
0.54 Kb
 
 
CaMV 35S promoter
0.48 Kb
 
 
5' untranslated leader of chlorophyll a/b-binding protein
0.07 Kb
 
 
rbcS Transit Peptide
0.16 Kb
 
 
Cry2Ae
1.90 Kb
 
 
CaMV 35S terminator
0.27 Kb
 
 
Ti plasmid right border repeat
0.02 Kb
 
 
Nopaline Synthase Gene Terminator
0.25 Kb
 
 
Hygromycin B phosphotransferase gene
1.03 Kb
 
 
Ubiquitin gene 3 promoter
1.72 Kb
 
 
Actin 2 Gene Promoter
1.41 Kb
 
 
Vegetative insecticidal protein 3A
2.37 Kb
 
 
Nopaline Synthase Gene Terminator
0.25 Kb
 
Further details
Notes regarding the genetic elements introduced or modified in this LMO
Genetic elements introduced from pTDL008 (derived from pGSV20):
Transcription of the Bacillus thuringiensis Cry1Ab gene begins at the Subterranean clover stunt virus promoter (Ps7s7) and terminates at the Flavera bidentis NADP-malic enzyme 1 (me1) terminator. A 5' leader sequence from Oryza sativa GE1 was included to enhance transcription. The gene cassette is in the counter-clockwise orientation.

A second gene cassette contains a herbicide tolerance gene, Streptomyces hygroscopicus phosphinothricin N-acetyltransferase gene (bar), which is under transcriptional control of the Cauliflower mosaic virus (CMV) 35S promoter and the Agrobacterium tumefaciens nopaline synthase (nos) terminator.

Note:
Sequencing of the 9056 bp inserted transgenic construct and Southern blot analysis revealed an almost full copy of the T-DNA construct (with an incomplete 3`me1 terminator) was inserted into the T304-40 LM cotton line in addition to:
- a partial 3' me1 terminator;
- a partial copy of the cry1Ab gene cassette, with a truncated Ps7s7 promoter, in a tail-to-tail orientation, and
- a partial copy of the bar gene cassette in which the nos terminator is truncated.

pTEM12 (derived from pGSC1700):
Another bar gene cassette was integrated into the parental line in a counter-clockwise orientation. Transcription commences from the Cassava vein mosaic virus promoter and terminates the CMV 35S terminator.

A Cry2Ae gene cassette was also integrated into the parental line. Transcription of the B. thuringiensis Cry2Ae gene is controlled by the CaMV 35S promoter and terminator. In addition to the Cry2Ae coding sequence, the transcript contains the 5' untranslated leader of Petunia hybrida chlorophyll a/b-binding protein and the Arabidopsis thaliana ribulose-1,5-bisphosphate carboxylase small subunit transit peptide for enhancing transcription and higher steady-state levels of the protein in the leaves, respectively.

Note:
Southern Blot analysis indicated that a single copy of the insert is present in the GHB119 line and a configuration that correspond to that in the original vector. No insertion of plasmid backbone sequences was detected.

pCOT-1:
Transcription of the Escherichia coli hygromycin B phosphotransferase is under control of the A. thaliana ubiquitin 3 promoter and the nos terminator. The gene cassette is present in the counterclockwise orientation. The hygromycin B phosphotransferase (aph4) gene is used for selection of transformants.

The expression of B. thuringiensis vegetative insecticidal protein 3A (vip3A) is under control of the A. thaliana actin 2 promoter and the nos terminator. Coding sequence of the Vip3A gene was altered for optimal expression in plant cells. Southern blot analysis confirmed the protein expression.

Note:
Southern blot analysis of the genomic DNA from COT102 revealed the incorporation of single intact copies of the vip3Aa and aph4, along with intact copies of their respective regulatory sequences. Results of these analyses also demonstrated that none of the vector backbone sequences, including the streptomycin adenyltransferase gene, were incorporated into the genomic DNA.

For more information, kindly review the records of the parental lines.
LMO characteristics
Modified traits
Common use(s)
  • Food
  • Feed
  • Fiber / Textile