DAS-24236-5 × DAS-21Ø23-5 × SYN-IR1Ø2-7 × MON-88913-8 × DAS-8191Ø-7 - Insect resistant, herbicide tolerant cotton | BCH-LMO-SCBD-109709 | Living Modified Organism | Biosafety Clearing-House

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Living Modified Organism (LMO)

Decisions on the LMO Risk Assessments  
last updated: 17 Jun 2022
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.
Insect resistant, herbicide tolerant cotton
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281-24-236 × 3006-210-23 × MON88913 × COT102 × DAS81910
DAS-24236-5 × DAS-21Ø23-5 × SYN-IR1Ø2-7 × MON-88913-8 × DAS-8191Ø-7
The stacked cotton (Gossypium hirsutum) line was produced by cross breeding between each of the parental organisms for insect resistance and herbicide tolerance.  For Lepidopteran resistance, the modified cotton expresses Bacillus thuringiensis Cry1Ac, Cry1F and Vegetative insecticidal protein 3A. For tolerance to glufosinate, the modified cotton expresses Streptomyces viridochromogenes phosphinothricin N-acetyltransferase and Streptomyces hygroscopicus phosphinothricin N-acetyltransferase. For tolerance to 2,4-dichlorophenoxyacetic acid, the cotton expresses Delftia acidovorans aryloxyalkanoate dioxygenase. For tolerance to glyphosate, the cotton expresses Rhizobium radiobacter 5-enolpyruvylshikimate-3-phosphate synthase.  Additionally, an Escherichia coli hygromycin B phosphotransferase cassette is also present for hygromycin selection during transformation and breeding.
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.
  • BCH-ORGA-SCBD-12080-6 Organism Gossypium hirsutum (Cotton)
    Crops
  • BCH-LMO-SCBD-14938-7 Living Modified Organism DAS-21Ø23-5 - Insect-resistant cotton
    Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • BCH-LMO-SCBD-14940-7 Living Modified Organism DAS-24236-5 - Insect-resistant cotton
    Dow AgroSciences | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • BCH-LMO-SCBD-15168-16 Living Modified Organism MON-88913-8 - Roundup Ready™ Flex™ cotton
    Resistance to herbicides - Glyphosate
  • BCH-LMO-SCBD-14992-8 Living Modified Organism 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
  • BCH-LMO-SCBD-108872-2 Living Modified Organism DAS-8191Ø-7 - Cotton modified for herbicide tolerance
    XXXX | Resistance to herbicides (Glufosinate), Tolerance to 2,4-Dichlorophenoxyacetic acid
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Characteristics of the modification process
pAGM281; pMYC3006; pCOT-1; PV-GHGT35 and pDAB4468
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  • Cross breeding
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
DNA insert from DAS-24236-5 vector pAGM281
The DNA insert from 281-24-236 contains two gene cassettes: Bacillus thuringiensis var. aizawai cry1F and  Streptomyces viridochromogenes phosphinothricin N-acetyltransferase (pat).

Transcription of pat is under control of a Zea mays ubiquitin promoter and an Agrobacterium tumefaciens open reading frame 25 (ORF25) terminator. The transcript is expected to initially contain a Z. mays ubiquitin intron 1 at the 5’ end. The intron enhances the expression of pat.

Transcription of cry1F is under control of a synthetic 4ocs∆Mas2' promoter and the same ORF25 terminator as the pat expression cassette. The cassette is in the reverse orientation to utilize the same terminator.

Note:
  • Transcription is expected to occur at elevated and constitutive levels due to the promoters.
  • The coding sequences of pat and cry1F have been optimized for expression in plants
  • Cry1F is a chimeric, full-length δ-endotoxin comprised of the core toxin of Cry1F and nontoxic portions of B. thuringiensis Cry1Ca3 and Cry1Ab1 proteins, which form the C-terminal end of the protein and are removed during the activation of the Cry1F protein.

DNA insert from  DAS-21Ø23-5  vector pMYC3006
The DNA insert from 3006-210-23 contains two gene cassettes: Bacillus thuringiensis cry1Ac and Streptomyces viridochromogenes phosphinothricin N-acetyltransferase (pat).

Transcription of pat is under control of a synthetic 4ocs∆Mas2' promoter and an Agrobacterium tumefaciens open reading frame 25 (ORF25) terminator. Transcription of cry1Ac is under control of a Zea mays ubiquitin promoter and the same ORF25 terminator as the pat expression cassette.

Note:
  • The ORF25 terminator serves as the terminator for both gene expression cassettes. The cry1Ac cassette is in the reverse orientation to utilize the same terminator.
  • The coding sequences of cry1Ac and pat have been optimized for expression in plant cells.
  • Transcription is expected to occur at elevated and constitutive levels due to the promoters.
  • Southern blot analysis demonstrated that a single, intact DNA insertion was present in the parental genome. The analysis also indicated that no vector backbone sequences were integrated during transformation.

DNA insert from SYN-IR1Ø2-7 vector pCOT-1
The DNA insert from COT102 contains two gene cassettes: Escherichia coli hygromycin B phosphotransferase (hph) and Bacillus thuringiensis vegetative insecticidal protein 3A.

Transcription of hph is under control of the Arabidopsis thaliana ubiquitin 3 promoter and the Agrobacterium tumefaciens nopaline synthase (nos) terminator. The gene cassette is present in the counterclockwise orientation.

The expression of Bacillus thuringiensis vegetative insecticidal protein 3A (vip3A) is under transcriptional control of the A. thaliana actin 2 promoter and the nos terminator.

Note:
  • The coding sequence of the vip3A was altered for optimal expression in plant cells.
  • Transcription is expected to occur at elevated and constitutive levels due to the promoters.
  • Southern blot analysis indicated the incorporation of a single copy of the transgenes without the integration of the vector backbone sequences.
  • Southern blot analysis confirmed the expression of the proteins.

DNA insert from MON-88913-8 vector PV-GHGT35
The DNA insert from MON88913 contains two codon-optimised Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (epsps) cassettes:
  1. the first epsps coding sequence under the regulation of a chimeric transcriptional promoter (Figwort mosaic virus 34S promoter enhancer and Arabidopsis thaliana elongation factor EF-1 alpha (tsf1) promoter), tsf1 leader and intron sequences, an A. thaliana chloroplast transit peptide 2 sequence and a Pisum sativum ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) E9 transcript termination and polyadenylation sequence (T-E9).
  2. the second epsps coding sequence regulated by another chimeric transcriptional promoter (Cauliflower mosaic virus 35S enhancer and A. thaliana actin 8 (act8) promoter), act8 leader and intron sequences, A. thaliana chloroplast targeting peptide 2 and T-E9.

High levels of transcription are expected from both cassettes due to the presence of viral enhancer sequences. The EPSPS protein is expected to accumulate in the chloroplast due to the transit signal peptide.

Note:
The promoter for both genetic constructs are chimeric promoters containing viral enhancer sequences. Thus, the size of the promoters (Elongation factor 1 alpha and Actin 8) in the 'Genetic elements construct' reflects the size of chimeric promoters (FMV 35S enhancer + Elongation factor 1 alpha promoter; CaMV 35S enhancer + Actin 8 promoter).

DNA insert from  DAS-8191Ø-7 vector DAB4468
The DNA insert from DAS81910 contains two gene cassettes: Delftia acidovorans aryloxyalkanoate dioxygenase (aad-12) and Streptomyces hygroscopicus phosphinothricin N-acetyltransferase (pat).

Transcription of aad-12 is under control of the Arabidopsis thaliana polyubiquitin 10 promoter and the Agrobacterium tumefaciens open reading frame 23 3’ untranslated region. Transcription of pat is under control of the Cassava Vein Mosaic Virus promoter and the A. tumefaciens ORF1 3’ untranslated region.

Note:
  • Transcription is expected to occur at elevated and constitutive levels due to the promoters.
  • Southern blot analysis indicated a single, intact insertion of aad-12 and pat into the parental genome without the integration of the vector backbone sequences.

For more information, kindly refer to the parental LMO records.
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LMO characteristics
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  • Fiber/textile