MON-15985-7 - Bollgard II™ cotton | BCH-LMO-SCBD-14774 | Living Modified Organism | Biosafety Clearing-House

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Living Modified Organism (LMO)

Decisions on the LMO Risk Assessments  
published: 05 Jun 2006 last updated: 23 Jan 2023
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.
Bollgard II™ cotton
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MON15985
Yes
MON-15985-7
Event MON15985 (tradename Bollgard II®) was derived through the transformation of modified cotton line MON531. The modified cotton expresses Bacillus thuringiensis subsp. kurstaki crystal proteins Cry1Ac and Cry2Ab2, which confer resistance to Lepidopteran pests, including tobacco budworm (Heliothis virescens), pink bollworm (Pectinophora gossypiella), cotton bollworm (Helicoverpa zea), cabbage looper (Trichoplusia ni), saltmarsh caterpillar (Estigmene acrea), cotton leaf perforator (Bucculatrix thurbeiella), soybean looper (Pseudoplusia includens), beet armyworm (Spodoptera exigua), fall armyworm (Spodoptera frugiperda), yellowstriped armyworm (Spodoptera ornithogolli) and European corn borer (Ostrinia nubilalis), among others. In addition to the insecticidal proteins, the modified cotton also contains two selectable markers from Escherichia coli: neomycin phosphotransferase II (antibiotic selection during transformation of the parental plant) and beta-glucuronidase (colorimetric selection during second transformation).
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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-14775-16 Living Modified Organism MON-ØØ531-6 - Bollgard™ cotton
    Monsanto | Resistance to antibiotics (Kanamycin, Streptomycin), Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
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Characteristics of the modification process
PV-GHBK11; PV-GHBK04
EN
  • Biolistic / Particle gun
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
  • BCH-GENE-SCBD-14988-7 Cry2Ab2 | Bacillus thuringiensis (Bt, Bacillus, BACTU)
    Protein coding sequence | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • BCH-GENE-SCBD-15033-8 3"(9)-O-aminoglycoside adenyltransferase | Escherichia coli (ECOLX)
    Protein coding sequence | Resistance to antibiotics (Streptomycin)
  • BCH-GENE-SCBD-14986-6 Cry1Ac | Bacillus thuringiensis (Bt, Bacillus, BACTU)
    Protein coding sequence | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • BCH-GENE-SCBD-46004-7 Beta-glucuronidase coding sequence | Escherichia coli (ECOLX)
    Protein coding sequence | Selectable marker genes and reporter genes
  • BCH-GENE-SCBD-100366-6 CaMV Enhanced 35S promoter | Cauliflower mosaic virus (CaMV)
    Promoter
  • BCH-GENE-SCBD-103901-2 HSP 70 5' untranslated leader sequence | Petunia hybrida (Petunia, PETHY)
    Leader
  • BCH-GENE-SCBD-100365-6 Chloroplast transit peptide 2 | Arabidopsis thaliana (Thale cress, Mouse-ear cress, Arabidopsis, ARATH)
    Transit signal
  • BCH-GENE-SCBD-100269-8 Nopaline Synthase Gene Terminator | Agrobacterium tumefaciens (Agrobacterium)
    Terminator
  • BCH-GENE-SCBD-103856-6 α' subunit of β-conglycinin gene terminator | Glycine max (Soybean, Soya bean, Soya, SOYBN)
    Terminator
  • BCH-GENE-SCBD-15001-5 Neomycin Phosphotransferase II | Escherichia coli (ECOLX)
    Protein coding sequence | Resistance to antibiotics (Kanamycin)
  • BCH-GENE-SCBD-100287-7 CaMV 35S promoter | Cauliflower mosaic virus (CaMV)
    Promoter
  • BCH-GENE-SCBD-101416-6 Ti plasmid right border repeat | Agrobacterium tumefaciens (Agrobacterium)
    Plasmid vector
Information on the inserted DNA sequences from vector PV-GHBK11
Transformation with vector PV-GHBK11 inserted two gene cassettes into the modified parental line MON531: Bacillus thuringiensis cry2Ab2 and Escherichia coli beta-glucuronidase (uidA).

The cry2Ab2 sequence is under control of a Cauliflower mosaic virus (CaMV) 35S enhanced promoter and an Agrobacterium tumefaciens nopaline synthase (nos) terminator. A Petunia hybrida heat shock protein 70 leader for enhanced expression and an Arabidopsis thaliana chloroplast transit peptide 2 for targeting translated protein to the chloroplast are also present.

The uidA coding sequence is under control of the CaMV 35S enhanced promoter and nos terminator.

Note:
  • Due to the constitutive nature of the viral promoters, high levels of expression are expected in all plant tissues.
  • Southern blot analysis indicated that a single T-DNA construct had integrated into the genome. The CaMV 35S promoter that drives the expression of the uidA gene was truncated, however this did not affect the expression of the gene. The analysis also showed that there was no integration of any portions of the vector backbone.

Pre-existing DNA insert from MON531 vector PV-GHBK04
The parental organism (MON531) contained two gene cassettes: B. thuringiensis cry1Ac and E. coli neomycin phosphotransferase II (nptII).

The cry1Ac coding sequence is under control of CaMV 35S enhanced promoter and Glycine max α' subunit of β-conglycinin terminator. See notes below about the partial cry1Ac cassette and antibiotic resistance gene aad.

The nptII coding sequence is under control of a CaMV 35S promoter and and Agrobacterium tumefaciens nopaline synthase terminator.

Note:
  • Due to the constitutive nature of the viral promoters, high levels of expression are expected in all plant tissues.
  • The cry1Ac coding sequence was codon-optimized for expression in plants. The codon optimization resulted in a single amino acid change (leucine substituted for serine) at position 766 (L766S). The sequence was originally sourced from B. thuringiensis subsp. kurstaki HD-73.
  • Southern blot analysis indicated that the LMO contains a single active copy and a partial inactive copy of the cry1Ac gene both of which are linked.
  • The T-DNA contains the antibiotic resistance 3"(9)-O-aminoglycoside adenylyltransferase (aad) gene, which facilitated the selection of bacteria containing the plasmid in the initial steps of transforming the cotton tissue. The gene is under control of a bacterial promoter, which is not active in plant tissues.  
  • For additional information, please also refer to the parental MON531 record.
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LMO characteristics
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  • Feed
  • Fiber/textile
  • Food
Detection method(s)
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Additional Information
As with other B. thuringiensis-derived delta-endotoxins, the Cry1Ac and Cry2Ab proteins exert their insecticidal activity by binding to specific receptors located on the brush border midgut epithelium of susceptible insect species. Following binding, cation-specific pores are formed that disrupt midgut ionic equilibrium leading to gut paralysis and eventual death due to bacterial sepsis. Cry1Ac and Cry2Ab are highly selective and are only active against Lepidopteran insects. These proteins do, however, interact with different receptor sites in the target insects and it is expected that “stacking” these traits will result in increased protection against insect attack and a delay in the development of resistant insect populations.

In addition to the cry genes conferring insect resistance, MON15985 also contains the nptII and aad selectable marker genes (derived from the parental cotton line containing event MON531) and the beta-D-glucuronidase (GUS) encoding uidA gene from E. coli. This latter gene was introduced as a visually scorable marker gene to identify transformed plantlets in tissue culture. The GUS enzyme can be used to catalyze a colorimetric reaction resulting in the production of a blue colour in transformed cells.
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