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


Living Modified Organism (LMO)

Decisions on the LMO Risk Assessments  
last updated: 12 Feb 2014
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
Event 15985 (tradename Bollgard II®) was derived through the retransformation of transgenic cotton line MON 531 (AKA DP50B), by biolistic transformation with plasmid DNA containing the cry2Ab gene originally isolated from Bacillus thuringiensis subsp. kurstaki.

As a result, event 15985 expresses both the Cry1Ac and Cry2Ab insecticidal proteins. Cotton line 15985 is intended to protect cotton from feeding by a range of Lepidopteran species 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).

Monsanto has proposed cotton line 15985, with its two insecticidal proteins, as a means to provide more effective insect resistance management.
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.
Variety: DP50B
Characteristics of the modification process
PV-GHBK11 (PV-GHBK04 pre-existing in parental organism)
  • 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)
  • BCH-GENE-SCBD-103901-2 HSP 70 5' untranslated leader sequence | Petunia hybrida (Petunia, PETHY)
  • 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)
  • BCH-GENE-SCBD-103856-6 α' subunit of β-conglycinin gene terminator | Glycine max (Soybean, Soya bean, Soya, SOYBN)
  • 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)
Information on the inserted DNA sequences from vector PV-GHBK11
In addition to the Cry1A(c) coding region that is also present in the Bollgard™ cotton (vector PV-GHBK04), this LMO also contains an HPLC-isolated linear restriction fragment of the plasmid vector, designated PV-GHBK11, utilized for transformation of Bollgard II cotton event 15985, contains one copy of both the cry2Ab2 and uidA plant gene expression cassettes.

The expression cassette that produces the Cry2Ab2 protein consists of the enhanced 35S promoter, the cry2Ab coding sequence that was modified for optimal expression in plants, and the 3’ non translated region of the nopaline synthase (nos) gene from Agrobacterium tumefaciens which provides the signal for mRNA polyadenylation.

Southern blot analysis indicated that a single T-DNA construct had integrated into the 15985 line. The 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 vector PV-GHBK04
The parental organism (MON531) was shown to have integrated one complete and one partial copy of the T-DNA expression cassette. This cassette contains and expresses the cry1A(c)  gene and the nptII resistance gene. The aadA gene was also integrated into the host genome however it is not expressed.

For additional information on this LMO, please refer to the records of the parental LMO.
LMO characteristics
  • Food
  • Feed
  • Fiber/textile
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, line 15985 also contains the nptII and aad selectable marker genes (derived from the parental cotton line containing event 531) and the beta-D-glucuronidase (GUS) encoding uidA gene from Escherichia 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.