BCS-BNØ12-7 × ACS-BNØØ3-6 × MON-883Ø2-9 - Fertility restored, Herbicide tolerant canola | BCH-LMO-SCBD-115686 | Living Modified Organism | Biosafety Clearing-House

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Living Modified Organism (LMO)
  |  
Decisions on the LMO Risk Assessments  
last updated: 25 Aug 2020
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.
Fertility restored, Herbicide tolerant canola
EN
MS11 × RF3 × MON88302
Yes
BCS-BNØ12-7 × ACS-BNØØ3-6 × MON-883Ø2-9
The modified canola (Brassica napus) was created through crossing of three modified canola lines for tolerance to herbicides. For tolerance to glufinosate, the modified canola expresses Streptomyces hygroscopicus phosphinothricin N-acetyltransferase, which catalyzes the acetylation of phosphinothricin to prevent glutamine synthetase inhibition. For tolerance to glyphosate, the modified canola expresses Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase, which does not bind glyphosate and allows for the continued biosynthesis of essential aromatic amino acids via the shikimate pathway. The modified canola additionally contains the engineered restoration of male fertility from a parental hybrid line. The canola expresses Bacillus amyloliquefaciens barnase, an RNase, in the tapetum cells of the pollen sac during anther development and causes male sterility by interfering with RNA production. However, the canola also expresses B. amyloliquefaciens barstar, which inhibits barnase by forming a stable complex with the enzyme. Thus, barnase activity is prevented in pollen tissues and male fertility is restored.
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-12083-7 Organism Brassica napus (Turnip, Rapeseed, Canola Plant, Oilseed Rape, Rape, BRANA)
    Crops
  • BCH-LMO-SCBD-112988-2 Living Modified Organism BCS-BNØ12-7 - Male Sterile/ Fertility Restored Herbicide Tolerant Canola
    Bayer CropScience | Changes in physiology and/or production (Reproduction, Male sterility), Resistance to herbicides (Glufosinate)
  • BCH-LMO-SCBD-14755-7 Living Modified Organism ACS-BNØØ3-6 - InVigor™ canola This document has been updated. This is not the latest published version. Click here to view the latest version of the record.
    Bayer CropScience | Changes in physiology and/or production (Fertility restoration), Resistance to herbicides (Glufosinate)
  • BCH-LMO-SCBD-104752-4 Living Modified Organism MON-883Ø2-9 - TruFlex Roundup Ready™ Canola
    Monsanto | Resistance to herbicides (Glyphosate)
Please note the developer has not been confirmed.
EN
Characteristics of the modification process
pTCO113; pTHW118; PV-BNHT2672
EN
  • Cross breeding
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
  • BCH-GENE-SCBD-103067-9 Transcript 7 gene 3' untranslated region | Agrobacterium tumefaciens (Agrobacterium)
    Terminator
  • BCH-GENE-SCBD-14972-12 Phosphinothricin N-acetyltransferase gene | Streptomyces hygroscopicus (STRHY)
    Protein coding sequence | Resistance to herbicides (Glufosinate)
  • BCH-GENE-SCBD-103851-5 rbcS Promoter | Arabidopsis thaliana (Thale cress, Mouse-ear cress, Arabidopsis, ARATH)
    Promoter
  • BCH-GENE-SCBD-100269-8 Nopaline Synthase Gene Terminator | Agrobacterium tumefaciens (Agrobacterium)
    Terminator
  • BCH-GENE-SCBD-104825-2 Barnase 3' Untranslated region | Bacillus amyloliquefaciens (BACAM)
    Terminator
  • BCH-GENE-SCBD-14973-6 Barnase | Bacillus amyloliquefaciens (BACAM)
    Protein coding sequence | Changes in physiology and/or production (Reproduction, Male sterility)
  • BCH-GENE-SCBD-101407-6 pTA29 pollen specific promoter | Nicotiana tabacum (Tobacco, TOBAC )
    Promoter
  • BCH-GENE-SCBD-100270-6 Nopaline Synthase Gene Promoter | Agrobacterium tumefaciens (Agrobacterium)
    Promoter
  • BCH-GENE-SCBD-14974-7 Barstar | Bacillus amyloliquefaciens (BACAM)
    Protein coding sequence | Changes in physiology and/or production (Fertility restoration)
  • BCH-GENE-SCBD-101409-2 Barstar gene terminator | Bacillus amyloliquefaciens (BACAM)
    Terminator
  • BCH-GENE-SCBD-105196-2 FMV 35S Enhancer | Figwort mosaic virus (Figwort mottle virus, FMV, CMoVb)
    Leader
  • BCH-GENE-SCBD-103903-1 Elongation factor EF-1alpha promoter | Arabidopsis thaliana (Thale cress, Mouse-ear cress, Arabidopsis, ARATH)
    Promoter
  • BCH-GENE-SCBD-103904-1 Elongation factor EF-1alpha Leader | Arabidopsis thaliana (Thale cress, Mouse-ear cress, Arabidopsis, ARATH)
    Leader
  • BCH-GENE-SCBD-103905-1 Elongation factor EF-1alpha Intron 1 | Arabidopsis thaliana (Thale cress, Mouse-ear cress, Arabidopsis, ARATH)
    Intron
  • BCH-GENE-SCBD-100365-6 Chloroplast transit peptide 2 | Arabidopsis thaliana (Thale cress, Mouse-ear cress, Arabidopsis, ARATH)
    Transit signal
  • BCH-GENE-SCBD-14979-7 5-enolpyruvylshikimate-3-phosphate synthase gene | Agrobacterium tumefaciens (Agrobacterium)
    Protein coding sequence | Resistance to herbicides (Glyphosate)
  • BCH-GENE-SCBD-101877-5 rbcS-E9 gene terminator | Pisum sativum (Garden pea, PEA)
    Terminator
DNA insert from MS11 vector pTCO113
The DNA insert from pTCO113 contains three gene cassettes: Streptomyces hygroscopicus phosphinothricin N-acetyltransferase (bar); Bacillus amyloliquefaciens barnase and B. amyloliquefaciens barstar.

Transcription of bar commences from the Arabidopsis thaliana ribulose-1,5-bisphosphate carboxylase small subunit (rbcS) promoter  and terminates at the Agrobacterium tumefaciens transcript 7 3' untranslated region.

Transcription of barnase starts from the Nicotiana tabacum pTA29 promoter and ends at the A. tumefaciens nopaline synthase (nos) terminator. The transcript additionally contains the 3' untranslated region of Barnase. The pTA29 promoter is pollen specific and transcription is expected to occur only within pollen tissues.

Transcription of barstar commences from the nos promoter and terminates at a transcript 7 3' untranslated region.

Note:
- DNA sequencing and southern blot analyses indicated that the parental genome contain a single, intact T-DNA insert

DNA insert from RF3 vector pTHW118
The DNA insert from the pTHW118  contains two gene cassettes: barstar and bar.

Transcription of barstar starts from the pTA29 promoter and ends at a nos terminator. The transcript additionally contains the barstar gene terminator, which is not expected to be translated into protein.The pTA29 promoter is pollen specific and transcription is expected to occur only within pollen tissues.

Transcription of bar commences from the rbcS promoter and terminates at the transcript 7 3' untranslated region.

Note:
- Two codons on the N-terminus of bar have been substituted for the codons ATG and GAC.
- Southern blot and PCR anaylses indicated that the parental genome contains one complete copy of the transformation cassette and an inverted partial copy containing a portion of the pTA29 promoter, barstar and the nos terminator adjacent to the left T-DNA boundary.

DNA insert from MON88302 vector PV-BNHT2672
The DNA insert from PV-BNHT2672 contains a single gene cassette for expression of Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (epsps). Transcription of epsps is under control of a chimeric promoter, containing the Figwort mosaic virus 35S enhancer and the Arabidopsis thaliana elongation factor 1-alpha (EF-1α), and Pisum sativum ribulose-1,5-bisphosphate carboxylase small subunit E9 terminator (rbcS-E9). The transcript initially contains an A. thaliana EF-1α leader, an EF-1α intron 1 and an A. thaliana chloroplast transit peptide 2 prior (5') to the epsps coding sequence. High levels of transcription are expected from the chimeric promoter. The EF-1α leader and intron additionally enhance expression. The chloroplast transit peptide targets EPSPS for accumulation within the chloroplast.

Note:
- Southern blot analysis indicated that the parental genome contains a single T-DNA insertion without backbone integration.

For more information, kindly refer to the parental LMO records.
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LMO characteristics
EN
  • Food
  • Feed
Additional Information
Barnase ribonuclease is secreted by B. amyloliquefaciens and is lethal due its RNase activity. In the bacterium, the inhibitor, barstar, is also synthesized and binds to barnase after synthesis to prevent damage due to degradation of the cellular RNA. The inhibitor is removed upon secretion.

Glyphosate binds the endogenous 5-enolpyruvulshikimate-3-phosphate synthase enzyme, inactivating the enzyme and preventing the essential biosynthesis of aromatic amino acids (tyrosine, phenylalanine and tryptophan) and folates via the shikimate pathway. The introduction of a bacterial epsps coding sequence allows for the production of aromatic compounds in the presence of glyphosate as the bacterial EPSPS protein will not bind the compound.
EN
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