ACS-BNØØ3-6 × MON-ØØØ73-7 - Herbicide tolerant, male fertility restoring canola | BCH-LMO-SCBD-116285 | Living Modified Organism | Biosafety Clearing-House

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Living Modified Organism (LMO)
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Decisions on the LMO Risk Assessments  
last updated: 27 Sep 2021
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Herbicide tolerant, male fertility restoring canola
EN
RF3 × RT73
Yes
ACS-BNØØ3-6 × MON-ØØØ73-7
The modified canola (Brassica napus) was produced through the cross-breeding of modified parental lines for male fertility restoration and herbicide tolerance. The canola expresses Bacillus amyloliquefaciens barstar, an inhibitor of the ribonuclease barnase. In lines where barnase is expressed, barstar prevents the RNA degradation in the pollen tissues and thus restores male fertility (pollen development). For herbicide tolerance, the canola expresses Streptomyces hygroscopicus phosphinothricin acetyltransferase (glufosinate tolerance), Agrobacterium tumefaciens 5-enolpyruvylshikimate- 3-phosphate synthase (glyphosate tolerance) and Ochrobactrum anthropi glyphosate oxidoreductase (glyphosate tolerance).
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.
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Characteristics of the modification process
pTHW118; PV-BNGT04
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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.
  • BCH-GENE-SCBD-101407-6 pTA29 pollen specific promoter | Nicotiana tabacum (Tobacco, TOBAC )
    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-100269-8 Nopaline Synthase Gene Terminator | Agrobacterium tumefaciens (Agrobacterium)
    Terminator
  • BCH-GENE-SCBD-103851-5 rbcS Promoter | Arabidopsis thaliana (Thale cress, Mouse-ear cress, Arabidopsis, ARATH)
    Promoter
  • BCH-GENE-SCBD-14972-12 Phosphinothricin N-acetyltransferase gene | Streptomyces hygroscopicus (STRHY)
    Protein coding sequence | Resistance to herbicides (Glufosinate)
  • BCH-GENE-SCBD-103067-9 Transcript 7 gene 3' untranslated region | Agrobacterium tumefaciens (Agrobacterium)
    Terminator
  • BCH-GENE-SCBD-101507-5 FMV 34S promoter | Figwort mosaic virus (Figwort mottle virus, FMV, CMoVb)
    Promoter
  • 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
  • BCH-GENE-SCBD-101902-4 rbcS Transit Peptide | Arabidopsis thaliana (Thale cress, Mouse-ear cress, Arabidopsis, ARATH)
    Transit signal
  • BCH-GENE-SCBD-14998-4 Glyphosate oxidoreductase gene | Ochrobactrum anthropi (OCHAN)
    Protein coding sequence | Resistance to herbicides (Glyphosate)
DNA insert from pTHW118 from RF3 (ACS-BNØØ3-6) canola
The DNA insert contains two gene cassettes: Bacillus amyloliquefaciens barstar and Streptomyces hygroscopicus phosphinothricin N-acetyltransferase (bar).

Barstar is under control of a Nicotiana tabacum TA29 pollen specific promoter and an Agrobacterium tumefaciens nopaline synthase terminator. An additional sequence, B. amyloliquefaciens barstar 3’ untranslated region, which contributes to the polyadenylation of the coding sequence, can be found between the barstar coding sequence and nopaline synthase terminator.

Phosphinothricin N-acetyltransferase is under control of an Arabidopsis thaliana ribulose-1,5-bisphosphate carboxylase (Rubisco) small subunit promoter and A. tumefaciens transcript 7 3’ untranslated region.

Note:
- The coding sequence of phosphinothricin N-acetyltransferase has the two N-terminal codons modified to ATG and GAC.
- Southern blot and PCR analysis indicated that the parental genome contains one complete copy of the transformation cassette arranged in an inverted repeat orientation to a second partial copy (comprising of a portion of the TA29 promoter, the barstar coding sequence, the nos terminator sequence and a non-initiation codon of the bar gene)  of the transformation cassette adjacent to the left T-DNA boundary.


DNA insert from PV-BNGT04 from RT73 (MON-ØØØ73-7) canola
The DNA insert contains two gene cassettes: Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps) and Ochrobactrum anthropic glyphosate oxidoreductase (gox).

The cp4-epsps coding sequence is under control of a Figwort mosaic virus 34S promoter and a Pisum sativum rubisco small subunit terminator. A transit peptide from ribulose-1,5-bisphosphate carboxylase (Rubisco) small subunit was included before the cp4-epsps coding sequence and contributes to a higher level of expression in leaf tissues.

Note:
- The genetic element sizes were not readily available at the time of this record's creation.
- The gox coding sequence differs from the wild-type version of the gene at 3 amino acid sites (G85S, R153K and R334H) and was designated as goxv247.
- The cp4-epsps sequence was optimized for expression in plants.
- PCR and southern blot analyses indicated that the parental genome contains a single insertion event containing one copy of the T-DNA from plasmid PV-BNGT04. No genetic elements from outside of the right and left borders of the plasmid were transferred into or are present in the genomic DNA of the LMO.


For more information, kindly refer to the parental LMO records.
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LMO characteristics
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  • Food
  • Feed
Additional Information
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Records referencing this document Show in search
Record type Field Record(s)
Country's Decision or any other Communication Living modified organism(s) 1
Risk Assessment generated by a regulatory process Living modified organism(s) 1