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Modified Organism
BCS-BNØ12-7 × ACS-BNØØ3-6 × MON-883Ø2-9 - Fertility restored, Herbicide tolerant canola
Record information and status
Record ID
115686
Status
Published
Date of creation
2020-08-25 19:25 UTC (austein.mcloughlin@cbd.int)
Date of publication
2020-08-25 19:25 UTC (austein.mcloughlin@cbd.int)

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.

LMO name
Fertility restored, Herbicide tolerant canola
Transformation event
MS11 × RF3 × MON88302
Unique identifier
BCS-BNØ12-7 × ACS-BNØØ3-6 × MON-883Ø2-9
Developer(s)
Description
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.
Recipient Organism or Parental Organisms
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.
Brassica napus - Turnip, Rapeseed, Canola Plant, Oilseed Rape, Rape, BRANA
BCS-BNØ12-7 - Male Sterile/ Fertility Restored Herbicide Tolerant Canola
Changes in physiology and/or production - Reproduction - Male sterility Resistance to herbicides - Glufosinate
ACS-BNØØ3-6 - InVigor™ canola
Changes in physiology and/or production - Fertility restoration Resistance to herbicides - Glufosinate
Show detection method(s)
MON-883Ø2-9 - TruFlex Roundup Ready™ Canola
Resistance to herbicides - Glyphosate
Show detection method(s)
Point of collection or acquisition of the recipient organism
Please note the developer has not been confirmed.
Characteristics of the transformation process
Vector
pTCO113; pTHW118; PV-BNHT2672
Techniques used for the modification
  • Cross breeding
Genetic elements construct
 
Transcript 7 gene 3' untranslated region
0.21 Kb
 
 
Phosphinothricin N-acetyltransferase gene
0.55 Kb
 
 
rbcS Promoter
1.73 Kb
 
 
Nopaline Synthase Gene Terminator
0.26 Kb
 
 
Barnase 3' Untranslated region
0.10 Kb
 
 
Barnase
0.34 Kb
 
 
pTA29 pollen specific promoter
1.51 Kb
 
 
Nopaline Synthase Gene Promoter
0.29 Kb
 
 
Barstar
0.27 Kb
 
 
Transcript 7 gene 3' untranslated region
0.21 Kb
 
 
pTA29 pollen specific promoter
1.51 Kb
 
 
Barstar
0.27 Kb
 
 
Barstar gene terminator
0.04 Kb
 
 
Nopaline Synthase Gene Terminator
0.26 Kb
 
 
rbcS Promoter
1.73 Kb
 
 
Phosphinothricin N-acetyltransferase gene
0.55 Kb
 
 
Transcript 7 gene 3' untranslated region
0.21 Kb
 
 
FMV 35S Enhancer
1.04 Kb
 
 
Elongation factor EF-1alpha promoter
0.00 Kb
 
 
Elongation factor EF-1alpha Leader
0.05 Kb
 
 
Elongation factor EF-1alpha Intron 1
0.62 Kb
 
 
Chloroplast transit peptide 2
0.23 Kb
 
 
5-enolpyruvylshikimate-3-phosphate synthase gene
1.37 Kb
 
 
rbcS-E9 gene terminator
0.64 Kb
 
Further details
Notes regarding the genetic elements introduced or modified in this LMO
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.
LMO characteristics
Modified traits
  • Fertility restoration
Common use(s)
  • Food
  • Feed
Additional Information
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.

Records referencing this document (2)
IDDescription
2record(s) found
Country's Decision or any other Communication1 record
Risk Assessment1 record