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National Node of the BCH
Modified Organism
ACS-BNØØ5-8 x ACS-BNØØ3-6 - InVigor™ canola
Record information and status
Record ID
14760
Status
Published
Date of creation
2006-06-05 14:39 UTC (kirsty.mclean.consultant@cbd.int)
Date of last update
2013-07-23 20:24 UTC (dina.abdelhakim@cbd.int)
Date of publication
2013-07-23 20:24 UTC (dina.abdelhakim@cbd.int)

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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
EN
InVigor™ canola
Transformation event
MS8 x RF3
Unique identifier
ACS-BNØØ5-8 x ACS-BNØØ3-6
Developer(s)
Description
EN
The stacked canola line ACS-BNØØ5-8 x ACS-BNØØ3-6 was obtained through the conventional cross breeding of each of the parental organisms.  This results in a line with male-sterility, fertility restoration, pollination control system displaying glufosinate herbicide tolerance
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 rapa - Canola plant
ACS-BNØØ5-8 - InVigor™ canola
Changes in physiology and/or production - Reproduction - Male sterility Resistance to herbicides - Glufosinate
Show detection method(s)
ACS-BNØØ3-6 - InVigor™ canola
Changes in physiology and/or production - Fertility restoration Resistance to herbicides - Glufosinate
Show detection method(s)
Characteristics of the transformation process
Vector
EN
pTHW107 and pTHW118
Techniques used for the modification
  • Cross breeding
Genetic elements construct
 
pTA29 pollen specific promoter
1.51 Kb
 
 
Barnase
0.34 Kb
 
 
Barnase 3' Untranslated region
0.11 Kb
 
 
Nopaline Synthase Gene Terminator
0.26 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
 
Further details
Notes regarding the genetic elements introduced or modified in this LMO
EN
DNA insert from ACS-BNØØ5-8 vector pTHW107
ACS-BNØØ5-8 is a male-sterile canola line that cannot produce viable pollen due to the presence of the barnase gene. The line also contributes a copy of the bar gene which confers tolerance to the herbicide glufosinate.

DNA insert from ACS-BNØØ3-6 vector pTHW118
ACS-BNØØ3-6 is a fertility restorer canola line that inhibits the action of the barnase ribonuclease through the expression of the barstar gene. The line also contributes a copy of the bar gene which confers tolerance to the herbicide glufosinate.

For additional information on this LMO, please refer to the records of the parental LMOs.
LMO characteristics
Modified traits
Common use(s)
  • Food
  • Feed
Additional Information
Additional Information
EN
The canola lines MS8 and RF3 were developed using genetic engineering techniques to provide a pollination control system for the production of hybrid oilseed rape (MS8xRF3) expressing male sterility and tolerance to glufosinate ammonium. The novel hybridization system involves the use of two parental lines, a male sterile line MS8 and a fertility restorer line RF3. The transgenic MS8 plants do not produce viable pollen grains and cannot self-pollinate. In order to completely restore fertility in the hybrid progeny, line MS8 must be pollinated by a modified plant containing a fertility restorer gene, such as line RF3. The resultant F1 hybrid seed, derived from the cross between MS8 x RF3, generates hybrid plants that produce pollen and are completely fertile.

The male-sterile trait was introduced in MS8 by inserting the barnase gene, isolated from Bacillus amyloliquefaciens, a common soil bacterium that is frequently used as a source for industrial enzymes. The barnase gene encodes for a ribonuclease enzyme (RNAse) that is expressed only in the tapetum cells of the pollen sac during anther development. The RNAse affects RNA production, disrupting normal cell functioning and arresting early anther development, thus leading to male sterility.

The transgenic line RF3 was produced by genetically engineering plants to restore fertility in the hybrid line. Transgenic RF3 plants contain the barstar gene, also isolated from Bacillus amyloliquefaciens. The barstar gene codes for a ribonuclease inhibitor (barstar enzyme) expressed only in the tapetum cells of the pollen sac during anther development. The ribonuclease inhibitor (barstar enzyme) specifically inhibits barnase RNAse expressed by the MS8 line. Together, the RNAse and the ribonuclease inhibitor form a very stable one-to-one complex, in which the RNAse is inactivated. As a result, when pollen from the restorer line RF3 is crossed to the male sterile line MS8, the resultant progeny express the RNAse inhibitor in the tapetum cells of the anthers, allowing hybrid plants to develop normal anthers and restoring fertility.

Both transgenic lines MS8 and RF3 were also engineered to express tolerance to glufosinate ammonium, the active ingredient in phosphinothricin herbicides (Basta®, Rely®, Finale®, and Liberty®). Glufosinate chemically resembles the amino acid glutamate and acts to inhibit an enzyme, called glutamine synthetase, which is involved in the synthesis of glutamine. Essentially, glufosinate acts enough like glutamate, the molecule used by glutamine synthetase to make glutamine, that it blocks the enzyme's usual activity. Glutamine synthetase is also involved in ammonia detoxification. The action of glufosinate results in reduced glutamine levels and a corresponding increase in concentrations of ammonia in plant tissues, leading to cell membrane disruption and cessation of photosynthesis resulting in plant withering and death.
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