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Modified Organism
Fertility restorer Indian mustard
Record information and status
Record ID
115910
Status
Published
Date of creation
2021-02-23 17:07 UTC (austein.mcloughlin@cbd.int)
Date of publication
2021-02-23 17:07 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 restorer Indian mustard
Transformation event
modbs 2.99
Developer(s)
University of Delhi
Centre for Genetic Manipulation of Crop Plants (CGMCP)
New Delhi
India
Url:University of Delhi Centre for Genetic Manipulation of Crop Plants,Contact information
Description
The Indian mustard (Brassica juncea) was modified for restoring male fertility and herbicide tolerance. For male fertility restoration, the mustard expresses Bacillus amyloliquefaciens barstar, a protein inhibitor of barnase, in the tapetum cell layer of the pollen sac during anther development. Barnase is an RNase, which degrades RNA in the pollen and prevents the pollen from developing. With barstar expression, the RNase is inhibited and the pollen develops normally. Thus, crossing barstar expressing lines with barnase expressing (male sterile) lines restores male fertility in the progeny.

For glufosinate tolerance, the mustard expresses Streptomyces hygroscopicus phosphinothricin N-acetyltransferase, which inactivates the herbicide through acetylation.

To achieve the fertility restorer line EH-2 modbs 2.99 for hybrid seed production, the mustard variety Varuna was initially transformed with transgenic construct and then backcrossed into variety EH-2. Six backcrossings were performed to try to remove integration of vector backbone sequence.
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 juncea - Indian mustard, Brown mustard, Chinese mustard, Leaf mustard, Vegetable mustard, Mustard greens, BRAJU
Point of collection or acquisition of the recipient organism
Varuna (T-59) is extensively cultivated in northern India. EH-2 was developed from east European material by University of Nagpur.
Characteristics of the transformation process
Vector
pPZP200
Techniques used for the modification
  • Agrobacterium-mediated DNA transfer
Genetic elements construct
 
Octopine Synthase Gene Terminator
0.00 Kb
 
 
Phosphinothricin N-acetyltransferase gene
0.00 Kb
 
 
CaMV Enhanced 35S promoter
0.00 Kb
 
 
pTA29 pollen specific promoter
0.28 Kb
 
 
Barstar
0.27 Kb
 
 
CaMV 35S terminator
0.00 Kb
 
Further details
Notes regarding the genetic elements introduced or modified in this LMO
The modified mustard contains two gene cassettes: Streptomyces hygroscopicus (bar) and Bacillus amyloliquefaciens barstar.

The bar coding sequence is under control of the Cauliflower mosaic virus (CaMV) enhanced 35S promoter and Rhizobium radiobacter octopine synthase terminator.

Barstar is under control of a Nicotiana tabacum TA29 pollen specific promoter and a CaMV 35S terminator. The TA29 promoter is active only in the tapetum cell layer of the pollen sac during anther development (male-specific expression). The coding sequence was codon optimized for expression in plants.

Note:
- Southern blot and segregation analyses indicated that the genome contains a single insertion
- DNA sequence analysis indicated that the insertion occurred in the 'B' genome.
LMO characteristics
Modified traits
Common use(s)
  • Hybrid seed production

Records referencing this document (1)
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