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Modified Organism
Male sterile Indian mustard
Record information and status
Record ID
115909
Status
Published
Date of creation
2021-02-23 16:58 UTC (austein.mcloughlin@cbd.int)
Date of last update
2021-02-23 17:25 UTC (austein.mcloughlin@cbd.int)
Date of publication
2021-02-23 17:25 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
Male sterile Indian mustard
Transformation event
bn 3.6
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 male-sterility and herbicide tolerance. For male-sterility, the mustard expresses Bacillus amyloliquefaciens barnase, an RNase, in the tapetum cell layer of the pollen sac during anther development. Expression of the non-specific RNase, causes RNA to be degraded and leads to the failure of pollen development. Since female fertility is not expected to be impacted, this line can be used in the production of hybrid seeds because the plants cannot self-pollinate and thus require pollen from another parent.

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

The mustard variety RLM 198 was initially transformed with the barnase construct and then backcrossed into variety Varuna to achieve the parental line Varuna bn 3.6 for hybrid seed production. 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
RLM 198 was developed through mutation and recombination breeding by Punjab Agricultural University. Varuna (T-59) is extensively cultivated in northern India.
Characteristics of the transformation process
Vector
pPZP200
Techniques used for the modification
  • Agrobacterium-mediated DNA transfer
Genetic elements construct
 
CaMV 35S promoter
0.00 Kb
 
 
5' Untranslated Leader of AMV RNA4
0.00 Kb
 
 
Phosphinothricin N-acetyltransferase gene
0.00 Kb
 
 
Octopine Synthase Gene Terminator
0.00 Kb
 
 
Topoisomerase
3.00 Kb
 
 
Acetohydroxy acid synthase gene
2.00 Kb
 
 
pTA29 pollen specific promoter
0.87 Kb
 
 
Barnase
0.33 Kb
 
 
CaMV 35S terminator
0.00 Kb
 
Further details
Notes regarding the genetic elements introduced or modified in this LMO
The modified brown mustard contains two gene cassettes: Streptomyces hygroscopicus phosphinothricin N-acetyltransferase (bar) and Bacillus amyloliquefaciens barnase.

The bar coding sequence is under control of a Cauliflower mosaic virus (CaMV) 35S promoter with an Alflafa mosaic virus (AMV) leader and an Rhizobium radiobacter octopine synthase gene terminator. The AMV leader sequence enhances expression of bar.

Barnase is under control of a Nicotiana tabacum TA29 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).

A spacer fragment can be found between the two gene cassettes to prevent 'leaky' expression of barnase from the CaMV promoter. It is comprised of Pisum sativum topoisomerase (3kb) and Arabidopsis thaliana acetohydroxy acid synthase (2 kb) fragments. Each fragment contains truncations on the 3' and 5' ends. No open reading frames were created during the fusion of the two fragments and thus are not expected to encode a functional product. Note: The arrangement of the insert is unclear.

Note:
- Southern blot and segregation analyses indicated that the genome contains a single insertion.
- Sequencing analysis indicated that the T-DNA integrated was identical to the sequences in the vector.
- The T-DNA was found to be integrated in A9 Linkage Group on the 'A' genome between Bra32488 and Bra32489 genes.
LMO characteristics
Modified traits
Common use(s)
  • Hybrid seed production

Records referencing this document (1)
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