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Modified Organism
Dhara Mustard Hybrid-11
Record information and status
Record ID
115911
Status
Published
Date of creation
2021-02-23 17:30 UTC (austein.mcloughlin@cbd.int)
Date of publication
2021-02-23 17:30 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
Dhara Mustard Hybrid-11
Transformation event
DMH-11 (Varuna bn 3.6 × EH-2 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 hybrid DMH-11 Indian mustard (Brassica juncea) was produced through crossing two modified parental lines for restored male-fertility and herbicide tolerance. The mustard expresses expresses both Bacillus amyloliquefaciens barnase, an RNase, and barstar, an inhibitor of barnase, in the tapetum cell layer of the pollen sac during anther development. Without barstar, the non-specific nature of barnase would degrade the RNA and prevent the pollen from developing. The expression of barnase allowed the developers to control crosses of the parental plants. In this specific cross, fertility (pollen production) was restored through the introduction of barstar. Overall, the resulting heterosis is expected to improve productivity of the hybrid mustard.

For glufosinate tolerance, the mustard expresses Streptomyces hygroscopicus phosphinothricin N-acetyltransferase, which inactivates the herbicide through acetylation.
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
Male sterile Indian mustard
Changes in physiology and/or production - Reproduction - Male sterility Resistance to herbicides - Glufosinate, Imidazolinone, Sulfonylurea
Fertility restorer Indian mustard
Changes in physiology and/or production - Fertility restoration Resistance to herbicides - Glufosinate
Point of collection or acquisition of the recipient organism
Kindly refer to parental records for more information.
Characteristics of the transformation process
Vector
pPZP200
Techniques used for the modification
  • Cross breeding
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
 
 
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
Insertion related to Varuna bn 3.6
The parental genome 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.


Insertion related to EH-2 modbs 2.99
The parental genome contains two gene cassettes: (bar) and B. amyloliquefaciens barstar.

The bar coding sequence is under control of a CaMV enhanced 35S promoter and R. radiobacter octopine synthase terminator.

Barstar is under control of a N. 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.


For more information, kindly refer to the parental records.
LMO characteristics
Modified traits
Common use(s)
  • Food
Additional Information
Other relevant website address or attached documents