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Fertility restorer Indian mustard
University of Delhi
Centre for Genetic Manipulation of Crop Plants (CGMCP)
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.
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
Varuna (T-59) is extensively cultivated in northern India. EH-2 was
developed from east European material by University of Nagpur.
- Agrobacterium-mediated DNA transfer
Octopine Synthase Gene Terminator
Phosphinothricin N-acetyltransferase gene
CaMV Enhanced 35S promoter
pTA29 pollen specific promoter
The modified mustard contains two gene cassettes: Streptomyces
hygroscopicus (bar) and Bacillus
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.
- 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.
- Changes in physiology and/or production
- Resistance to herbicides