Dhara Mustard Hybrid-11

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.

Kindly refer to parental records for more information.

pPZP200

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.