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PhD work |
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I did my PhD work in NRCPB, Indian Agricultural Research Institute (IARI), New Delhi, India.
I have submitted my thesis to Department of Biotechnology, Ch. Ch. Singh University, Meerut, India.
Chairman: Prof. P. C. Sharma.
Co-chairman: Dr. P. A. Kumar. |
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My thesis work is on "Genetic transformation and DNA fingerprinting in pigeonpea (Cajanus cajan)". In this I have done the following objectives.
1. Development of drought tolerance pigeonpea plant by using Mannito-dehydrogenase (MtlD) gene through Agrobacterium transformation method.
2. DNA fingerprinting of cultivated and wild species of pigeonpea by using AFLP.
3. Isolation and characterization of green tissue-specific promoter (rbcS) from pigeonpea.
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Pigeonpea (Cajanus cajan) is one of the major legume crops of the tropic and sub-tropics, yielding nutritious protein rich seeds for human consumption and animal feed. Being highly recalcitrant to regeneration, among all the plant species, tropical grain legumes are difficult for genetic transformation. Improvement of legumes by genetic engineering requires procedure to regenerate them in vitro. Studies have been conducted to find out the efficient regeneration protocol for genetic transformation in pigeonpea in the present investigation. By using Mannito-dehydrogenase (MtlD) gene, drought tolerance transgenic pigeonpea was developed by using Agrobacterium mediated transformation. A green tissue-specific promoter (rbcS) from pigeonpea was isolated through PCR by using the primers of pea rbcS3A promoter. The pigeonpea promoter was sequenced and compared with the pea promoter for the sequence homology, light-indusibility, tissue-specificity and stage-specificity by expressing GUS-reporter gene in transgenic tobacco.
Wild relatives of pigeonpea serves as rich source of disease resistance genes that can be introgressed into the cultivated species. However, development of improved types through recombinations and hybridizations have met with limited success in pigeonpea and other species of Cajanus. Research efforts need to develop a good understanding of genetic systems controlling qualitative and quantitative characters. Genetic diversity of various cultivated species of pigeonpea and two of its wild species (Cajanus volubilis and Cajanus bracteata) were studies by using PCR based markers like AFLP, RAPD and ISSR's. The studies were also conducted to findout the efficient and low-cost markers in pigeonpea. |
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