BIOCHEMICAL AND MOLECULAR ANALYSIS OF DROUGHT TOLERANT AND SUSCEPTIBLE CULTIVARS OF PEARL MILLET [Pennicetum glaucum L.)
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Date
2007-05-11
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MAHATMA PHULE KRISHI VIDYAPEETH RAHURI-413 722, DIST. AHMEDNAGAR (MS) INDIA
Abstract
The present investigation was carried out to study the proline metabolism in response to osmotic stress and RAPD analysis of the genomic DNA from 4 drought tolerant and 2 drought susceptible cultivars of pearl millet with 18 random decamer primers. The seeds of 16 cultivars comprising of tolerant and susceptible hybrids, R lines and local germplasm were germinated on petnplates in duplicate sets for 5 days under ambient conditions. An osmotic stress of-1.0 MPa was applied to seedlings m one set with PEG-6000 solution, roots were collected after 5 days and analyzed for changes in proline content, P5CS activity and soluble proteins. The genomic DNA from 2 susceptible and 4 tolerant hybrids was extracted from 15 days old seedlings grown in earthern pots. Then purified DNA was subjected to PCR mplification with 18 random decamerprimers XV Abstract contd.... Miss. Maid S.M.Application of osmotic stress was found to increasethe proline accumulation in root tissues. Shradha cultivar had the highest proline accumulation (17.55 pinoles g1 fr.wt). The P5CS activity was found to increase under stressed condition. Shradha cultivar had the highest P5CS activity i.e. 0.50 pmole-yglutamyl hydroxamate g1 mm-1. Similarly application of osmotic stress was found to increase the root soluble proteins. Shradha cultivar had also the highest accumulation of soluble protein content. The result indicated that an increased proline accumulation due to stress is a good indicator of drought tolerance in pearl millet. An increased P5CS activity during stress indicated that fresh biosynthesis of proline in roots during stress supported the increased enzyme synthesis and the least protein degradation during the stress. "yC p 3^> '• RAPD analysis of genomic DNA of six cultivars revealed 37.50 to 88.89 % olymorphism with 18 random primers. Among the primers tested OPC-20 showed a distinct amplicon of 0.34 kb size and OPA-12 exhibited distinct amphconof 0.52 kb size in tolerant cultivars. These distinct bands can be used in designing specific primers for further screening of segregating populations of pearl millet .
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sowing, intercropping, wheats, tillage equipment, seed drilling, crops, mustard, fertilizers, land resources, chickpeas