“CHARACTERISATION OF DOWNY MILDEW RESISTANT AND SUSCEPTIBLE PEARL MILLET (Pennisetum glaucum (L.) R.Br.) GENOTYPES USING BIOCHEMICAL AND MOLECULAR MARKERS”

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Date
2014-05
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jau,junagadh
Abstract
The present experiment on “CHARACTERISATION OF DOWNY MILDEW RESISTANT AND SUSCEPTIBLE PEARL MILLET (Pennisetum glaucum (L.) R.Br.) GENOTYPES USING BIOCHEMICAL AND MOLECULAR MARKERS” was conducted at Department of Biotechnology, Junagadh Agricultural University, Junagadh with objectives, to detect the polymorphism among different pearlmillet genotypes using biochemical techniques viz., Isoenzymes and to screen the disease susceptibility and suceptibilty analysis among pearlmillet genotypes using various molecular markers viz. Random Amplified Polymorphic DNA (RAPD), Inter Simple Sequence Repeats (ISSR) and Simple Sequence Repeats (SSR). The protein profile (Native-PAGE) produced thirteen bands with Rm values ranging from 0.176 to 0.950 while SDS-PAGE of pearlmillet seed produced total of seventeen bands with Rm values ranging from 0.041 to 0.899. The isoenzymic study of peroxidase revealed seven bands of Rm values were ranging from 0.262 to 0.840 while isoenzymic study of super oxide dismutase revealed three bands of Rm values ranging from 0.256 to 0.622,while isoenzymic study of poly phenol oxidase revealed five bands of Rm values ranging from 0.120 to 0.430. The combined study of protein profiling and isoenzymes revealed that the phylogenetic tree consisted of two main clusters with similarity coefficient range 78 to 97%. Total 10 RAPD primers generated 118 bands in which 105 bands were polymorphic with an average of 10.5 bands per primer. The 9 ISSR primers were screened to generate 120 bands in which 79 bands were polymorphic with an average of 7.90 bands per primer. Total 80 SSR primers generated the 244 fragments in which 169 bands were polymorphic with an average of 2.1 bands per primer. The similarity coefficient ranged from 44 to 79% for RAPD, 62 to 64% for ISSR and 58 to 72% for SSR. The pooled study of RAPD, ISSR and SSR generated clustering pattern which seem to be similar as SSR clustering pattern. No RAPD primers was able to produce genotype specific band, while ISSR primers (ISSR-V, ISSR-VIII and ISSR-IX) and SSR primers (PSMP-2030, PSMP-2073, PSMP-2072, PSMP-2077, PSMP-2225, PSMP-2070, PSMP-2084, PSMP-2202, PSMP-2208, PSMP-2274, PSMP-2018, PSMP-2224, ICMP-3013, ICMP-3017, ICMP-3024, ICMP-3057, Xicmp-3088, ICMP-4014, Xpsmp-2255, Xpsmp-2248, Xpsmp-2249, Xpsmp-2270, Xpsmp-2227, m 13 Xpsmp-2237, Xpsmp-2273, Xpsmp-2269) produced genotype specific bands and discriminated different genotypes among all 10 pearlmillet genotypes respectively. Among all molecular and biochemical study techniques, SSR markers ans SDS-PAGE seem to be more informative than RAPD, ISSR and other isoenzyme assay to access distinguish disease resistant and suceptible pearlmillet genotypes. Thus, molecular markers were proved to be more accurate and effective than biochemical markers exept SDS-PAGE as some of RAPD, ISSR and SSR primers found to be genotype specific markers in the present study. Key words: Isoenzyme, RAPD, ISSR, SSR, Pennisetum glaucum.
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biotechnology
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