“CHARACTERIZATION OF DROUGHT RESISTANT AND SUSCEPTIBLE PEARL MILLET (Pennisetumglaucum(L.) R.Br.) GENOTYPES USING BIOCHEMICAL AND MOLECULAR MARKERS”
Loading...
Files
Date
2015-09
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
jau,junagdh
Abstract
The present experiment on “CHARACTERIZATION OF DROUGHT RESISTANT AND SUSCEPTIBLE PEARL MILLET (Pennisetumglaucum(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 differentpearlmillet genotypes using biochemical techniques viz., Isoenzymes and to screen the drought susceptibility and suceptibilty analysis among pearlmillet genotypes using various molecular markers viz. RandomAmplified Polymorphic DNA (RAPD), Inter Simple Sequence Repeats (ISSR) and Simple Sequence Repeats (SSR).
The protein profile (Native-PAGE) produced eight bands with Rm values ranging from 0.031 to 0.659 while SDS-PAGE of pearlmilletseed produced total of twenty bands with Rm values ranging from 0.043 to 0.882. SDS-PAGE in present study showed specific bands linked with drought reaction trait in resistant genotype J-2454. The isoenzymic study of peroxidase revealed fourteen bands ofRm values were ranging from 0.029 to 0.721 whileisoenzymic study of super oxide dismutase revealed three bands ofRm values ranging from 0.253 to 0.619,whileisoenzymic study of poly phenol oxidase revealed eight bands ofRm values ranging from 0.123 to 0.742.The combined study of protein profiling and isoenzymes revealed that the phylogenetic tree consisted of two main clusters with similarity coefficient range 48 to 83%.
Total 20 RAPD primers generated 378 bands in which 273 bands were polymorphic with an average of 13.65bands per primer. The 7 ISSR primers were screened to generate 133bands in which 120 bands were polymorphic with an average of 17.14bands per primer.Total 56 SSR primers generated the 487 fragments in which 419 bands were polymorphic with an average of 7.48 bands per primer. Thesimilarity coefficientranged from 60 to 81% for RAPD, 34 to 68% for ISSR and 45 to 66% for SSR.The pooled study of RAPD, ISSR and SSR generated clustering pattern which seem to be similar as SSR clustering pattern.RAPD primers (OPA-18, OPC-05, OPD-03, OPD-05, OPM-14, OPN-13, OPO-09, OPO-15, OPQ-12, OPQ-14, OPR-14 and OPZ-10) was able to produce genotype specific band, while ISSR primers (ISSR-I, ISSR-II, ISSR-IV, ISSR-X and ISSR-25) and SSR primers (PSMP-2018, PSMP-2072, PSMP-2074, PSMP-2077, PSMP-2080, PSMP-2084, PSMP-2089, PSMP-2201, PSMP-2203, PSMP-2208, PSMP-2220, PSMP-2232, PSMP-2267, PSMP-2273, PSMP-2276, PSMP-2277, ICMP-3002, ICMP-3024, ICMP-3050, ICMP-3066, ICMP-3080, ICMP-3081, ICMP-3086, ICMP-3088, ICMP-4010, Xpsmp-2069, Xpsmp-2089, Xpsmp-2208, Xpsmp-2219, Xpsmp-2255, Xpsmp-2266, Xpsmp-2270, Xpsmp-2273, Xpsmp-2275, Xicmp-3027, Xicmp-3088 and Xctm-12) produced genotype specific bands and discriminated different genotypes among all 10 pearlmillet genotypes respectively.Among all molecular and biochemical study techniques, ISSR and SSR markers and SDS-PAGE seem to be more informative than RAPD, and other isoenzymeassay to access distinguish drought resistant and susceptible pearlmillet genotypes.
RAPD, ISSR and SSR study indicated that resistant genotype J-2467 is most diverse among 10 pearlmillet genotypes used in this study.
Thus, molecular markers were proved to be more accurate and effective than biochemical markers except SDS-PAGE and ISSR as some of RAPD and SSR primers found to be genotype specific markers in the present study.
Key words: Isoenzyme, RAPD, ISSR, SSR, Pennisetumglaucum.
Description
Keywords
BIOTECHNOLOGY