Advances in Animal and Veterinary Sciences

Research Article
Adv. Anim. Vet. Sci. 3 (4): 211 - 224
http://dx.doi.org/10.14737/journal.aavs/2015/3.4.211.224
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Netrapal Sharma1, 3*, Sanjay Gupta2, Rekha Pathak3, Sanjay Mohan Gupta4, Tserin Stobdan5, Anil Kumar1

1Department of Molecular Biology & Genetic Engineering, G. B. Pant University of Agriculture & Technology, Pantnagar- 263 145; 2Division of Surgery, Indian Veterinary Research Institute, Izatnagar, Bareilly-243122, U.P.; 3Department of Biotechnology, SBS PG Institute of Biomedical Sciences, Balawala, Dehradun; 4Defence Institute of Bio-Energy Research, DRDO, Haldwani, 263139; 5Defence Institute of High Altitude Research, DRDO, Leh-Ladakh, India.

*Correspondence | Netrapal Sharma, Indian Veterinary research Institute, Izatnagar, U.P., India; Email: np_biotech53@rediffmail.com

Abstract
The genetic diversity among thirty six genotypes of Hippophae Salicifolia D. Don of Uttarakhand region and Hippophae rhamnoides subsp. Turkestanica of Laddakh region was studied; using simple sequence repeats DNA (SSRs)-PCR analysis. The protein profiling and antioxidant profiling (quantitative and qualitative) of seeds was also done to select the superior genotypes having high economic importance. A total of seven simple sequences repeat primers used for detecting genetic variability in Sea buck thorn seeds out of which Seven SSR primers were highly reproducible and found suitable for use in PCR (Polymerase Chain Reaction) analysis. SSRs profile obtained by primer UTR-15 was containing total of 8 bands (0.15- 0.6 kb), among these eight bands, one was found to be polymorphic (12.5%). The similarity coefficients based on SSR markers ranged from 0.011 to 0.999 with an average value of 0.226. The polymorphism information content (PIC) value ranged from 0.1686 (UTR-015) to 0.2975 (UTR-12) with a mean of 0.351. A dendrogram constructed based on the Unweighted Pair Group Method with Arithmetic Mean (UPGMA) clustering method revealed two major clusters of Uttarakhand (with subclusters) and Laddakh region (no subclusters). The analysis of total seed storage protein (quantitative) and protein profiling (qualitative) among the genotypes was performed to indentify novel proteins of important functional attributes. Sodium dodecayl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) based generated profiles of seed proteins showed major differences in banding patterns among these genotypes with respect to altitude besides differences in seed protein contents. The phenol contents of the H. salicifolia (Particularly Genotype ST 10) was found significantly higher (P<0.05) than H. rhamnoides (Genotype DIHAR-6). The flavonoid content of the DIHAR-14 was higher as compared to ST-1. Total scavenging antioxidant was quite high and equally good in all the genotypes. This study helps in the selection of superior genotypes having higher nutraceutical value by using developed molecular and protein based markers in Uttarakhand and Laddakh regions.

Keywords | Sea buckthorn, Simple sequence repeats, Genetic diversity, Anti-oxidant activity