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Use of Molecular and Protein Based Markers for Accessing Genetic Diversity Among Hippophae salicifolia D. Don Genotypes of Uttarakhand

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Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

In order to study the genetic diversity among 16 genotypes of Hippophae salicifolia D. Don of Uttarakhand region, random amplified polymorphic DNA (RAPD)–PCR analysis along with protein profiling of seed storage proteins was carried out for selection of superior genotype having high economic importance. A total of 21 RAPD primers were assayed for their specificity in detecting genetic variability in H. salicifolia genotypes, of which 11 RAPD primers were highly reproducible and were found suitable for use in PCR analysis. A total of 198 bands were scored corresponding to an average of 10.8 bands per primer with 138 bands showing polymorphism (73.2 %) with similarity coefficient ranging from 0.011 to 0.999. A dendrogram constructed based on the UPGMA clustering method revealed two major clusters. Cluster-I comprises of 13 cultivars, while cluster-II includes the remaining three cultivars. The cultivar ST-4, ST-5 and ST-16 were quite unique from the remaining cultivars as evident from the dendrogram. The analysis of total seed storage protein (quantitative) and protein profiling (qualitative) among 16 genotypes of H. salicifolia was performed to indentify novel proteins of important functional attributes. 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. This study helps in the selection of superior genotype of H. salicifolia having higher economic importance by using developed molecular and protein based markers in Uttarakhand regions.

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Abbreviations

RAPD:

Random amplified polymorphic DNA

PCR:

Polymerase chain reaction

SDS:

Sodium do-decyl sulphate

PAGE:

Polyacrylamide gel electrophoresis

SBT:

Seabuckthorn

PIC:

Polymorphic information content

UPGMA:

Unweighted pair group method with arithmetic mean

AFLP:

Amplified fragment length polymorphism

TBSP:

Total buffer soluble protein

SSR:

Simple sequence repeats

SNPs:

Single nucleotide polymorphism

DArT:

Diversity arrays technology

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Acknowledgments

The authors acknowledge the Dean, Hill campus, G.B. Pant Agriculture and Technology Ranichauri for providing seed materials. They also thank to Dr. V.K. Yadav, Hill campus Ranichauri, G.B. Pant Agriculture and Technology for providing the passport in formation of H. salicifolia genotype used into the present work.

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Authors and Affiliations

  1. Department of Molecular Biology & Genetic Engineering, College of Basic Sciences & Humanities, G. B. Pant University of Agriculture & Technology, U. S. Nagar, Pantnagar, 263 145, Uttarakhand, India

    Netrapal Sharma, Sadhna Singh & Anil Kumar

  2. Department of Biotechnology, SBS PG Institute of Biomedical Sciences, Balawala, Dehradun, Uttarakhand, India

    Sanjay Gupta

  3. Defence Institute of Bio-Energy Research, Defense Research & Development Organization, Goraparao, Nainital, Haldwani, Uttarakhand, India

    Sanjay Mohan Gupta

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  1. Netrapal Sharma
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  2. Sanjay Gupta
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  3. Sadhna Singh
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  4. Sanjay Mohan Gupta
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  5. Anil Kumar
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Correspondence to Anil Kumar.

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Sharma, N., Gupta, S., Singh, S. et al. Use of Molecular and Protein Based Markers for Accessing Genetic Diversity Among Hippophae salicifolia D. Don Genotypes of Uttarakhand. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 84, 47–54 (2014). https://doi.org/10.1007/s40011-013-0196-4

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  • DOI: https://doi.org/10.1007/s40011-013-0196-4

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