Journal of Environmental Biology

Genetic purity assessment of indica rice hybrids through DNA fingerprinting and grow out test

R.L. Verma¹*, S. Singh²,? P. Singh³, V. Kumar², S.P. Singh¹, S. Singh⁴, S. Samantaray¹ and O.N. Singh¹

¹Division of Crop Improvement, Indian Council of Agriculture Research-National Rice Research Institute, Cuttack-753 006, India

²Department of Genetics and Plant Breeding, College of Agriculture, G. B. Pant University of Agriculture and Technology, Pantnagar-263 145, India

³Department of Plant Breeding and Genetics, Veer Kunwar Singh College of Agriculture, Dumraon (Bihar Agricultural University, Sabour), Buxar-802 136, India

⁴Department of Agricultural Botany, Udai Pratap Autonomous College, Varanasi-221 002, India

*Corresponding Author E-mail: ram.pantvarsity@gmail.com??????????

Key words

DNA fingerprinting

Genetic purity?

Grow out test

Indica rice hybrids

Microsatellite markers

Publication Data

Paper received : 13.08.2016?????????????????? Revised received : 09.02.2017??????????????????? ??Re-revised received : 20.02.2017 Accepted : 23.03.2017???????????

Abstract

Aim: Varietal identity and their purity assessment is essential to assert intellectual rights to breeder, helps seed testing and certification agencies to ensure pure seeds to the farmers, buyers, processors and consumers.

Methodology: In the present study, molecular characterization and genetic purity analysis of nine indica rice hybrids and respective 13 parental lines (22 genotypes) were done utilizing grow out test grow out test and micro-satellite markers based DNA fingerprinting approach. A total of 48 microsatellite (SSR and STMS) markers spanning over whole rice genome were screened. ????

Results: The statistical analysis of grow out test revealed that 88.25 per cent genetic purity was assessed in 3 hybrids tested. Twelve out of 48 microsatellite used were found polymorphic and amplified a total of 29 alleles among the genotypes with an average polymorphic information content value of 0.45. Out of 12 informative markers, nine were found polymorphic across the genotypes and produced unique fingerprints. Five STMS markers (RM154, RM164, RM234, RM258 and RM519) showed polymorphism over all the genotypes and were identified as referral marker for use in unambiguous identification and protection of nine hybrids. Nevertheless, marker RM 19 amplified allele was specific to the parental lines of CNRH 3 and APHR 2 (having common CMS line, IR62829). The results on genetic purity test through GOT had highly significant positive correlation (r=0.96) with molecular marker analysis. ????????

Interpretation: Genotype specific STMS markers identified in the study and their close association with the ability of grow out test, emphasizes immense scope for molecular approaches to be an alternatives in unambiguous assessment of genetic purity of hybrids and their parents. The molecular markers based genetic purity assessment may serve as a probable substitute to grow out test, which is a cumbersome and time consuming task and would benefit the farmers and agencies involved in this venture, hybrid rice seed production and cultivation.

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