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Aim: Millets are a group of highly variable small seeded grasses, grown around the world as cereal crops or grains for fodder and human food. In this study, screening of crop primers with 8 genotypes of millet crop species belonging to genus Panicum, Eleusine, Echinochloa and Brachiaria was done. Main objective was to understand the genetic diversity among 7 millet crops varieties and a wild grass using RAPD markers by targeting nuclear gene. To find out the relationship between species of the same genus based on RAPD markers and to construct phylogenetic tree using PyElph 1.4 and Past326b softwares.
Methodology: PCR analysis was done using the selected RAPD to identify the polymorphic loci between the 8 genotypes. Primers OPC06, OPD13, OPW04, OPC18 were selected for analysis based on the band patterns, for further RAPD analysis. Bar Coding and Sequencing was done for validation of species with the help of softwares BLAST and Clustal OMEGA. Phylogenetic tree was constructed using all 4 primers with the help of software tools PyElph 1.4 and Past326b. The point parameter, which guided the joining linkage rule, is unweighted pair group average (UPGMA) and the genetic distance was computed from raw data.
Outcome: Among the 4 primers used, OPW04 and OPD13 showed maximum scoring of bands based on the phylogeny were established between the crop species of Eleusine species and the Indian Goose grass. The dendrogram showed that the varieties could be grouped into clusters. The sample C (Elusine indica), D (Elusine coracana) is diverged from the same root and C, D and G (Indian goose grass) are from the same root. Sample G is found to be very closely related to the C and D.
Agrama HA, Tunistra MR. Phylogenetic diversity and relationships among Sorghum accessions using SSRs and RAPDs. Department of Agronomy. Kansas state university, Throckmorton, Manhattan, USA; 2004.
Gopalan B, Ramasastri SC, Balasubramanian. Nutritive value of Indian foods Hyderabad, India: National institute of Nutrition, Indian council for medical research; 1976.
Goodarzi F, et al. Genetic analysis of castor (Ricinus communis L.) using ISSR markers. J Plant Mole Breeding. 2015; 3(1):18-34.
Sharma MV, Kantartzi SK, Stewart JM. Molecular diversity and polymorphism information content of selected Gossypium hirsutum accessions. Summ Ark Cotton Res. 2009;124–126.
Ravi M, Geetanjali S, Sameeya Farheen F, Maheswaram M. Molecular Marker based genetic diversity analysis in rice (Oryza sativa L) using RAPD and SSR markers. Euphytica. 2003;133:243-252
Kajal Kumari, Mehanathan Mutha-milarasan, Gopal Misra, Sarika Gupta. Development of eSSR-markers in Setaria italica and their applicability in studying genetic diversity, cross transferability and comparative mapping in millet and non-millet species. National Institute of Plant Genome Research, New Delhi, India; 2013.
Neelam Tiwari, Sharad Tiwari, Niraj Tripathi. Genetic characterization of Indian little millet (Panicum sumatrense) genotypes using random amplified polymorphic DNA markers; 2018.
Jeff J. Doyle and Jane L. Doyle. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin. 1987;19(1):11-15.
yvind Hammer, David A, Harper T, Paul D. Ryan, 2001 PAST: Paleontological statistics software package for education and data analysis. Palaeontologia Electronica. 2001;4(1), Part. 4:9pp, 178kb.
Madho Singh Bisht, Yasuhiko Mukai. Mapping of rDNA on the chromosomes of Eleusine species by fluorescence in situ hybridization. Genes & Genetic Systems. 2001;75(6):343-8.
Salimath SS, de Oliveira AC, Godwin ID, Bennetzen JL. Assessment of genome origins and genetic diversity in the genus Eleusine with DNA markers. Genome. 1995;38:757–763.
Tiwari KK, Singh A, Pattnaik S, Sandhu M, Kaur S, Jain S, Tiwari S, Mehrotra S, Anumalla M, Samal R, Bhardwaj J. Identification of a diverse mini-core panel of Indian rice germplasm based on genotyping using microsatellite markers. Plant Breed. 2015;134(2):164–171.
Sruthi K, Divya B, Senguttuvel P, Revathi P, Kemparaju KB, Koteswara Rao P, Sundaram RM, Vikram Jeet Singh, Ranjith Kumar E, Prolay Kumar Bhowmick, Vinod KK, Gopala Krishnan S, Singh AK, Hari Prasad AS. Evaluation of genetic diversity of parental lines for development of heterotic groups in hybrid rice (Oryza sativa L). Journal of Plant Biochemistry and Biotechnology. 2020;29: 236–252
Ning Yu, Yu-long Wei, Xin Zhang, Ning Zhu, Yan-li Wang, Yue Zhu, Hai-ping Zhang, Fen-mei Li, Lan Yang, Jia-qi Sun, Ai-dong Sun. Barcode ITS2: A useful tool for identifying Trachelospermum jasminoides and a good monitor for medicine market; 2017
Rachie KO. The systematic collection of sorghums, millets and maize in India. Report of the Rockefeller Foundation and the Indian Council of Agricultural Research, New Delhi; 1965.