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Development of EST-SSR markers for genetic diversity analysis in coconut (Cocos nucifera L.)

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Abstract

Genetic improvement in coconut relies on exploiting the vast existing diversity among coconut accessions. Robust molecular markers are a pre-requisite for efficient characterization of genetic diversity. Microsatellites or simple sequence repeats (SSRs), mined from expressed sequence tags (ESTs), constitute an important resource for analysis of genetic diversity as they are abundant, polymorphic and represent function regions of the genome. We have identified a total of 318,528 putative EST-SSRs from 130,942 unigenes utilizing a leaf transcriptome dataset of coconut. Among the EST-SSRs, dinucleotide repeats were abundant (219,912; 69.04%) followed by trinucleotide (70,722; 22.2%) and tetra-nucleotide repeats (6281; 1.9%). Among the dinucleotide repeat motifs, the dominant repeat was AG/CT (35.87%), followed by AT/AT (18.59%), while the dominant trinucleotide repeat was AAG/CTT (4.59%). One hundred and twenty EST-SSR primer pairs were designed and utilized to amplify six DNA samples of coconut accessions. Fifty primers (41.7%) produced reproducible polymorphic fragments of expected sizes, from which a total of 10 primers were selected for the diversity assessment in 186 palms of 50 coconut accessions, comprising of 25 each of tall and dwarf accessions. A total of 137 alleles were detected with an average of 13.7 alleles per SSR locus. The number of alleles observed at each locus in the data set ranged from 7 to 22. All the loci showed 100% polymorphism with respect to the samples screened. The average observed heterozygosity was 0.46. The PIC values ranged from 0.79 (CnKGDEST129 and CnKGDEST100) to 0.91 (CnKGDEST117 and CnKGDEST122) with a mean value of 0.85, indicating the capacity of the EST-SSR markers to detect high levels of polymorphism. The cluster analysis revealed that accessions were generally clustered based on their relative similarity and irrespective of their geographic origins. The present study demonstrates the usefulness of transcriptome sequencing as a rapid and cost-effective methodology for the development of molecular markers. The EST-SSR markers generated through this study constitute useful and reliable tools for assessment of genetic diversity and marker-assisted selection in coconut.

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Funding

This research was funded by Indian Council of Agricultural Research (ICAR-CPCRI Institute Project Code No. 1000761030).

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Author notes

  1. S. Naganeeswaran

    Present address: Khalifa Center for Genetic Engineering and Biotechnology, United Arab Emirates University, Al-Ain, United Arab Emirates

  2. B. A. Jerard

    Present address: ICAR-Central Island Agricultural Research Institute, Port Blair, Andaman and Nicobar Islands, India

Authors and Affiliations

  1. ICAR-Central Plantation Crops Research Institute, Kasaragod, Kerala, India

    P. Preethi, Shafeeq Rahman, S. Naganeeswaran, A. A. Sabana, K. P. Gangaraj, B. A. Jerard, V. Niral & M. K. Rajesh

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Contributions

MKR designed the study; BAJ and VN provided the samples; PP, SR and SN performed the experiments; PP, AAS, KPG and MKR analyzed the data; PP wrote the paper; MKR, VN and BAJ checked the paper. The decision to submit the manuscript for publication was made by all the authors. All authors have read and approved the final manuscript.

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Correspondence to M. K. Rajesh.

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Preethi, P., Rahman, S., Naganeeswaran, S. et al. Development of EST-SSR markers for genetic diversity analysis in coconut (Cocos nucifera L.). Mol Biol Rep 47, 9385–9397 (2020). https://doi.org/10.1007/s11033-020-05981-8

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