PLoS ONE (Jan 2023)

A pilot-scale comparison between single and double-digest RAD markers generated using GBS strategy in sesame (Sesamum indicum L.).

  • Pradeep Ruperao,
  • Prasad Bajaj,
  • Rajkumar Subramani,
  • Rashmi Yadav,
  • Vijaya Bhaskar Reddy Lachagari,
  • Sivarama Prasad Lekkala,
  • Abhishek Rathore,
  • Sunil Archak,
  • Ulavappa B Angadi,
  • Rakesh Singh,
  • Kuldeep Singh,
  • Sean Mayes,
  • Parimalan Rangan

DOI
https://doi.org/10.1371/journal.pone.0286599
Journal volume & issue
Vol. 18, no. 6
p. e0286599

Abstract

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To reduce the genome sequence representation, restriction site-associated DNA sequencing (RAD-seq) protocols is being widely used either with single-digest or double-digest methods. In this study, we genotyped the sesame population (48 sample size) in a pilot scale to compare single and double-digest RAD-seq (sd and ddRAD-seq) methods. We analysed the resulting short-read data generated from both protocols and assessed their performance impacting the downstream analysis using various parameters. The distinct k-mer count and gene presence absence variation (PAV) showed a significant difference between the sesame samples studied. Additionally, the variant calling from both datasets (sdRAD-seq and ddRAD-seq) exhibits a significant difference between them. The combined variants from both datasets helped in identifying the most diverse samples and possible sub-groups in the sesame population. The most diverse samples identified from each analysis (k-mer, gene PAV, SNP count, Heterozygosity, NJ and PCA) can possibly be representative samples holding major diversity of the small sesame population used in this study. The best possible strategies with suggested inputs for modifications to utilize the RAD-seq strategy efficiently on a large dataset containing thousands of samples to be subjected to molecular analysis like diversity, population structure and core development studies were discussed.