Development of user-friendly markers for disease resistance to black root rot of tobacco through genotyping by sequencing

Qiulin Qin, Yuting Li, Na Ding, Dandan Li, Natalia Martinez, Robert Miller, David Zaitlin, Shengming Yang

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Black root rot (BRR), a disease caused by the hemibiotrophic fungus Thielaviopsis basicola, seriously compromises yield and leaf quality in tobacco (Nicotiana tabacum). Full resistance to black root rot, conferred by the resistance to BRR 1 (RBRR1) locus from Nicotiana debneyi Domin, was transferred to a burley tobacco cultivar through interspecific hybridization. Some undesirable traits potentially caused by linkage drag restrict wider application of RBRR1 in flue-cured tobacco. Therefore, user-friendly molecular markers are needed to assist selection for resistance to black root rot and to break the unfavorable linkage. Genotyping by sequencing (GBS) is a rapid and robust approach for reduced representation sequencing of multiplexed genomic DNA samples that combines genome-wide molecular marker discovery with genotyping. In the present study, we used GBS to identify single-nucleotide polymorphisms (SNPs) linked to the RBRR1 locus, and PCR-based assays for detection of these SNPs were also developed. Sequence analysis of the SNP markers suggested that RBRR1 is located on chromosome 17, providing a basis for map-based cloning of this valuable gene. Co-dominant CAPS markers that co-segregate with the disease-resistant phenotype offer user-friendly tools for tobacco breeding and variety improvement. Furthermore, tested with diverse N. tabacum germplasm, SS192650 displayed 100% selection accuracy for resistance to BRR, suggesting that this marker can be used in diverse tobacco populations.

Original languageEnglish
Article number76
JournalMolecular Breeding
Volume38
Issue number6
DOIs
StatePublished - Jun 1 2018

Bibliographical note

Funding Information:
Author contributions Conceived and designed the experiments: DL, RM, DZ, and SY. Performed the experiments: QQ, YL, ND, DL, and NM. Analyzed the data: QQ, YL, and SY. Wrote the first draft: QQ and SY.Funding informationThis research was supported by British American Tobacco (BAT), Council for Burley Tobacco, and Kentucky Tobacco Research & Development Center (KTRDC).

Publisher Copyright:
© 2018, Springer Science+Business Media B.V., part of Springer Nature.

Keywords

  • Black root rot
  • GBS
  • Nicotiana tabacum
  • SNPs
  • User-friendly marker

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Agronomy and Crop Science
  • Genetics
  • Plant Science

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