Identification of Solanum Immune Receptors by Bulked Segregant RNA-Seq and High-Throughput Recombinant Screening

Yerisf Torres Ascurra, Xiao Lin, Pieter J. Wolters, Vivianne G.A.A. Vleeshouwers*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

2 Citations (Scopus)

Abstract

The identification, understanding, and deployment of immune receptors are crucial to achieve high-level and durable resistance for crops against pathogens. In potato, many R genes have been identified using map-based cloning strategies. However, this is a challenging and laborious task that involves the development of a high number of molecular markers for the initial mapping, and the screening of thousands of plants for fine mapping. Bulked segregant RNA-Seq (BSR-Seq) has proven to be an efficient technique for the mapping of resistance genes. The RNA from two bulks of plants with contrasting phenotypes is sequenced and analyzed to identify single-nucleotide polymorphism (SNPs) markers linked to the target gene. Subsequently, the SNP markers that are identified can be used to delimit the mapping interval. Additionally, we designed an in vitro recombinant screening strategy that is advantageous for analyzing a large number of plants, in terms of time, space, and cost. Tips and detailed protocols, including BSR-Seq, bioinformatic analysis, and recombinant screening, are provided in this chapter.

Original languageEnglish
Title of host publicationSolanum tuberosum
Subtitle of host publicationMethods and Protocols
EditorsD. Dobnik, K. Gruden, Z. Ramsak, A. Coll
Place of PublicationNew York
PublisherHumana Press
Pages315-330
Number of pages16
ISBN (Electronic)9781071616093
ISBN (Print)9781071616086
DOIs
Publication statusPublished - 27 Aug 2021

Publication series

NameMethods in Molecular Biology
Volume2354
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Bulked segregant analysis (BSA)
  • Effectoromics
  • Effectors
  • Genetic mapping
  • Mapping
  • Nucleotide-binding site leucine-rich repeat (NLR)
  • Pattern recognition receptors (PRR)
  • Receptor-like kinases (RLK)
  • Receptor-like proteins (RLP)
  • Resistance genes (R genes)

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