TILLING in barley

Matthias Jost, Miriam Szurman-Zubrzycka, Katarzyna Gajek, Iwona Szarejko, Nils Stein

Research output: Chapter in Book/Conference paperChapterpeer-review

5 Citations (Scopus)

Abstract

TILLING (Targeting Induced Local Lesions IN Genomes), a popular reverse genetics approach in barley research, combines plant mutagenesis with efficient mutation detection for studying biological function of a specific gene. The high mutation frequency within a TILLING population principally enables the identification of induced variations in (almost) all genes of a given species (more precisely a given genotype of a species) of interest, which can be tested for their functional impact on morphological and/or physiological characteristics of the plant. Several TILLING populations induced by chemical mutagenesis were established for barley (Talame et al., Plant Biotechnol J 6:477–485, 2008; Gottwald et al., BMC Res Notes 2:258, 2009; Caldwell et al. Plant J 40:143–150, 2004) and showed the possibility for adapting protocols to develop further populations. This chapter describes a chemical mutagenesis protocol for barley seeds and two independent procedures for efficient single nucleotide polymorphism (SNP) detection in a large number of mutagenized plants either by slab-gel- or capillary gel-based electrophoreses on the LI-COR 4300 DNA Analyzer and the AdvanCE FS96 instruments, respectively.

Original languageEnglish
Title of host publicationBarley
Subtitle of host publicationMethods and Protocols
EditorsWendy A. Harwood
Place of PublicationNew York
PublisherHumana Press
Pages73-94
Number of pages22
ISBN (Electronic)978-1-4939-8944-7
ISBN (Print)978-1-4939-8942-3
DOIs
Publication statusPublished - 2019
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1900
ISSN (Print)1064-3745

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