Rapid identification of causal mutations in tomato EMS populations via mapping-by-sequencing. - INRAE - Institut national de recherche pour l’agriculture, l’alimentation et l’environnement Access content directly
Journal Articles Nature Protocols Year : 2016

Rapid identification of causal mutations in tomato EMS populations via mapping-by-sequencing.

Virginie Garcia
Cécile Bres
Daniel Just
Lucie Fernandez
Jean-Philippe Mauxion
  • Function : Author
  • PersonId : 1137443
  • IdHAL : jp-mauxion
Koh Aoki
  • Function : Author
Christophe Rothan

Abstract

The tomato is the model species of choice for fleshy fruit development and for the Solanaceae family. Ethyl methanesulfonate (EMS) mutants of tomato have already proven their utility for analysis of gene function in plants, leading to improved breeding stocks and superior tomato varieties. However, until recently, the identification of causal mutations that underlie particular phenotypes has been a very lengthy task that many laboratories could not afford because of spatial and technical limitations. Here, we describe a simple protocol for identifying causal mutations in tomato using a mapping-by-sequencing strategy. Plants displaying phenotypes of interest are first isolated by screening an EMS mutant collection generated in the miniature cultivar Micro-Tom. A recombinant F2 population is then produced by crossing the mutant with a wild-type (WT; non-mutagenized) genotype, and F2 segregants displaying the same phenotype are subsequently pooled. Finally, whole-genome sequencing and analysis of allele distributions in the pools allow for the identification of the causal mutation. The whole process, from the isolation of the tomato mutant to the identification of the causal mutation, takes 6-12 months. This strategy overcomes many previous limitations, is simple to use and can be applied in most laboratories with limited facilities for plant culture and genotyping.
No file

Dates and versions

hal-02638680 , version 1 (28-05-2020)

Identifiers

Cite

Virginie Garcia, Cécile Bres, Daniel Just, Lucie Fernandez, Virginie Wong Jun Tai, et al.. Rapid identification of causal mutations in tomato EMS populations via mapping-by-sequencing.. Nature Protocols, 2016, 11 (12), pp.2401-2418. ⟨10.1038/nprot.2016.143⟩. ⟨hal-02638680⟩
45 View
0 Download

Altmetric

Share

Gmail Mastodon Facebook X LinkedIn More