A TILLING Platform for Functional Genomics in Brachypodium distachyon

PLoS One. 2013 Jun 19;8(6):e65503. doi: 10.1371/journal.pone.0065503. Print 2013.

Abstract

The new model plant for temperate grasses, Brachypodium distachyon offers great potential as a tool for functional genomics. We have established a sodium azide-induced mutant collection and a TILLING platform, called "BRACHYTIL", for the inbred line Bd21-3. The TILLING collection consists of DNA isolated from 5530 different families. Phenotypes were reported and organized in a phenotypic tree that is freely available online. The tilling platform was validated by the isolation of mutants for seven genes belonging to multigene families of the lignin biosynthesis pathway. In particular, a large allelic series for BdCOMT6, a caffeic acid O-methyl transferase was identified. Some mutants show lower lignin content when compared to wild-type plants as well as a typical decrease of syringyl units, a hallmark of COMT-deficient plants. The mutation rate was estimated at one mutation per 396 kb, or an average of 680 mutations per line. The collection was also used to assess the Genetically Effective Cell Number that was shown to be at least equal to 4 cells in Brachypodium distachyon. The mutant population and the TILLING platform should greatly facilitate functional genomics approaches in this model organism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosynthetic Pathways
  • Brachypodium / genetics
  • Brachypodium / growth & development*
  • Brachypodium / metabolism
  • Genome, Plant
  • Genomics / methods*
  • Lignin / metabolism
  • Models, Molecular
  • Mutation*
  • Phenotype
  • Phylogeny
  • Plant Breeding
  • Plant Proteins / chemistry
  • Plant Proteins / genetics*
  • Sequence Analysis, DNA

Substances

  • Plant Proteins
  • Lignin

Grant support

This work was funded by the European FP7 project n° 211982 “RENEWALL” and the Agence Nationale de la Recherche (ANR) projects ANR-08-KBBE-004 “CELLWALL” and ANR-10-BLAN-1528 “PHENOWALL”. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.