Gene editing a constitutively active osrac1 by homologous recombination-based gene targeting induces immune responses in rice

Thu Thi Dang, Zenpei Shimatani, Yoji Kawano, Rie Terada, Ko Shimamoto

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

OsRac1 is a member of the plant small GTPase Rac/Rop family and plays a key role in rice immunity. The constitutively active (CA) G19V mutation of OsRac1 was previously shown to induce reactive oxygen species production, phytoalexin synthesis and defense gene activation, leading to resistance to rice blast infection. To study further the effect of the G19V mutation in disease resistance, we introduced a single base substitution by gene targeting and removed the selectable marker using Cre-loxP site-specific recombination. The CA-OsRac1 gene generated by gene targeting was termed CA-gOsRac1. The G19V mutation was transferred from a targeting vector to the OsRac1 locus and stably transmitted to the next generation. In the leaf blade of homozygous CA-gOsRac1 plants, mutant transcript levels were much lower than in those of wild-type plants. In contrast, mutant transcripts in roots, leaf sheaths and panicles were more abundant than those in leaf blades. However, upon chitin treatment, the expression of defense-related genes PAL1 and PBZ1 in the cell culture was greater in the mutants compared with wild-type plants. Furthermore, induction of hypersensitive response (HR)-like cell death was observed in the leaf sheaths of mutant plants infected with a compatible race of rice blast fungus. In the CA-gOsRac1 plants, a number of genes previously shown to be induced by Magnaporthe oryzae and Xanthomonas oryzae pv. oryzae (Xoo) infection were induced in the leaf sheath without pathogen infection. These results suggest that gene targeting will provide mutations useful for gene function studies and crop improvement.

Original languageEnglish
Pages (from-to)2058-2070
Number of pages13
JournalPlant and Cell Physiology
Volume54
Issue number12
DOIs
Publication statusPublished - Dec 2013
Externally publishedYes

Keywords

  • Cre-loxP site-specific recombination
  • Gene targeting
  • Homologous recombination
  • Immune responses
  • Marker-free transgenic rice
  • OsRac1

ASJC Scopus subject areas

  • Physiology
  • Plant Science
  • Cell Biology

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