Efficient production of genetically engineered, male-sterile Arabidopsis thaliana using anther-specific promoters and genes derived from Brassica oleracea and B. rapa

Ken Ichi Konagaya, Sugihiro Ando, Shinichiro Kamachi, Mai Tsuda, Yutaka Tabei

Research output: Contribution to journalArticlepeer-review

34 Citations (Scopus)

Abstract

Prevention of transgene flow from genetically modified crops to food crops and wild relatives is of concern in agricultural biotechnology. We used genes derived from food crops to produce complete male sterility as a strategy for gene confinement as well as to reduce the food purity concerns of consumers. Anther-specific promoters (A3, A6, A9, MS2, and MS5) were isolated from Brassica oleracea and B. rapa and fused to the β-glucuronidase (GUS) reporter gene and candidate genes for male sterility, including the cysteine proteases BoCysP1 and BoCP3, and negative regulatory components of phytohormonal responses involved in male development. These constructs were then introduced into Arabidopsis thaliana. GUS analyses revealed that A3, A6, and A9 had tapetum-specific promoter activity from the anther meiocyte stage. Male sterility was confirmed in tested constructs with protease or gibberellin insensitive (gai) genes. In particular, constructs with BoCysP1 driven by the A3 or A9 promoter most efficiently produced plants with complete male sterility. The tapetum and middle layer cells of anthers expressing BoCysP1 were swollen and excessively vacuolated when observed in transverse section. This suggests that the ectopic expression of cysteine protease in the meiocyte stage may inhibit programmed cell death. The gai gene also induced male sterility, although at a low frequency. This is the first report to show that plant cysteine proteases and gai from food crops are available as a novel tool for the development of genetically engineered male-sterile plants.

Original languageEnglish
Pages (from-to)1741-1754
Number of pages14
JournalPlant Cell Reports
Volume27
Issue number11
DOIs
Publication statusPublished - 2008 Nov
Externally publishedYes

Keywords

  • Cysteine protease
  • Gene flow
  • Male sterility
  • Programmed cell death
  • Tapetum

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science

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