Plant Cell Reports

, Volume 27, Issue 11, pp 1741–1754

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
Genetic Transformation and Hybridization

DOI: 10.1007/s00299-008-0598-6

Cite this article as:
Konagaya, K., Ando, S., Kamachi, S. et al. Plant Cell Rep (2008) 27: 1741. doi:10.1007/s00299-008-0598-6

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.

Keywords

Male sterilityTapetumCysteine proteaseGene flowProgrammed cell death

Abbreviations

CaMV

Cauliflower mosaic virus

CKX

Cytokinin oxidase/dehydrogenase

GA

Gibberellin

GM

Genetically modified

GUS

β-Glucuronidase

ORF

Open reading frame

PCR

Polymerase chain reaction

RACE

Rapid amplification of cDNA ends

PCD

Programmed cell death

TAIL-PCR

Thermal asymmetric interlaced PCR

Supplementary material

299_2008_598_MOESM1_ESM.bmp (27.8 mb)
Supplementary figure S1 (BMP 28476 kb)
299_2008_598_MOESM2_ESM.bmp (61.8 mb)
Supplementary figure S2 (BMP 63245 kb)

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Ken-ichi Konagaya
    • 1
  • Sugihiro Ando
    • 1
  • Shinichiro Kamachi
    • 1
  • Mai Tsuda
    • 1
  • Yutaka Tabei
    • 1
  1. 1.Division of Plant ScienceNational Institute of Agrobiological SciencesIbarakiJapan