Expression of a pathogen-induced cysteine protease (AdCP) in tapetum results in male sterility in transgenic tobacco

Funct Integr Genomics. 2014 Jun;14(2):307-17. doi: 10.1007/s10142-014-0367-2. Epub 2014 Mar 11.

Abstract

Usable male sterility systems have immense potential in developing hybrid varieties in crop plants, which can also be used as a biological safety containment to prevent horizontal transgene flow. Barnase-Barstar system developed earlier was the first approach to engineer male sterility in plants. In an analogous situation, we have evolved a system of inducing pollen abortion and male sterility in transgenic tobacco by expressing a plant gene coding for a protein with known developmental function in contrast to the Barnase-Barstar system, which deploys genes of prokaryotic origin, i.e., from Bacillus amyloliquefaciens. We have used a plant pathogen-induced gene, cysteine protease for inducing male sterility. This gene was identified in the wild peanut, Arachis diogoi differentially expressed when it was challenged with the late leaf spot pathogen, Phaeoisariopsis personata. Arachis diogoi cysteine protease (AdCP) was expressed under the strong tapetum-specific promoter (TA29) and tobacco transformants were generated. Morphological and histological analysis of AdCP transgenic plants showed ablated tapetum and complete pollen abortion in three transgenic lines. Furthermore, transcript analysis displayed the expression of cysteine protease in these male sterile lines and the expression of the protein was identified in western blot analysis using its polyclonal antibody raised in the rabbit system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Arachis / genetics
  • Arachis / microbiology
  • Ascomycota / pathogenicity
  • Ascomycota / physiology
  • Cysteine Proteases / genetics*
  • Cysteine Proteases / metabolism
  • Gene Expression Regulation, Plant*
  • Molecular Sequence Data
  • Nicotiana / enzymology
  • Nicotiana / genetics*
  • Plant Infertility / genetics*
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism
  • Plants, Genetically Modified / enzymology
  • Plants, Genetically Modified / genetics*
  • Pollen
  • Promoter Regions, Genetic
  • Sequence Alignment
  • Transgenes

Substances

  • Plant Proteins
  • Cysteine Proteases