Over-expression of PR-10a leads to increased salt and osmotic tolerance in potato cell cultures

J Biotechnol. 2010 Nov;150(3):277-87. doi: 10.1016/j.jbiotec.2010.09.934. Epub 2010 Sep 17.

Abstract

The PR-10a protein (formerly STH-2) is known to be induced by biotic stress in potato. The present study demonstrates that transgenic suspension cells of the potato cultivar Desiree over-expressing the PR-10a protein exhibit significantly increased salt and osmotic tolerance compared to the respective wild type cells. A comparison of the proteome pattern of Solanum tuberosum suspension cultures cv. Desiree before and after the treatment with NaCl or sorbitol under equiosmolar conditions (740mOs/kg) revealed the pathogenesis related protein PR-10a to be one of the predominant differentially expressed proteins in potato cell cultures. The pr-10a mRNA was confirmed to be present by RT-PCR from salt challenged suspension cells and was transcribed into cDNA. For PR-10a over-expression Agrobacterium tumefaciens mediated transformation of the potato cells and a dicistronic vector harboring the cDNA of the pr-10a gene linked to a luciferase gene by an IRES (Internal Ribosome Binding Site) was used. The IRES mediated translation leads to co-expression of PR-10a and luciferase in a fixed ratio. By non-invasive luciferase assay homologous PR-10a over-expressing callus was identified after selection on phosphinothricin supplemented medium. This callus was used for the setup of a transgenic suspension culture. Along with increased salt and osmotic tolerance the transformed culture showed changed proline and glutathione levels under abiotic stress conditions in comparison to the wild type.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Southern
  • Cells, Cultured
  • DNA-Binding Proteins / analysis
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Electrophoresis, Gel, Two-Dimensional
  • Glutathione / metabolism
  • Nuclear Proteins / analysis
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Osmotic Pressure / physiology*
  • Plant Proteins / analysis
  • Plant Proteins / genetics
  • Plant Proteins / metabolism*
  • Proline / metabolism
  • RNA, Messenger / analysis
  • RNA, Messenger / metabolism
  • RNA, Plant / analysis
  • RNA, Plant / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sodium Chloride / pharmacology
  • Solanum tuberosum / genetics
  • Solanum tuberosum / metabolism*
  • Sorbitol / metabolism
  • Stress, Physiological / physiology*
  • Transformation, Genetic

Substances

  • DNA-Binding Proteins
  • Nuclear Proteins
  • PBF-2 protein, Solanum tuberosum
  • Plant Proteins
  • RNA, Messenger
  • RNA, Plant
  • Sodium Chloride
  • Sorbitol
  • Proline
  • Glutathione