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

• Published in: Journal of biotechnology 2010-11, Vol.150 (3), p.277-287
• Main Authors: El-Banna, Antar, Hajirezaei, Mohammad-Reza, Wissing, Joseph, Ali, Zahid, Vaas, Lea, Heine-Dobbernack, Elke, Jacobsen, Hans-Jörg, Schumacher, Heinz Martin, Kiesecker, Heiko
• Format: Article
• Language: English
• Subjects: Agrobacterium tumefaciens; Biological and medical sciences; Biotechnology; Blotting, Southern; Cells, Cultured; DNA-Binding Proteins - analysis; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Electrophoresis, Gel, Two-Dimensional; Fundamental and applied biological sciences. Psychology; Glutathione - metabolism; Nuclear Proteins - analysis; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; Osmotic Pressure - physiology; Osmotic stress; Plant Proteins - analysis; Plant Proteins - genetics; Plant Proteins - metabolism; Potato; PR-10a; Proline - metabolism; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - analysis; RNA, Messenger - metabolism; RNA, Plant - analysis; RNA, Plant - metabolism; Salt stress; Sodium Chloride - pharmacology; Solanum tuberosum; Solanum tuberosum - genetics; Solanum tuberosum - metabolism; Sorbitol - metabolism; Stress, Physiological - physiology; Suspension culture; Transformation, Genetic
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2010.09.934

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 (740 mOs/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.