Over-expression of PR-10a leads to increased salt and osmotic tolerance in potato cell cultures.
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.
El-Banna
Antar
A
German Collection of Microorganisms and Cell Cultures GmbH, Inhoffenstr. 7b, D-38124 Braunschweig, Germany.
Hajirezaei
Mohammad-Reza
MR
Wissing
Joseph
J
Ali
Zahid
Z
Vaas
Lea
L
Heine-Dobbernack
Elke
E
Jacobsen
Hans-Jörg
HJ
Schumacher
Heinz Martin
HM
Kiesecker
Heiko
H
D015139:Blotting, Southern; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005978:Glutathione / Q000378:metabolism; D009687:Nuclear Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D009997:Osmotic Pressure / Q000502:physiology*; D010940:Plant Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D011392:Proline / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D018749:RNA, Plant / Q000032:analysis / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012965:Sodium Chloride / Q000494:pharmacology; D011198:Solanum tuberosum / Q000235:genetics / Q000378:metabolism*; D013012:Sorbitol / Q000378:metabolism; D013312:Stress, Physiological / Q000502:physiology*; D014170:Transformation, Genetic
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C416516:PBF-2 protein, Solanum tuberosum; D010940:Plant Proteins; D012333:RNA, Messenger; D018749:RNA, Plant; D012965:Sodium Chloride; D013012:Sorbitol; D011392:Proline; D005978:Glutathione