Expression of acyl-lipid Delta12-desaturase gene in prokaryotic and eukaryotic cells and its effect on cold stress tolerance of potato

J Integr Plant Biol. 2010 Mar;52(3):289-97. doi: 10.1111/j.1744-7909.2010.00890.x.

Abstract

We report the expression profile of acyl-lipid Delta12-desaturase (desA) gene from Synechocystis sp. PCC6803 and its effect on cell membrane lipid composition and cold tolerance in prokaryotic (Escherichia coli) and eukaryotic (Solanum tuberosum) cells. For this purpose, a hybrid of desA and reporter gene encoding thermostable lichenase (licBM3) was constructed and used to transform these cells. The expression of this hybrid gene was measured using qualitative (Petri dish test, electrophoregram and zymogram) and quantitative methods (spectrometry and gas liquid chromatography assays). The maximum level of linoleic acid in the bacterial cells containing hybrid gene was 1.9% of total fatty acids. Cold stress tolerance assays using plant damage index and growth parameters showed that cold tolerance was enhanced in primary transgenic lines because of increased unsaturated fatty acid concentration in their lipids. The greatest content of 18:2 and 18:3 fatty acids in primary transgenic plants was observed for lines 2 (73%) and 3 (41%). Finally, our results showed that desaturase could enhance tolerance to cold stress in potato, and desaturase and lichenase retain their functionality in the structure of the hybrid protein where the enzymatic activity of target gene product was higher than in the case of reporter lichenase gene absence in the construction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptation, Physiological* / genetics
  • Cold Temperature*
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Stability
  • Escherichia coli / metabolism
  • Eukaryotic Cells / metabolism*
  • Fatty Acid Desaturases / genetics*
  • Gene Expression Regulation, Plant
  • Glycoside Hydrolases / genetics
  • Glycoside Hydrolases / metabolism
  • Linoleic Acids
  • Plant Leaves / metabolism
  • Plants, Genetically Modified
  • Prokaryotic Cells / metabolism*
  • Solanum tuberosum / genetics
  • Solanum tuberosum / growth & development
  • Solanum tuberosum / physiology*
  • Stress, Physiological / genetics
  • Synechocystis / enzymology*
  • Synechocystis / genetics
  • Time Factors
  • Transformation, Genetic

Substances

  • Linoleic Acids
  • Fatty Acid Desaturases
  • Glycoside Hydrolases
  • licheninase