Expression of Acyl‐lipid Δ12‐desaturase Gene in Prokaryotic and Eukaryotic Cells and Its Effect on Cold Stress Tolerance of Potato

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.

RNA-seq based identification and mutant validation of gene targets related to ethanol resistance in cyanobacterial Synechocystis sp. PCC 6803

“…RNA-seq transcriptomics analysis showed that slr1350 encoding acyl-lipid desaturase was up-regulated at 72 h. In a previous study, the acyl-lipid desaturase ( desA ) gene from Synechocystis sp. PCC6803 was expressed in prokaryotic ( E. coli ) and eukaryotic ( Solanum tuberosum ) cells, which led to an enhanced cold tolerance due to increased unsaturated fatty acid concentration in their lipids [32]. Several genes encoding cell envelope proteins were found induced by ethanol exposure (Table 3).…”

A comparative study of fatty acid profile and formation of biofilm inGeobacillus gargensisexposed to variable abiotic stress

“…Geobacillus gargensis showed an increased synthesis of BCFAs and SCSFAs at 60-65°C, indicating an activation of branchedchain alpha-keto acid reductase (BCKR). At lower culture temperatures, desaturases are upregulated with an increase in unsaturated FAs (Amiri et al 2010). These changes reflect on the dynamic and physiological responses of G. gargensis subjected to variable temperature regimes.…”

Proteomic Analysis and qRT-PCR Verification of Temperature Response to Arthrospira (Spirulina) platensis

“…However as a response to high temperature, the denatured proteins increase in vivo , and Hsp60 binds to the denatured protein to maintain their soluble state. In the presence of Mg 2+ -ATP, Hsp60 can change folding proteins to refold into the active conformation, or degrade the denatured protein [35]. For example, Hsp60 can refold the Rubisco enzyme of cyanobacteria and malate dehydrogenase of mitochondrion in vitro [36].…”