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| Cloning and Expression Analysis of TaSEC14p-5 Gene from Wheat (Triticum aestivum) |
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Abstract Sec14p, found in yeast (Saccharomyces cerevisiae) and containing a typical conserved domain SEC14, is as research object possessing the same function with phosphatidylinositol transfer protein. Sec14p phosphatidylinositol transfer protein exists widely in eukaryotic cells and may be involved in a variety of biological activities, such as membrane trafficking, plasma membrane development, fatty acid metabolism, yeast microspore formation and the growth of plant root hair. However, it's relationship with plant environmental stresses have not been reported. In order to study the function and mechanism of Sec14 superfamily in wheat (Triticum aestivum) and further enrich the database of wheat genome, in silico cloning technique was used to clone TaSEC14p-5 gene (GenBank No. KU639968) from the genome of wheat cultivar Jinghua 9. The analysis of nucleotide sequence indicated that TaSEC14p-5 gene sequence contained a 984 bp intact ORF, which encoded a peptide of 327 amino acids. The predicted molecular weight and isoelctric point of TaSEC14p-5 were 37.1 kD and 7.70, respectively. The amino acids analysis indicated that the predicted protein sequence contained a typical SEC14 superfamily domain and a CRAL_TRIO_N binding domain in plant. Evolution and clustering analysis revealed that TaSEC14p-5 protein shared 96.45% and 88.38% sequence similarities with AeSEC14p (Aegilops tauschii) and Os02g0200000 (Oryza sativa) phosphatidylinositol transfer protein. The expression pattern analysis, carried out by qRT-PCR, indicated that TaSEC14p-5 was constitutively expressed in various tissues of wheat at the booting stage, with higher level in stem, and then in stamen, but less in root. Meanwhile, it was induced by abscisic (ABA) (200 μmol/L), drought (PEG6000), high-salt (250 mmol/L NaCl) and cold (4 ℃) stresses, but under high salinity it was strongly up-regulated. This research aimed at identificating a novel wheat Sec14p phosphatidylinositol transfer protein gene (TaSEC14p-5) relative transcript level in the above mentioned 4 stress treatments. The conclusion was that the novel TaSEC14p-5 gene was an excellent candidate gene, because it was involved in a variety of signal transduction pathway adverse treatment. Otherwise, the gene of interest provides a new research direction for improving resistance and other molecular mechanism in wheat.
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Received: 01 February 2016
Published: 01 July 2016
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