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磷脂酰肌醇转运蛋白Sec14是最先在酿酒酵母(Saccharomyces cerevisiae)中发现的磷脂酰肌醇转运蛋白,含有一个典型的SEC14保守结构域。Sec14p磷脂酰肌醇转运蛋白广泛存在于真核生物细胞中,并参与膜运输途径、质膜发育、脂肪酸代谢、根毛的生长等生命活动,但未见此蛋白在作物逆境胁迫的有关报道。为研究Sec14p基因在小麦(Triticum aestivum)发育及逆境胁迫中的功能,本研究采用电子克隆方法从小麦京花9号中得到一个Sec14p基因,命名为Ta SEC14p-5(Gen Bank登录号:KU639968)。氨基酸序列分析表明,该基因ORF全长为984 bp,编码327个氨基酸,相对分子质量为37.1 k D,理论等电点为7.70。在植物中该蛋白有典型的SEC14类基因家族保守域SEC14和一个CRAL_TRIO_N结构域。进化和聚类分析表明,小麦Ta SEC14p-5基因与粗山羊草(Aegilops tauschii)磷脂酰肌醇转运蛋白Ae SEC14p基因和水稻(Oryza sativa)磷脂酰肌醇转运蛋白Os02g0200000基因亲缘关系较近,蛋白相似度分别为96.45%和88.38%。采用q RT-PCR技术,对小麦孕穗期不同组织和不同非生物胁迫幼苗进行表达分析。发现该基因在根、茎、叶、颖壳、雄蕊和种子中均有表达,且在茎中表达量较高,雄蕊次之,根最低;该基因受高盐的强烈诱导,同时也受到脱落酸(abscisic acid,ABA)(200μmol/L)、干旱(PEG6000)和低温(4℃)不同程度的胁迫诱导。Ta SEC14p-5在Na Cl、ABA、PEG和4℃四种不同胁迫处理下,总体趋势为先上升后下降。结果推测,Ta SEC14p-5基因可能参与了Na Cl、ABA、PEG6000和4℃等不同程度的非生物胁迫处理,为进一步改良小麦抗逆性等分子机理研究提供了新的依据。
Phosphatidylinositol transporter Sec14 is the first phosphatidylinositol transporter found in Saccharomyces cerevisiae and contains a typical SEC14 conserved domain. Sec14p phosphatidylinositol transporter is widely present in eukaryotic cells and participates in the life activities such as membrane transport, plasma membrane development, fatty acid metabolism and root hair growth, but no reports of this protein in crop stress are reported. In order to study the function of Sec14p gene in development and stress of Triticum aestivum, a Sec14p gene named Ta SEC14p-5 (GenBank accession number: KU639968) was obtained from wheat Jinghua 9 by using electronic cloning method. . Amino acid sequence analysis showed that the full-length ORF of this gene was 984 bp, encoding 327 amino acids with a relative molecular mass of 37.1 kD and a theoretical isoelectric point of 7.70. In plants, the protein has a typical SEC14 gene family conserved domain SEC14 and a CRAL_TRIO_N domain. Phylogenetic analysis and cluster analysis showed that the Ta SEC14p-5 gene in wheat is closely related to the Ae SEC24p gene of Aegilops tauschii and the Os02g0200000 gene of rice Oryza sativa phosphatidylinositol transporter, Similarities were 96.45% and 88.38% respectively. Q RT-PCR was used to analyze the expression of different tissues and different abiotic stress seedlings in booting stage of wheat. The gene was found to be expressed in roots, stems, leaves, glume, stamens and seeds, with higher expression in stalks, followed by stamens and lowest in roots. The gene was strongly induced by high salt and was also shed Abscisic acid (ABA) (200μmol / L), drought (PEG6000) and low temperature (4 ℃) were induced by different degrees of stress. Ta SEC14p-5 in Na Cl, ABA, PEG and 4 ℃ under four different stress treatment, the overall trend for the first rise and then decline. The results suggest that Ta SEC14p-5 may be involved in abiotic stresses such as Na Cl, ABA, PEG6000 and 4 ℃, which provide a new basis for further molecular mechanism studies such as stress tolerance.