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以欧洲葡萄‘佳丽酿’(Carinena)为材料,利用RT-PCR方法获得两个CIPK基因的全长cDNA序列,分别为VvCIPK13和VvCIPK14。VvCIPK13全长为1 501 bp,包括一个1 395 bp的ORF,编码465个氨基酸;VvCIPK14全长为1 616 bp,包括一个1 404 bp的ORF,编码468个氨基酸。生物信息学分析表明两个蛋白均含有两个保守结构域:丝氨酸/苏氨酸激酶结构域和NAF结构域,两个结构域之间有连接域,但VvCIPK13和VvCIPK14的同源性较低,仅有40.33%。利用荧光定量PCR技术研究VvCIPK13和VvCIPK14在葡萄不同组织、植物生长调节剂诱导和逆境胁迫下的表达模式,结果表明:VvCIPK13和VvCIPK14表达具有组织特异性,均在卷须中高表达;VvCIPK13对6-BA诱导有响应,而VvCIPK14对6-BA、ABA、IAA、SA、MeJA和GA_3等诱导均有不同程度的响应;VvCIPK13和VvCIPK14均响应高盐和干旱,但不响应低温,其中VvCIPK14的响应最为强烈。推测VvCIPK13和VvCIPK14在葡萄的非生物胁迫过程中发挥重要的作用。
Using the European grape Carinena as material, the full-length cDNA sequences of two CIPK genes were obtained by RT-PCR, which were VvCIPK13 and VvCIPK14, respectively. The full length of VvCIPK13 is 1 501 bp, including a 1 395 bp ORF encoding 465 amino acids. The full length of VvCIPK14 is 1 616 bp, including a 1 404 bp ORF encoding 468 amino acids. Bioinformatics analysis showed that both proteins contained two conserved domains: serine / threonine kinase domain and NAF domain, and there was a linkage domain between the two domains, but the homology of VvCIPK13 and VvCIPK14 was low, Only 40.33%. The expression patterns of VvCIPK13 and VvCIPK14 in different tissues of grape and plant growth regulators induced by stress and stress were studied by real-time fluorescence quantitative PCR. The results showed that the expression of VvCIPK13 and VvCIPK14 was tissue-specific, VvCIPK14 responded to 6-BA, ABA, IAA, SA, MeJA and GA_3, respectively. Both VvCIPK13 and VvCIPK14 responded to high salt and drought, but did not respond to low temperature. Among them, VvCIPK14 had the strongest response . It is speculated that VvCIPK13 and VvCIPK14 play an important role in the abiotic stress of grapes.