山葡萄VaCBL6参与非生物胁迫和ABA途径的响应

VaCBL6 from Vitis amurensis Involved in Abiotic Stress Response and ABA-mediated Pathway

对山葡萄(Vitisamurensis)‘左山-1’中的1个类钙调素磷酸酶B亚基蛋白基因VaCBL6进行克隆及功能分析。VaCBL6位于第4条染色体,开放阅读框为777 bp,编码258个氨基酸;结构域分析表明,该蛋白包含4个EF-hand与1个跨膜结构域;蛋白进化树分析表明,VaCBL6与欧洲葡萄VvCBL9和VvCBL6相似性较高,分别为98.84%、94.96%。VaCBL6编码蛋白定位于细胞核与细胞膜。VaCBL6在根与卷须中有较高的表达,且其表达受低温、干旱、NaCl和ABA的诱导。基于酵母体系的转录激活分析表明,VaCBL6无转录激活活性。100~125μmol·L^(-1) NaCl和0.25~1.25μmol·L^(-1) ABA处理均抑制了VaCBL6转基因拟南芥种子的萌发,且转化拟南芥幼苗对NaCl与ABA的敏感性提高。以上结果说明VaCBL6在植物的逆境胁迫应答中具有重要作用。

A calcineurin B-like phosphatase subunit protein gene VaCBL6 from Vitis amurensis’Zuoshan-1’was cloned and functionally analyzed.VaCBL6 is located on chromosome 4 and has an open reading frame of 777 bp,encoding 258 aa;protein domain analysis shows that the protein contains four EF-hands and one transmembrane domain.Phylogenetic analysis using amino acid sequence showed that high similarities between VaCBL6 and Vv CBL9,VaCBL6 and Vv CBL6 are 98.84%and 94.96%,respectively.VaCBL6 is located in the nucleus and cell membrane,and is highly expressed in roots and tendrils,and its expression is induced by low temperature,drought,NaCl and ABA.Analysis of transcriptional activation based on the yeast system displayed that VaCBL6 had no transcriptional activation activity.Experiments on the growth stage of Arabidopsis seedlings overexpressing VaCBL6 with different treatments found that 100-125μmol·L^(-1) NaCl and 0.25-1.25μmol·L^(-1) ABA treatments inhibited the germination of VaCBL6 transgenic Arabidopsis seeds,and the transgenic Arabidopsis seedlings increased the sensitivity to NaCl and ABA.The results indicated that VaCBL6 plays an important role in plant stress response.