葡萄VvSCL9基因和启动子的克隆及初步研究  

Isolation and primary function study of VvSCL9 gene and its promoter

作　　者：陈立勇[1] 陈丽华[1] 柴丽娟[1] 陈尚武[2] 马会勤[1] 

机构地区：[1]中国农业大学农学与生物技术学院,北京100193 [2]中国农业大学食品科学与营养工程学院,北京100193

出　　处：《中国农业大学学报》2015年第2期101-107,共7页Journal of China Agricultural University

基　　金：国家自然科学基金项目(31171939)

摘　　要：以欧亚种无核葡萄品种‘无核白鸡心’(Vitisvinifera L.cv.Centennial Seedless)为材料,通过半定量RTPCR比较了VvSCL9基因在葡萄不同组织器官中的表达,利用转录组分析探索了葡萄果实中VvSCL9对外源赤霉素(GA3)处理的响应。通过全基因序列和启动子克隆、启动子生物信息学分析及转基因验证进一步研究了VvSCL9的表达特征及对不同胁迫的响应。结果表明,VvSCL9在葡萄梢尖、幼叶、成叶、老叶和花序等组织器官中均有表达,参与葡萄果实第一快速生长期的发育,但对外源GA3处理无显著响应。对克隆得到的VvSCL9启动子进行生物信息学分析,发现多个激素和逆境胁迫响应的作用元件。以GA3处理超表达VvSCL9的转基因烟草种子,转基因烟草种子的萌发率在前期低于对照。转基因烟草组培苗对100mmol/L NaCl的耐受性高于对照。研究表明,VvSCL9基因参与葡萄的营养生长和生殖生长,能够提高植物耐盐性,在一定程度上影响植物外源GA3的敏感性,但可能不是外源赤霉素(GA3)介导的无核葡萄果实膨大过程的主要调控元件。SCARECROW-Like（SCL）genes are the members of GRAS gene family.They involve in many processes including plant development,hormone and stress response.In the present study,‘Centennial Seedless＇（Vitisvinifera L.）variety was used to detect the expression of VvSCL9 in different tissues and organsby a semi-quantitative RT-PCR.The precise transcription abundance in berries after exogenous GA3 treatment was revealed by transcriptomic data analysis.The expression character and function of VvSCL9 were further studied by full sequence and promoter cloning,bioinformatic analysis and heterogeneous expression.The semi-quantitative RT-PCR detected VvSCL9 expression in shoot tips,young leaves,full-expanded leaves,old leaves and inflorescence.Berry transcriptomic data revealed that VvSCL9 was expressed in stage I berry.But no differential expression was found on 1,3 and 7dafter exogenous GA3 application.Bioinformatics analysis found that VvSCL9 promoter had multi-acting elements involved in response to plant hormones and stress resistance.Transgenic tobacco seeds over-expressing VvSCL9 demonstrated lower germination rate than the controls in the early days under GA3 treatment.Transgenic tobacco plants over-expressing VvSCL9 improved the plant tolerance to 100 mmol/L NaCl.Our results indicated that VvSCL9 participated invegetative and reproductive growth of grapevine,it involved in plant salt tolerance and the sensitivity to GA3.However,VvSCL9 might not serve as a main regulatory element in exogenous gibberellins（GA3）induced seedless grape berry enlargement.

关 键 词：葡萄 VvSCL9基因 启动子 GA3处理 盐胁迫 

分 类 号：S663.1[农业科学—果树学]