葡萄SnRK2家族基因的鉴定与表达分析  

Identification and Expression Profile of the SnRK2 Family Genes in Grapevine

作　　者：马宗桓[1] 毛娟[1] 李文芳[1] 杨世茂[1] 吴金红[1] 陈佰鸿[1] MAZong-huan MAOJuan LIWen-fang YANGShi-mao WUJin-hong and CHENBai-hong(College of Horticulture, Gansu Agricultural University, Lanzhou 730070, Chin)

机构地区：[1]甘肃农业大学园艺学院,兰州730070

出　　处：《园艺学报》2016年第10期1891-1902,共12页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(31460500)

摘　　要：以‘宝石无核’（Ruby Seedless）葡萄试管苗为材料,采用RT-PCR技术,克隆得到8个葡萄Sn RK2家族基因。序列分析发现,该家族基因蛋白结构在N端相对保守,而在C端极其特异;聚类结果表明葡萄Sn RK2基因家族可以分为3个亚家族;对这8个基因所编码的蛋白质进行分析发现,其富含酸性氨基酸,且均为亲水蛋白;基因组结构分析发现Vv Sn RK2.2和Vv Sn RK2.8含有10个外显子,其他6个均含9个外显子;对蛋白二级结构分析发现8个基因编码的蛋白主要以α–螺旋、β–转角和不规则卷曲为主;亚细胞定位预测,8个基因主要定位于细胞质中。顺式作用元件分析表明,除Vv Sn RK2.1、Vv Sn RK2.2、Vv Sn RK2.6外,其他基因顺式作用元件包含ABRE、DRE/CRT、LRTE中的一个或多个。定量PCR分析表明,Vv Sn RK2的表达存在组织差异性,Vv Sn RK2.7在根中表达水平最高,是叶片的3.8倍,Vv Sn RK2.8在茎中表达水平最高,是叶片的5.0倍。0～–4℃处理后,表达水平下调幅度最小的为Vv Sn RK2.2,Vv Sn RK2.7下调幅度较大,Vv Sn RK2.8的表达水平为0;30℃处理后Vv Sn RK2.2和Vv Sn RK2.5上调表达,分别为对照的3.8倍和3.6倍;Vv Sn RK2.1和Vv Sn RK2.2与盐胁迫调节紧密相关,Vv Sn RK2.5与干旱胁迫调节密切相关。Full length cDNA sequence of 8 grape SnRK2 genes（VvSnRK2）were cloned from grapevine Ruby Seedless using RT-PCR,sequence analysis showed that the N-terminal domain of Vv SnRK2 was highly conserved,while divergent in the C-terminal. The phylogenyetic analysis showed that VvSnRK2 s could be divided into three groups. The Vv SnRK2 s protein carry all acidic amino acids and they all hydrophilic proteins. All VvSnRK2 but VvSnRK2.2,VvSnRK2.8 genes carry 9 exons. The predicted secondary structure suggested that the main structure of eight proteins are alpha helix,beta turn and random coil. The subcellular localization were predicted in the cytoplasmic. The analysis of cis-regulatory elements showed that VvSnRK2 s contained one or more of ABRE,DRE/CRT,LRTE elements except VvSnRK2.1,VvSnRK2.2 and Vv SnRK2.6. Quantitative real-time PCR data indicate that VvSnRK2.7 and Vv SnRK2.8 were expressed at the highest-level in the roots and stems,which were as 3.8 and 5.0 times as leaves,respectively. Under the 0 ℃ and–4 ℃ treatments,VvSnRK2.2 was down-regulated,while the expression was 0 of VvSnRK2.8. Under the 30 ℃ treatment, VvSnRK2.2 and VvSnRK2.5 were up-regulated 3.8 and 3.6 folds,respectively. VvSnRK2.1 and VvSnRK2.2 were response to salt stress,while VvSnRK2.5 was highly response to drought stress.

关 键 词：葡萄 SnRK2家族 基因克隆 荧光定量 基因表达 

分 类 号：S663.1[农业科学—果树学]