葡萄ZTL、COP1基因的鉴定及其在不同光质和转光条件下的表达分析  

Identification of the ZTL and COP1 gene in Vitis vinifera and expression analysis of different light qualities and light transfers

作　　者：马彦妮 陈佰鸿[1] 毛娟[1] 左存武[1] 吴金红[1] 杨始锦 胡紫璟 李婷婷[1] 

机构地区：[1]甘肃农业大学园艺学院,兰州730070

出　　处：《果树学报》2017年第5期541-551,共11页Journal of Fruit Science

基　　金：国家自然科学基金(31401500)

摘　　要：【目的】在葡萄试管苗离体培养下,研究不同波长对基因ZTL、COP1表达的影响。【方法】以‘红地球’葡萄(Vitis vinifera‘Red Globe’)试管苗为材料,经RT-PCR法克隆获得葡萄ZTL、COP1基因全长cDNA,并利用生物信息学分析了以上基因的理化性质、蛋白质二级结构、亚细胞定位以及系统进化树。利用q RT-PCR技术分析了以上基因在不同光质处理下表达量的变化情况。利用荧光参数系统分析不同光质对葡萄试管苗生长的影响。【结果】ZTL的分子式为C1972H3107N553O576S20,是亲水性稳定蛋白,脂溶性较差;COP1的分子式为C832H1291N209O225S9,是疏水性稳定蛋白,脂溶性较好。ZTL和COP1的二级结构均由α-螺旋、β-转角、无规则卷曲和延伸链结构组成;亚细胞定位分析结果显示,ZTL、COP1均位于细胞质中;系统进化分析表明,ZTL与桃的亲缘关系最近,COP1与其他物种的亲缘关系均较远,单独聚为一个亚族。在不同光质处理后ZTL均为上调表达,其中白光转蓝光、红光、红光转白光、红光转蓝光和蓝光转红光处理后分别上调达11.2、22.3、12.0、32.9和19.6倍。不同光质处理后COP1基因的表达呈现不同的变化趋势:红光转蓝光处理后,COP1上调1.97倍,红光处理后表达量无显著变化,其他光质处理后均为下调表达,其中蓝光转白光和蓝光转红光处理后下调达18倍。荧光参数分析表明,qP、qN、NPQ和F_v/F_m在转光培养中表达活跃,均在红光转蓝光的转光培养下表达量最高。【结论】ZTL在葡萄转光培养中表达活跃,均呈显著上调趋势,而COP1总体表现为下调趋势。[ Objective ]The ZTL and COP1 genes in Vitis vinifera were identified. Furthermore, the expres- sion levels were detected after the plantlets were treated with different light conditions. [Methods ] Full- length cDNA sequences of ZTL and COP1 were successfully cloned from tissue cultural plantlets of the ＇Red Globe＇ grape （Vitis vinifera）. To understand their structure and function, physical and chemical characterization, protein secondary structure, sub-cellular localization, secondary structure and phyloge- netic relationships of the two genes above were determined by using bioinformatic analysis. The expres- sions of the two genes were analyzed by using qRT-PCR and applying different treatments. In addition, the fluorescence expressions of the grape test-tube plantlets were analyzed by using a fluorescence param- eter and applying different treatments.

关 键 词：葡萄试管苗 ZTL基因 COPI基因 克隆 表达分析 

分 类 号：S663.1[农业科学—果树学]