青花菜Whirly基因的克隆、系统进化及表达特征分析  

Cloning,phyletic evolution and expression characteristics analysis of Whirly gene in broccoli

作　　者：荆赞革 裴徐梨[1] 李成慧 唐征 冯鹏宇[1] JING Zan-ge;PEI Xu-li;LI Cheng-hui;TANG Zheng;FENG Peng-yu(College of Agriculture and Bioscience,Kunming University,Kunming 650214,China;College of Agriculture and Biotechnology,Wenzhou Vocational College of Science and Technology,Wenzhou,Zhejiang 325006,China)

机构地区：[1]昆明学院农学与生命科学学院,昆明650214 [2]温州科技职业学院农业与生物技术学院,浙江温州325006

出　　处：《西南农业学报》2022年第12期2719-2723,共5页Southwest China Journal of Agricultural Sciences

基　　金：浙江省自然基金项目(LY18C150006);云南省应用基础研究计划项目(2018FH001-044);云南省科技厅基础研究专项青年项目(2019-1-C-25318000002236);浙江省农业(蔬菜新品种选育)新品种选育重大科技专项子项目(2021C02065-4-2);温州市农业新品种选育协作组子项目(2019ZX007-2)。

摘　　要：【目的】克隆青花菜Whirly基因,并分析其序列特征和表达模式,为进一步深入研究其渍水胁迫响应机制及其生物学功能奠定基础。【方法】以“KU20-7”为材料,利用RT-PCR技术克隆得到青花菜Whirly基因,并对其分子量、等电点和二级结构进行分析。利用软件MEGA 6.0对部分植物代表种的Whirly蛋白进行序列比对,而后构建系统进化树。【结果】从青花菜中克隆到Whirly基因家族的3个成员,命名为BoWHY1、BoWHY2和BoWHY3,其基因长度分别为795、723和783 bp,推导分别编码264、240和260个氨基酸。系统进化分析表明,低等植物(藻类、苔藓和裸子植物)Whirly成员位于进化树的底端,被子植物Whirly成员则聚为1个大组。被子植物中乌油樟、葡萄、杨树和木瓜相对独立,挑选的十字花科物种则聚在一起。青花菜BoWHY2与BoWHY1/BoWHY3的亲缘关系相对较远。qRT-PCR分析显示,在渍水胁迫下BoWHY1的表达量随胁迫时间的延长逐渐递减。BoWHY2和BoWHY3的表达量呈现出先上升(2 d)后下降(6 d)的趋势,且BoWHY2的响应更加剧烈。【结论】植物Whirly基因家族可能由一个祖先基因进化而来。在长期进化过程中青花菜旁系同源基因BoWHY1和BoWHY3可能发生了亚功能化,渍水胁迫的表达特征也印证了此观点。【Objective】Whirly gene from broccoli was cloned and sequence characteristic and expression pattern were analyzed in the study,to lay a foundation for further study on its response mechanism of waterlogging stress and biological function.【Method】Using‘KU20-7’as material,Whirly genes were cloned from broccoli by RT-PCR technology,and their molecular weight,isoelectric point and secondary structure of amino acid were also analyzed.The amino acid sequence of Whirly protein from some representative plant species were compared,and then the phylogenetic tree was constructed by MEGA 6.0.【Result】Three members of Whirly gene family were cloned from broccoli,naming BoWHY1,BoWHY2 and BoWHY3.The gene lengths were 795,723 and 783 bp,respectively,and encoding 264,240 and 260 amino acids.Phylogenetic analysis showed that the Whirly members of lower plants(algae,mosses and gymnosperms)were located at the bottom of evolutionary tree,while the Whirly members of angiosperms were grouped into a large group.Among angiosperms,Amborella trichopoda,Vitis vinifera,Populus trichocarpa and Carica papaya were relatively independent,while the selected Cruciferae species were gathered together.The genetic relationship between BoWHY2 and BoWHY1/BoWHY3 was relatively distant in broccoli.qRT-PCR analysis results showed that the expression of BoWHY1 decreased gradually with prolongation of waterlogging stress.While BoWHY2 and BoWHY3 showed a trend of first rising(2 days)and then falling(6 days),and the stress response of BoWHY2 was more intense.【Conclusion】The Whirly gene family may have evolved from an ancestor gene in plants.In the process of long-term evolution,the paralogous genes BoWHY1 and BoWHY3 may have been sub functionalized in broccoli,and their expression characteristics under waterlogging stress also confirmed this view.

关 键 词：青花菜 Whirly 系统进化 渍水胁迫 表达特征 

分 类 号：S635[农业科学—蔬菜学]