棉花CML基因家族成员鉴定与功能分析  

Identification and Functional Analysis of CML Gene Family in Cotton

作　　者：杨秀 许艳超 杨芳芳 蔡小彦[1] 侯宇清 王玉红[1] 王星星[1] 王坤波[1] 刘方[1] 周忠丽[1] Xiu;Xu Yanchao;Yang Fangfang;Cai Xiaoyan;Hou Yuqing;Wang Yuhong;Wang Xingxing;Wang Kunbo;Liu Fang;Zhou Zhongli(State Key Laboratory of Cotton Biology/Institute of Cotton Research of Chinese Academy of Agricultural Sciences,Anyang,Henan 455000,China)

机构地区：[1]棉花生物学国家重点实验室/中国农业科学院棉花研究所

出　　处：《棉花学报》2019年第4期307-318,共12页Cotton Science

基　　金：国家重点研发计划(2016YFD0100203,2017YFD0101601)

摘　　要：【目的】类钙调素(Calmodulin-like,CML)蛋白是1种重要的Ca2+传感器,在调节植物应激反应机制中起重要作用。对CML基因家族进行全基因组学分析,以便深入研究CML基因在棉花逆境胁迫下的作用。【方法】利用生物信息学的方法进行了棉花CML家族成员的鉴定。利用转录组数据和反转录聚合酶链式反应分析陆地棉(Gossypium hirsutum L.)CML基因在盐胁迫下的表达模式。利用病毒诱导基因沉默技术对GhCML44-2基因进行沉默并进行功能验证。【结果】在陆地棉(Gossypium hirsutum L.)、雷蒙德氏棉(G. raimondii Ulbrich)、亚洲棉(G. arboreum L.)中分别获得了154个、74个、78个CML蛋白。进化树和保守基序分析表明,GhCML蛋白分为8个亚类,都含有保守的EF-hand结构域;在盐胁迫下,107个GhCML基因在叶和根中有显著的差异表达,且在根和叶中的差异表达模式不同。启动子分析表明,这些基因启动子中存在多种不同的胁迫响应元件;利用病毒诱导基因沉默技术成功沉默GhCML44-2的棉株相较于对照更加不耐盐。【结论】该研究结果有助于了解棉花CML基因家族的进化与功能,为后续研究其功能提供了一定的理论依据。[Objective] Calmodulin-like protein (CML) is an important Ca2+ sensor and plays an important role in regulating plant stress response mechanism. Genome-wide analysis of CML gene family provides a basis for further study of the role of CML genes in cotton under stress.[Method] The members of CML family in cotton were identified by bioinformatics. The expression patterns of CML genes in upland cotton under salt stress were analyzed by the transcriptome data and real-time reverse transcription-polymerase chain reaction. Virus-induced gene silencing (VIGS) was used to silence GhCML44-2 gene and to verify its function.[Result] 154, 74 and 78 CML proteins were obtained from Gossypium hirsutum L., G. raimondii Ulbrich and G. arboreum L., respectively. Evolutionary tree and conservative motif analysis showed that GhCML proteins were divided into eight subgroups, all containing conserved EF-hand domain. Under salt stress, 107 GhCML genes were differentially expressed with different expression patterns in leaves and roots. Promoter analysis showed that there were many different stress response elements in these gene promoters, and cotton plants were more salt-tolerant than control plants after silencing the expression of GhCML44-2 successfully by using VIGS.[Conclusion] The results of this study are helpful to understand the evolution and function of CML gene family in cotton, and provide a theoretical basis and reference for the follow-up study of their specific functions in cotton.

关 键 词：棉花 类钙调素蛋白 盐胁迫 病毒诱导基因沉默 基因功能 

分 类 号：S562[农业科学—作物学]