《植物生理学报》 2018, 54(1): 157-164

霸王环核苷酸门控通道基因ZxCNGC5的克隆及表达模式分析

刘亚琪, 王文颖, 崔彦农, 郭欢, 李孟湛, 王锁民^*

兰州大学草地农业科技学院, 草地农业生态系统国家重点实验室, 兰州730020

通信作者：王锁民；E-mail: smwang@lzu.edu.cn

摘　要：

钙离子(Ca²⁺)作为生命活动过程中重要的第二信使, 对植物的生长发育至关重要。环核苷酸门控离子通道(cyclic nucleotide-gated channels, CNGCs)是一类Ca²⁺通道, 在植物的花粉发育、免疫反应以及响应逆境胁迫等生理过程中发挥着重要作用。本研究通过RACE和RT-PCR技术克隆了荒漠旱生植物霸王(Zygophyllum xanthoxylum)CNGCs家族的一个编码基因, 命名为ZxCNGC5。生物信息学分析显示其编码728个氨基酸, 包含CNGCs家族所特有的6个跨膜域、1个P环以及环核苷酸结合域(cyclic nucleotide-binding domain, CNBD)和钙调蛋白结合域(calmodulin-binding domain, CaMBD); 系统进化树将CNGCs家族分为5个亚族(I、II、III、IVA和IVB), ZxCNGC5属于第II亚族; 组织特异性分析显示该基因主要在根中表达; 在0.01和10 mmol·L^-1 CaCl₂处理下, ZxCNGC5在根中的表达量短期内均显著升高, 并分别在3和12 h达到最高, 随后逐渐降低, 表明ZxCNGC5在根中的表达受外界CaCl₂的短期诱导。

关键词：霸王; Ca²⁺; CNGC5; 基因克隆; 表达模式

收稿：2017-10-11   修定：2017-12-22

资助：国家自然科学基金(31730093和31470503)。

Cloning and expression analysis of cyclic nucleotide-gated channels gene ZxCNGC5 from Zygophyllum xanthoxylum

LIU Ya-Qi, WANG Wen-Ying, CUI Yan-Nong, GUO Huan, LI Meng-Zhan, WANG Suo-Min^*

State Key Laboratory of Grassland Agroecosystems, College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou 730020, China

Corresponding author: WANG Suo-Min; E-mail: smwang@lzu.edu.cn

Abstract:

Ca²⁺, a critical second messenger in the processes of plant life, is essential for the growth and development of plants. Cyclic nucleotide-gated channels (CNGCs) are Ca²⁺-permeable channels that play an important role in pollen tube guidance, responses to biotic and abiotic stresses as well as immune responses. Here, by RACE and RT-PCR technology, we cloned a new CNGC from Zygophyllum xanthoxylum, designated as ZxCNGC5. By bioinformatics analysis of the ZxCNGC5, we found that this gene encoded 728 amino acids, including six transmembrane helix with a Pore loop, cyclic nucleotide-binding domain and calmodulin-binding domain, which were unique to CNGCs family. Phylogenetic analysis showed that CNGCs family could be divided into 5 subfamily (I, II, III, IVA and IVB) and ZxCNCG5 belonged to the II subfamily. Tissue specificity analysis suggested that the gene was especially expressed in roots. Further, under the 0.01 and 10 mmol·L^-1 CaCl₂ treatment, the expression of ZxCNGC5 in roots was significantly induced in short term and reached the highest at 3 and 12 h, and then decreased gradually. These results indicated that the expression of ZxCNGC5 in roots was induced in short time when adding the CaCl₂.

Key words: Zygophyllum xanthoxylum; Ca²⁺; CNGC5; gene cloning; expression pattern