地黄钙信号体系相关基因的鉴定及其在连作条件下响应特征分析  

Identification and analysis of the calcium signaling system genes responding to continuous cropping injury in Rehmannia glutinosa L.

作　　者：杨楚韵 李鑫宇 李明杰[1,3] 冯法节 古力[1] 张宝[1] 陈淑强 陈爱国 张重义[1,3] 

机构地区：[1]福建农林大学作物科学学院,福建福州350002 [2]河南农业大学农学院,河南郑州450002 [3]福建农林大学福建农林大学作物遗传育种与综合利用教育部重点实验室,福建福州350002

出　　处：《药学学报》2017年第12期1951-1961,共11页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81573538;81503193;81403042;81603243);福建省教育厅省属高校专项(JK2015013);高等学校博士学科点专项科研基金资助课题(博导类)(20123515110005)

摘　　要：钙信号是植物响应胁迫的重要系统之一,前期研究发现钙信号参与了连作地黄感知、传导和响应连作胁迫的进程。为了确证钙信号在地黄连作障碍形成过程中的作用方式,本研究在前期地黄大容量转录组基础上,利用生物信息学方法详细鉴定地黄钙信号转导体系相关蛋白的编码基因,并对其在连作和头茬地黄中的表达模式进行分析。同时,采用焦锑酸盐法和钙荧光染色法测定连作和头茬地黄根细胞中钙离子浓度差异。鉴定出5个CaMs、12个CBLs、21个CDPKs、21个CIPKs、16个CMLs和9个CRKs等84个钙信号体系相关基因。通过对连作和头茬地黄钙信号体系相关基因差异表达模式进行分析,发现连作效应增强了地黄钙信号体系中关键基因的表达。进一步利用qRT-PCR方法对12个关键的钙信号转导相关基因进行表达验证,确证了钙信号响应了连作胁迫进程。此外,2种钙离子浓度检测方法也均证实连作胁迫显著促进了地黄胞内钙离子的积累。本研究从分子和细胞水平上确证了钙信号在地黄连作障碍形成中扮演着重要角色,为进一步从分子水平上解读地黄连作障碍的形成机制奠定了基础。Calcium signaling plays a critical role in response to various abiotic and biotic stresses in plants.Preliminarily evidence showed that calcium signaling perceived and transduced the harmful signaling generated from continuous cropping stress in R.glutinosa.To investigate the roles of calcium signaling in continuous cropping injury formation,the key genes involved in calcium signaling transduction were identified in R.glutinosa transcriptome through bioinformatic methods.Furthermore,the calcium ion concentration in both normal and continuous cropping R.glutinosa root cells were measured by potassium pyroantimonate precipitation and calcium fluorescence method.As a result,a set of 84 calcium signaling-related genes,including 5 Ca Ms,12 CBLs,21 CDPKs,21 CIPKs,16 CMLs,and 9 CRKs were captured in R.glutinosa transcriptome.The analysis of expression profile in continuous cropping compared to normal growth R.glutinosa indicated that continuous cropping stress significantly increased the expression of calcium signaling-related genes in continuous cropping R.glutinosa.At the same time,the abundance levels of 12 calcium signaling-related genes quantified by q PCR further validated the high expression of calcium signaling-related genes presented in continous cropping R.glutinosa.In addition,the continuous cropping condition significantly promoted the accumulation of intracellular calcium ions in R.glutinosa based on two methods of potassium pyroantimonate precipitation and calcium fluorescence.This study verified the possible roles of calcium signaling in the formation of continuous cropping injury on molecular and cellular level,which lays a solid foundation for illuminating formation mechanism of continuous cropping injury on molecular level.

关 键 词：地黄 连作障碍 自毒作用 钙信号体系 钙离子浓度 

分 类 号：R931[医药卫生—生药学]