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

发布时间：2018-02-07 14:00

本文关键词： 地黄连作障碍自毒作用钙信号体系钙离子浓度　出处：《药学学报》2017年12期 　论文类型：期刊论文

【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。
[Abstract]:Calcium signal is one of the important systems of plant response to stress. Previous studies have found that calcium signal is involved in the process of perception, conduction and response to continuous cropping stress of Rehmannia glutinosa. In this study, on the basis of large volume transcriptome of Rehmannia glutinosa, we used bioinformatics to identify the encoding genes of the proteins associated with the calcium signal transduction system of Rehmannia glutinosa, and analyzed their expression patterns in continuous cropping and first stubble Rehmannia glutinosa. Calcium concentrations in root cells of continuous cropping and stubble Rehmannia rehmanniae were determined by pyroantimonate method and calcium fluorescence staining. 84 calcium signaling system genes including 5 CaMs, 12 CBLs, 21 CDPKs, 21 CIPKs, 16 CMLs and 9 CRKs were identified. The differentially expressed patterns of genes related to calcium signaling system in Rehmannia glutinosa and Rehmannia glutinosa were analyzed. It was found that continuous cropping enhanced the expression of key genes in the calcium signaling system of Rehmannia glutinosa. The expression of 12 key calcium signal transduction related genes was verified by qRT-PCR method. It was confirmed that the calcium signal responded to the process of continuous cropping stress. In addition, two methods for the determination of calcium concentration also confirmed that continuous cropping stress significantly promoted intracellular calcium accumulation in Rehmannia glutinosa. In this study, the molecular and cellular levels of calcium were confirmed. Is playing an important role in the formation of obstacles to the formation of Rhizoma Coptidis. It lays a foundation for further understanding the formation mechanism of continuous cropping barrier of Rehmannia glutinosa at molecular level.
【作者单位】：福建农林大学作物科学学院;河南农业大学农学院;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;
【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)
【分类号】：S567.234

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