Guo Guan-Ying, Li Ming-Jie, Wang Peng-Fei, Wang Feng-Qing, He Hua-Qin, Li Juan, Zheng Hong-Yan, Chen Xin-Jian, Zhang Zhong-Yi
School of Agronomy, Henan Agricultural University, Zhengzhou 450002, China.
Zhongguo Zhong Yao Za Zhi. 2013 May;38(10):1471-8.
Based on the early transcriptome and digital differentially expressed profiling library construction in consecutive monoculture (two-year culturing) Rehmannia glutinosa, we screened and chose the twelve differentially expressed protein genes which might be related with calcium signal system. The spatiotemporal expression of these genes was measured by the real-time quantitative PCR, and the relative expression values of the genes related with calcium signal system in different development stages and tissues of normal growth (one-year culturing) and succession cropping of R. glutinosa (two-year culturing) was elaborated in detail. In addition, disposed succession cropping of R. glutinosa was treated with different levels of calcium signal blocking agents in order to verify the mode of action of calcium signal system on consecutive monoculture problem in R. glutinosa. Among the twelve genes, two calcium channels away from the cytoplasm were down-regulated expressed, while the ten calcium channels toward the cytoplasm were up-regulated expressed. The results implied that succession cropping caused calcium ions flowing from endoplasmic reticulum to cytoplasm. While the key genes in calcium signal respond components such as CBL, CBP, CIBP, PLC, etc. were down-regulated expressed significantly in succession cropping of R. glutinosa which were disposed with calcium signal blocking agents, the extent of the damage was relieved, and approached the normal growth (one-year culturing) level. This result strongly showed that calcium signal system participated in the perceiving, conducting and magnifying processes of succession cropping obstacles of R. glutinosa.
基于地黄连续单作（两年栽培）早期转录组及数字差异表达谱文库构建，我们筛选并选取了12个可能与钙信号系统相关的差异表达蛋白基因。通过实时定量PCR测定这些基因的时空表达，并详细阐述了正常生长（一年栽培）和地黄连作（两年栽培）不同发育阶段及组织中与钙信号系统相关基因的相对表达值。此外，对地黄连作进行处理，用不同水平的钙信号阻断剂处理，以验证钙信号系统对地黄连作问题的作用模式。在这12个基因中，两个远离细胞质的钙通道下调表达，而10个朝向细胞质的钙通道上调表达。结果表明，连作导致钙离子从内质网流向细胞质。而在经钙信号阻断剂处理的地黄连作中，钙信号响应元件如CBL、CBP、CIBP、PLC等的关键基因显著下调表达，损伤程度减轻，并接近正常生长（一年栽培）水平。这一结果有力地表明，钙信号系统参与了地黄连作障碍的感知、传导和放大过程。
