PLD途径合成的PA增强高表达转玉米C_4型pepc水稻悬浮细胞中PEPC酶活性  

PA Produced by PLD Enhanced PEPC Enzyme Activity of Transgenic Rice Suspension Cell with Over-expressing Maize C_4 Type pepc

作　　者：王满[1] 钱宝云[1] 李霞[1] 

机构地区：[1]江苏省农业科学院粮食作物研究所,江苏省优质水稻工程技术研究中心,南京210014

出　　处：《基因组学与应用生物学》2013年第1期22-29,共8页Genomics and Applied Biology

基　　金：国家自然科学基金项目(No.30871459);江苏省自主创新课题CX(11)1020资助

摘　　要：为了研究外源信号分子对高表达玉米C_4型pepc酶活性的调节机制,本实验以高表达玉米C_4型pepc水稻(PC)和原种水稻Kitaake(WT)的悬浮细胞系为材料,研究葡萄糖、胡蜂蜂毒肽(mastoparan,Mas)及其磷脂酸(phosphatidicacid,PA)的两个合成途径的专一性抑制剂(磷脂酶D(phospholipaseD,PLD)的专一性抑制剂正丁醇和磷脂酶C(phospholipaseC,PLC)的专一性抑制剂新霉素)对供试材料PEPC活性的影响。结果表明,3%葡萄糖处理0.5h对WT的PEPC酶活性抑制最明显;相对于PC,则5%的葡萄糖处理5h时,对其PEPC酶活性抑制的最明显。5mmol/L的Mas对WT和PC悬浮细胞的PEPC酶活性分别抑制了约60%和40%。0.01mmol/L、0.1mmol/L和1mmol/L新霉素处理对WT的PEPC酶活性影响不明显,且无时间效应;与WT相比较,不同浓度的新霉素均增强了PC的PEPC酶活性,其中0.1mmol/L新霉素处理增加为对照的约7倍。正丁醇处理明显抑制了WT的PEPC酶活性,但不同浓度处理之间却没有显著差异;而对于PC,0.02%、0.04%正丁醇处理使得PEPC酶活性显著下降,且比WT的更显著。同时0.04%的正丁醇的抑制效应具有明显的时间效应。以上结果表明,对PC的PEPC酶活性的调节主要依赖于由PLD途径产生的PA的参与。In order to investigate the regulatory mechanism of external signaling molecules on the over-expressing C4-pepc activity, suspension cell lines of over-expressing maize pepc （PC） and Kitaake （WT） were used as materials, and the effects of glucose, mastoparan （Mas） and the two specificity inhibitors of phosphatidic acid （PA） synthesis methods （1-butanol, the specificity inhibitor ofphospholipase D （PLD）; and neomycin, the specificity inhibitor ofphospholipase C （PLC）） on PEPC activity were investigated at the cellular level. The results indicated that there was an obvious inhibition on PEPC activity of WT with the treatment of 3% glucose for 0.5 h; for PC, there was the most notable inhibition with 5% glucose treatment for 5 h. The PEPC activity of both WT and PC could be sup- pressed by 60% and 40% with the 5 mmol/L Mas treatment, respectively. There was no obvious effects on the PEPC activity of WT suspension cells at the different concentrations of neomycin, neither did the effects by time; Compared to WT, the PEPC activity of PC suspension cells was enhanced by neomycin and it was nearly 7 times of controls with 0.1 mmol/L neomycin treatment. The PEPC activity of WT was significantly suppressed by 1-butanol, while it did not have remarkable differences among different concentrations; for PC suspension cells, the treatments of 0.02% and 0.04% 1-butanol made the PEPC activity decreased, which were more obviously than WT. Meanwhile, the inhibition of PEPC activity under the treatment of 0.04% 1-butanol also increased with the treated time. All above results suggest that the regulation to PEPC activity of PC is mainly dependent on the participation of PA by PLD.

关 键 词：水稻 玉米C4型pepc 磷脂酸 磷脂酶C 磷脂酶D 悬浮细胞 

分 类 号：S511[农业科学—作物学]