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水稻鞘氨醇-1-磷酸裂解酶基因OsSPL1的分子特征及其在抗病反应中作用的功能分析

Molecular characterization of rice sphingosine-1-phosphate lyase gene OsSPL1 and functional analysis of its role in disease resistance response.

作者信息

Zhang Huijuan, Jin Xiaoyi, Huang Lei, Hong Yongbo, Zhang Yafen, Ouyang Zhigang, Li Xiaohui, Song Fengming, Li Dayong

机构信息

State Key Laboratory of Rice Biology, Institute of Biotechnology, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, Zhejiang, 310058, People's Republic of China.

出版信息

Plant Cell Rep. 2014 Oct;33(10):1745-56. doi: 10.1007/s00299-014-1653-0. Epub 2014 Aug 12.

Abstract

Our results indicate that overexpression of OsSPL1 in transgenic tobacco plants attenuated disease resistance and facilitated programmed cell death. Long-chain base phosphates including sphingosine-1-phosphate have been shown to act as signaling mediators in regulating programmed cell death (PCD) and stress responses in mammals. In the present study, we characterized a rice gene OsSPL1, encoding a putative sphingosine-1-phosphate lyase that is involved in metabolism of sphingosine-1-phosphate. Expression of OsSPL1 was down-regulated in rice plants after treatments with salicylic acid, benzothiadiazole and 1-amino cyclopropane-1-carboxylic acid, but was induced by infection with a virulent strain of Magnaporthe oryzae, the causal agent of rice blast disease. Transgenic tobacco lines with overexpression of OsSPL1 were generated and analyzed for the possible role of OsSPL1 in disease resistance response and PCD. The OsSPL1-overexpressing tobacco plants displayed increased susceptibility to infection of Pseudomonas syringae pv. tabaci (Pst), the causal agent of wildfire disease, showing severity of disease symptom and bacterial titers in inoculated leaves, and attenuated pathogen-induced expression of PR genes after infection of Pst as compared to the wild-type and vector-transformed plants. Higher level of cell death, as revealed by dead cell staining, leakage of electrolyte and expression of hypersensitive response indicator genes, was observed in the OsSPL1-overexpressing plants after treatment with fumonisin B1, a fungal toxin that induces PCD in plants. Our results suggest that OsSPL1 has different functions in regulating disease resistance response and PCD in plants.

摘要

我们的结果表明,转基因烟草植株中OsSPL1的过表达减弱了抗病性并促进了程序性细胞死亡。包括鞘氨醇-1-磷酸在内的长链碱基磷酸盐已被证明在调节哺乳动物的程序性细胞死亡(PCD)和应激反应中作为信号介质发挥作用。在本研究中,我们鉴定了一个水稻基因OsSPL1,它编码一种推定的鞘氨醇-1-磷酸裂解酶,参与鞘氨醇-1-磷酸的代谢。用水杨酸、苯并噻二唑和1-氨基环丙烷-1-羧酸处理后,水稻植株中OsSPL1的表达下调,但被稻瘟病病原菌稻瘟病菌的强毒株感染所诱导。构建了OsSPL1过表达的转基因烟草株系,并分析了OsSPL1在抗病反应和PCD中的可能作用。与野生型和载体转化植株相比,过表达OsSPL1的烟草植株对接种野火病病原菌丁香假单胞菌烟草致病变种(Pst)的感染敏感性增加,接种叶片上的病害症状严重程度和细菌滴度更高,且在感染Pst后病原菌诱导的PR基因表达减弱。在用伏马菌素B1(一种诱导植物PCD的真菌毒素)处理后,在过表达OsSPL1的植株中观察到更高水平的细胞死亡,这通过死细胞染色、电解质渗漏和过敏反应指示基因的表达得以揭示。我们的结果表明,OsSPL1在调节植物的抗病反应和PCD中具有不同的功能。

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