纹枯病菌诱导水稻表达的WRKY基因克隆及分析

摘要

[目的]克隆纹枯病菌诱导的水稻上调表达基因.[方法]对应用抑制差减杂交枝术(SSH)获得的纹枯病菌诱导水稻表达的EST序列K16进行电子克隆,根据电子克隆产物设计引物进行RT-PCR、克隆测序,利用生物信息学技术对克隆产物进行功能预测.[结果]克隆出一个长度为1079bp的序列,结构功能分析显示该基因编码236个氨基酸的蛋白,存在多个功能位点,含有2个典型的WRKY结构域和1个C2H2锌指型结构,该基因与水稻WRKY8、WRKY24和WRKY30基因有较高的同源性.[结论]经电子克隆获得的纹枯病菌诱导表达的基因为典型的WRKY家族基因,该基因可能在水稻抗纹枯病中起重要作用.%[ Objective ] The aim was to clone the up-regulated expression gene of rice induced by Rhizoctonia solani. [ Method ] The EST fragment K16 obtained by suppression subtraction hybridization (SSH) was cloned and confirmed by reverse transcription-polymerase chain reaction (RT-PCR). Then RT-PCR products were cloned into the PMD18-T vector and sequenced. The functions of the sequence were predicted with bioinformatics method. [ Result] A 1 079 bp gene was obtained. The gene encoded a protein with 236 amino acids. The protein contains many motif sites, two WRKY domains and a C2H2 zinc finger motif. The gene showed high identities with WRKY8, WRKY24 and WRKY30 gene of rice. [ Conclusion] The up-regulated expression gene induced by R. solani was representative WRKY family gene. The gene could play an important role on rice sheath blight resistance.