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为了阐明Rboh基因家族在水稻免疫应答中的功能,首先利用生物信息学方法从水稻基因组数据库中检索到7个水稻Rboh基因,并对其进行系统发育学和组织特异性表达分析,发现OsRbohD只在穗和愈伤组织中表达,且OsRbohE和OsRbohF仅在愈伤组织中特异表达,而其他基因为组成型表达。利用Real-time PCR对分别在水杨酸SA、茉莉酸甲酯MeJA和水稻黄单胞菌PXO99致病菌株处理下的OsRboh基因家族的表达水平进行了分析,同时对处理后的叶片H2O2含量进行测定。结果表明,SA可以提高OsRbohA、OsRbohB、OsRbohC和OsRbohD的表达水平,MeJA可以提高OsRbohA、OsRbohB、OsRbohC和OsRbohG的表达水平,接种水稻黄单胞菌致病菌株PXO99可以提高OsRbohA和OsRbohB的表达水平。然而三者在诱导OsRboh基因家族成员的表达时序性和表达程度存在着差异。此外,3种不同处理都能导致不同程度的H2O2积累。结果显示,OsRboh基因家族各基因在水稻免疫应答中可能扮演不同角色,发挥不同作用。
In order to elucidate the function of Rboh gene family in rice immune response, seven rice Rboh genes were retrieved from rice genome using bioinformatics methods. Phylogenetic and tissue-specific expression analysis of Rboh gene revealed that OsbobohD Spike and callus, and OsRbohE and OsRbohF were exclusively expressed in callus while other genes were constitutively expressed. The expression levels of OsRboh gene family under salicylic acid SA, methyl jasmonate MeJA and PXO99 pathogenic strains were analyzed by Real-time PCR, and the content of H2O2 in the treated leaves was also analyzed Determination. The results showed that SA could increase the expression levels of OsRbohA, OsRbohB, OsRbohC and OsRbohD. MeJA increased the expression levels of OsRbohA, OsRbohB, OsRbohC and OsRbohG. The inoculation of PXO99 caused the increase of OsRbohA and OsRbohB expression. However, there are differences in the timing and expression of OsRboh gene family between the three groups. In addition, all three treatments resulted in different levels of H2O2 accumulation. The results showed that OsRboh gene family genes may play different roles in rice immune response and play different roles.