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利用已获得的喀西茄(Solanum aculeatissimum)接种黄萎病前后的根部转录组数据库,前期筛选得到了4个与植物WRKY转录因子蛋白一致性较高的Unigene序列。系统进化分析表明,这4个转录因子均含有典型的WRKY结构域,分别属于WRKY蛋白家族的第IC类、IIa类和III类。利用实时荧光定量PCR技术分析了这4个WRKY基因在喀西茄不同组织(根、茎和叶)以及接种黄萎病后根部不同时期的表达情况,结果表明,Unigene6502和Unigene20920在根部的表达量显著高于茎部和叶片,而CL1273.Contig2和CL3641.Contig1在叶片的表达量显著高于根部和茎部。这4个WRKY基因在喀西茄根部接种黄萎病菌后具有不同的表达模式,Unigene6502和Unigene20920接种后呈先上调后下调的表达模式,CL1273.Contig2和CL3641.Contig1分别呈上调和下调的表达模式。
Using the obtained Solanum aculeatissimum to inoculate the root transcriptome database of Verticillium dahliae, four Unigene sequences with high identity with plant WRKY transcription factor were obtained. Phylogenetic analysis showed that all four transcription factors contained a typical WRKY domain, belonging to the ICR, IIa and III groups of the WRKY protein family, respectively. The expression of these four WRKY genes in different tissues (roots, stems and leaves) and roots at different stages of Verticillium wilt were analyzed by real-time fluorescence quantitative PCR. The results showed that the expression of Unigene6502 and Unigene20920 at the root Significantly higher than the stems and leaves, while CL1273.Contig2 and CL3641.Contig1 expression in the leaves was significantly higher than the roots and stems. The four WRKY genes had different expression patterns after Verticillium dahliae inoculation in roots of Camellia sinensis, which were up-regulated and down-regulated after Uni6502 and Unigene20920 inoculation, CL1273.Contig2 and CL3641.Contig1 showed up-regulated and down-regulated expression patterns .