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利用小麦离体叶片瞬时表达技术分析了3个小麦抗白粉病相关基因CAF1、BI-1和DAD1的功能。根据小麦响应白粉菌侵染的差异表达基因芯片检测结果,筛选出表达差异的抗病相关基因CAF1和BI-1,并挑选与植物程序性死亡相关的基因DAD1,将克隆到的这3个小麦基因全长ORF序列构建入超表达载体pAHC25中,利用瞬时表达技术检测目标基因和GUS基因共转化的阳性小麦叶片表皮细胞中白粉菌成功侵入并形成吸器的细胞比例(侵入频率),研究目标基因的超表达后对白粉菌入侵及吸器形成产生的影响。结果表明,小麦CAF1基因在感病小麦品种叶片中的瞬时表达,对白粉病侵入和吸器形成具有一定的抑制作用(侵入频率为(37.37±3.2)%,对照为(54.75±0.6)%),在一定程度上增强了表达细胞对白粉菌的抗性;表达DAD1基因则可能在一定程度上有利于白粉菌吸器的形成(侵入频率为(57.34±1.1)%,对照为(50.93±1.3)%);而在感病叶片中表达基因BI-1对白粉菌吸器形成没有明显影响(侵入频率为(54.31±1.7)%,对照为(52.37±1.8)%)。
The function of three wheat resistant genes for powdery mildew-associated genes CAF1, BI-1 and DAD1 was analyzed using the transient expression of wheat leaves in vitro. According to the differential expression gene chip detection results of wheat infected with powdery mildew, the expression-related disease-resistant genes CAF1 and BI-1 were screened out and the gene DAD1 related to plant apoptosis was selected. The cloned wheat The full-length ORF sequence of the gene was constructed into the overexpression vector pAHC25. The transient expression of target gene and GUS gene co-transformed positive wheat leaf epidermal cells were used to detect the percentage of invasive cells that formed the aspirate. Of the over-expression of powdery mildew invade and the formation of the impact of inhaler. The results showed that the transient expression of wheat CAF1 gene in the leaves of susceptible wheat cultivars had an inhibitory effect on invasion and inhalation of powdery mildew (37.37 ± 3.2% in control and (54.75 ± 0.6)% in control) The expression of DAD1 gene may be beneficial to the formation of powdery mildew inhaler to a certain extent (the frequency of invasion is (57.34 ± 1.1)%, the control is (50.93 ± 1.3)%) ). However, the expression of BI-1 in susceptible leaves had no significant effect on the formation of powdery mildew (54.31 ± 1.7%) and (52.37 ± 1.8)%, respectively.