山田胶锈菌和亚洲胶锈菌吸器提取体系建立

Construction of haustorial isolation systems of Gymnosporangium yamadae and G. asiaticum

利用锈菌吸器特异性结合伴刀豆球蛋白Concanavalin A(ConA)的特性,用ConA-sepharose 4b作为层析介质,对自然状态下感染山田胶锈菌Gymnosporangium yamadae的绚丽海棠叶片以及接种亚洲胶锈菌G. asiaticum后不同发病时期沙梨叶片中胶锈菌的吸器进行分离,构建了吸器提取体系。结果表明,在感病叶片产生大量花斑时提取到这两种胶锈菌的完整吸器,经光学显微镜与透射电子显微镜观察,发现这两种胶锈菌的吸器形态相似,呈菌丝状或肾形或不规则形状,吸器体与叶绿体含量接近1:1。研究中提取得到了G. yamadae吸器总RNA,为胶锈菌的吸器转录组中候选效应分子筛选以及病原与寄主互作分子机制研究奠定了重要基础。

Based on the property of Concanavalin A(ConA)that specially binds haustoria of rust fungi,ConA-sepharose 4 b was used as affinity chromatography medium to isolate haustoria from Gymnosporangium yamadae infected leaves of Malus‘Radient’under natural conditions and G.asiaticum infected leaves of Pyrus pyrifolia by artificial inoculation at different time.Intact haustoria of G.yamadae and G.asiaticum were successfully isolated from the infected leaves showing chlorotic flecks,respectively.Vast amount of haustoria showing similar shape were observed under light and transmission electron microscopy.The haustoria are mainly haphal,kidney-shaped or irregular.The ratio of haustoria to chloroplast in both rusts was approximately 1:1.Total RNA of the isolated haustoria of G.yamadae were extracted by traditional TRIzol method.This study provided a basis for further investigating on screening candidate effectors from the haustorial transcriptome and molecular mechanism of pathogen-host interaction.