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以抗白粉病的小麦-长穗偃麦草异代换系山农551为材料,在接种小麦白粉病菌48h制备样品电子显微镜观察,结果表明山农551在白粉病菌侵染早期抑制孢子萌发和菌丝生长.在接种病菌48h后提取RNA,利用cDNARDA和RACE技术克隆WRP1和RPW2共2个基因.BLAST分析表明该二基因均是尚未被克隆的新基因.WRP1没有大的ORF;RPW2的翻译产物含有保守的氨基转移酶结构域,与多种生物的磷酸丝氨酸氨基转移酶有同源序列,可以认为是一新的磷酸丝氨酸氨基转移酶基因.Northern杂交结果显示在小麦白粉病侵染早期RPW2在山农551的叶片中就开始表达.RPW2探针同山农551及其亲本鲁麦5号、济南13和长穗偃麦草基因组DNA进行Southern杂交结果显示,探针与鲁麦5号和济南13无杂交信号,而与山农551和长穗偃麦草都有较强的杂交信号,这表明RPW2来源于长穗偃麦草基因组.
The results showed that Shannong 551 could inhibit spore germination and hyphae during the early stage of powdery mildew inoculation with wheat powdery mildew-resistant wheat cultivar Shannong 551 as material. RNA was extracted 48h after the inoculation of bacteria, and two genes of WRP1 and RPW2 were cloned by cDNARDA and RACE techniques. BLAST analysis showed that the two genes were all new genes that had not been cloned. WRP1 had no large ORF; the translation product of RPW2 contained The conserved aminotransferase domain, which is homologous to a variety of organism phosphoserine aminotransferases, can be considered as a novel phosphoserine aminotransferase gene.Northern hybridization results show that in the early stages of wheat powdery mildew infection RPW2 is in the mountains The expression of RPW2 probe with Shannong551 and its parents Lumai5, Jinan13 and Agropyron elongatum genomic DNA showed that the probe hybridized with Lumai 5 and Jinan 13 without However, the signal of hybridization with Shannong 551 and Eriosphae alba was stronger, indicating that RPW2 was derived from the genome of Eriosphaeaceae.