研究夏孢子萌发过程的分子机制对于从分子水平上理解条锈菌的侵染过程及其与寄主互作的关系具有重要的理论意义。本研究以小麦条锈菌Puccinia striiformis f.sp tritici32号生理小种(CY32)为材料,研究了用水培方法萌发夏孢子的适宜条件。结果显示,CY32夏孢子萌发的最适温度为9℃,最适宜的孢子量是6mg/200mL水,适宜的溶液是无菌蒸馏水。水化能促进夏孢子的萌发,新鲜夏孢子和干燥容器中放置2d的夏孢子经水化15h后,萌发率显著提高。此方法获得的萌发夏孢子提取的RNA可以满足eDNA文库构建和基因表达分析等分子生物学研究的要求,并为小麦条锈菌的分子生物学研究奠定了物质基础。
It is valuable for molecular biology study of Puccinia striiformis f. sp tritici to obtain masses of germinated urediospores. In this study urediospores of CY32 race were used to explore optimal condition of germination with incubation in water in a large scale. The results demonstrated that the optimal temperature is 9℃. The optimal inoculation dose is 6mg/200mL water. Sterilized distilled water is more beneficial than tap water to germinate. Hydration of urediospores is propitious to germinate. Percent germination of both of fresh and 2d-stored urediospores increased significantly with 15h hydration. Total RNA isolated from geminated urediospores which incubated by this method meet the needs of subsequent molecular biology study such as construction of cDNA library and analysis of gene expression. This study provides solid foundation for future molecular biology research in the obligate plant pathogen Puccinia striiformis f.sp tritici.