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稻瘟病是一种严重危害水稻生产的真菌病害,早期监测和防治是关键。温室接种大田主栽品种合系39和感病水稻蒙古稻,定时观察发病症状。提取水稻病样总DNA,根据稻瘟菌28S rDNA基因序列,设计特异引物和TaqMan探针,进行荧光定量PCR检测。结果表明稻瘟菌在蒙古稻和大田主栽品种合系39上症状最初出现时间为接种后72 h,蒙古稻典型症状出现时间为接种后168 h,在合系39上出现典型症状的时间为接种后190 h以后。利用TaqMan探针荧光定量PCR技术,在接种12 h的蒙古稻和合系39上均能检测到稻瘟菌DNA,接种48 h,菌量拷贝数达到最高峰,接种72 h,菌量拷贝数开始下降。不同品种中病原菌拷贝数存在差别,在接种12 h的蒙古稻中稻瘟菌含量为7.2×103个拷贝数,在合系39中为4.9×103个拷贝数。本研究结果表明,应用实时荧光定量PCR技术可以在接种后12 h症状未显现之前检测到稻瘟菌,为稻瘟病流行监测和早期防治提供了科学依据。
Rice blast is a fungal disease that seriously damages rice production. Early monitoring and control are the key issues. Inoculation of greenhouse cultivars in the main varieties 39 and susceptible rice Mongolia, regular observation of the onset of symptoms. According to the sequence of 28S rDNA of Magnaporthe grisea, specific primers and TaqMan probes were designed and used for quantitative PCR. The results showed that the initial symptoms of M. grisea in Mongolian rice and the main cultivars were 72 h after inoculation, typical symptoms of Mongolian rice appeared 168 h after inoculation, 190 h after inoculation. The DNA of Magnaporthe grisea was detected by TaqMan probe fluorescence quantitative PCR at 12 h after inoculation. After 48 h of inoculation, the number of copies of Magnaporthe grisea reached its peak, and the number of copies reached at 72 h decline. There were differences in the number of copies of pathogens in different varieties, the amount of Magnaporthe grisea was 7.2 × 103 copies in Mongolian rice at 12 h after inoculation, and 4.9 × 103 copies in combined 39. The results of this study show that the application of real-time fluorescence quantitative PCR can detect M. grisea 12 hours after inoculation, and provide a scientific basis for the epidemic surveillance and early prevention of blast.