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根据稻曲病菌核糖体转录间隔区(ITS)序列的特异性,设计了2对引物及相应TaqMan探针用于建立稻曲病菌实时PCR定量检测技术。结果表明,uvr292rf/uvr397rr/uvrp333组合建立的体系可特异性检测出稻曲病菌,扩增基线平整,指数扩增期明显,重现性好,最低可检出24fg的稻曲病菌基因组DNA模板量;以梯度稀释分生孢子液提取的基因组DNA为模板,建立孢子数量常用对数值与Ct值的线性关系,理论上最低可检测出0.67个分生孢子。对3个时间点的田间孢子捕捉样品进行检测,结果显示不同月份间稻曲病菌孢子数量存在差异。
Two pairs of primers and corresponding TaqMan probes were designed to establish the real-time PCR quantitative detection technology of V. tumefaciens based on the ITS sequence of the rice false smut. The results showed that the system established by the combination of uvr292rf / uvr397rr / uvrp333 could specifically detect the strains of U. virens, the baseline of amplification was flat, the exponential phase of amplification was obvious and the reproducibility was good. The lowest detectable 24fg of genomic DNA template The linear relationship between the logarithmic value of the number of spores and the Ct value was established by using the genomic DNA extracted by the gradient dilution of the conidia liquid as a template. In theory, a minimum of 0.67 conidia could be detected. The spore-captured samples at three time points were tested, and the results showed that there were differences in the number of spores between the two strains in different months.