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从进境集装箱残余物——小麦麦秆碎片中截获了一种腥黑粉病菌冬孢子,其形态特征和小麦印度腥黑粉病菌冬孢子相似。利用套式PCR和实时荧光PCR分别对截获的冬孢子进行扩增检测,并对扩增产物进行测序和序列比对分析。结果表明:小麦印度腥黑粉病菌特异性引物I9/I4能够扩增截获的腥黑粉病菌冬孢子得到预期的680bp扩增条带,其特异性探针TIND对截获的腥黑粉病菌冬孢子表现为阳性扩增反应;截获的腥黑粉病菌冬孢子ITS区序列和特异性引物PCR扩增产物序列与小麦印度腥黑粉病菌的相应序列完全一致。因此,将截获的腥黑粉病菌冬孢子鉴定为小麦印度腥黑粉病菌Tilletia indica。
From entering the container residue - wheat straw fragments intercepted a teliospore teliospores, its morphological characteristics and wheat Indian black fungus winter spores similar. The intercepted teliospores were amplified by nested PCR and real-time fluorescence PCR, and the amplified products were sequenced and compared by sequence analysis. The results showed that the specific probe I9 / I4 of T. gonorrhoeae could amplify the telomere of intercepted S. gonorrhoeae and obtain the expected 680 bp amplification band. The specific probe TIND was used to detect teliospore Which showed a positive amplification reaction. The intercepted S. cerevisiae ITS region sequence and the specific primer PCR amplification product sequence were exactly the same as the corresponding sequences of wheat Saccharomyces cerevisiae. Therefore, the captured T. gondii teliospores were identified as Tilletia indica wheat.