建立一种烟草根黑腐病菌新靶标的分子检测方法。以β-微管蛋白基因为靶标设计了特异性引物Tbas-tub2F/Tbas-tubR用于根黑腐病菌的PCR检测，对其检测特异性、灵敏度、早期诊断技术进行了测试。结果显示，只有根黑腐病菌能扩增到一条254 bp的特异性条带，其他19种真菌、卵菌及阴性对照均无扩增产物，表明该对引物能特异性地检测根黑腐病菌。该引物对根黑腐病菌基因组DNA检测的灵敏度为10 fg/μL。Tbas-tub2F/Tbas-tubR可从接种侵染24 h后的烟草组织中检测到根黑腐病菌，从而对根黑腐病进行准确的早期诊断。开发了烟草根黑腐病菌新靶标的分子检测和早期诊断技术。%To establish a molecular detection method of new target for tobacco black root rot caused by Thielaviopsis basicola, β-tubulin gene was selected as a new target and a pair of species-specific primers Tbas-tub2F /Tbas-tubR was designed to detect T. basicola by PCR. The specificity, sensitivity and early diagnosis technology of the primers were assayed. The results showed that an expected 254 bp band could be amplified only from T. basicola and no PCR product was amplified from any of the other nineteen fungi and oomycetes and negative control, which suggested that the primers could be used to detect T. basicola specifically. The detection sensitivity of Tbas-tub2F/Tbas-tubR primers was 10 fg genomic DNA of T. basicola. The pathogen could be detected from tobacco tissues 24 hours post inoculation thus the primers were good for early accurate diagnosis of tobacco black root rot disease. In this study we developed an alternative new target for both molecular detection and early diagnosis of tobacco black root rot caused by T. basicola.
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