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| Method for Rapid Detection of Fusarium oxysporum andPhytophthora parasitica var. nicotianae in Tabacco Planting Soil |
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| DOI：10.16768/j.issn.1004-874X.2022.02.012 |
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| Abstract: |
| 【Objective】Fusarium wilt and tobacco black shank, caused by Fusarium oxysporum and Phytophthora
parasitica var. Nicotianae, are common soil-borne diseases in tobacco planting soil, leading to severe damages. Therefore,
a rapid detection method for the two pathogens was established to provide technical support for disease epidemic and
treatment in the process of tobacco planting and ensure the stable production of tobacco industry.【Method】Taking the
soil with soil-borne disease and healthy soil in Baise area of Guangxi were used as test materials, and the standard curve
was established with the standard pathogen as reference template. The primers were designed by the conservative regions of the two pathogens, and the specific primers were screened according to the target bands and the dissolution curve and Ct value
of fluorescence quantitative PCR. At the same time, the extraction method of DNA kit of microbial in soil was improved, and the
detection systems of PCR and qPCR were established.【Result】A pair of specific primers PN3 and JBR for F. oxysporum and
P. parasitica var. nicotianae were screened respectively, and the template concentration ranged from 1×102
to 1×10-4 ng,
with good linear relationship. The number of two pathogens in diseased soil was higher than that in healthy soil. The method of
combination of soil drying, grinding and eluting for twice was helpful to extract the DNA of pathogens in the soil. A qPCR detection
method with the products obtained by ordinary PCR pre-amplification reaction as a template was established.【Conclusion】The
combination of PCR and qPCR can quickly distinguish and detect F. oxysporum and P. parasitica var. nicotianae, which plays an
important role in real-time monitoring and early warning of fusarium wilt and tobacco black shank. |
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