Method for detecting phytophthora parasitica in soil

Abstract

The invention belongs to the field of plant disease and pest quarantine, and provides a method for detecting phytophthora parasitica in soil. According to the method for DNA extraction and molecular detection of the phytophthora parasitica in the soil, the conventional isolating culture method is eliminated, and the detection period is greatly shortened; compared with a common polymerase chain reaction (PCR) detection method, the method has the advantages that: a subsequent treatment process is not needed, and a result can be monitored in real time; primers are designed on the basis of a changeful area of pathogen ITS, and corresponding bands are not produced when similar species are amplified; the method is high in specificity and suitable for fluorescent quantitative PCR conditions; because 2 percent of polyvinyl pyrrolidone (PVP) is added into the total DNA extraction buffer solution of the soil, the pollution of partial impurities in the soil is effectively removed, the influence of the impurities on the PCR effect is reduced, and the detection sensitivity of fluorescent quantitative PCR is improved; meanwhile, the fluorescent quantitative PCR system and the reaction conditions are obtained according to the size study conditions of the designed primers, so the detection sensitivity is high and can reach 2fg / mu l.

Description

technical field [0001] The invention belongs to the field of plant disease and insect pest quarantine, and in particular relates to a method for detecting tobacco blackleg bacteria in soil. Background technique [0002] Tobacco is an annual herbaceous plant belonging to the Solanaceae (Solanaceae) genus Nicotiana (Nicotiana), widely distributed in the world, and there are 66 species of Nicotiana found so far. In 2010, the tax and profit of the national tobacco industry exceeded 600 billion yuan. Tobacco black shank is one of the most destructive infectious diseases in tobacco production. In my country, the annual incidence rate is maintained at 10-15%, and the diseased area is about 76372hm2. The annual loss of output is about 2869.26Kg, and the economic loss is up to 1.23 billion yuan or more. The current research shows that the amount of black shank pathogenic bacteria in soil is closely related to the occurrence of tobacco black shank. Therefore, detecting the number of...

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Problems solved by technology

[0004] The invention provides a method for detecting tobacco black shank bacteria in soil, aiming at solving the problem that the currently commonly used detection methods for black shank bacteria in soil are mainly through cell culture and common PCR method, and these two methods are accurate. Inefficient, time-consuming and poorly reproducible

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