Molecular detection method for Fusarium circinatum

Abstract

The invention discloses a molecular detecting method of pine grease ulcer germ, which is characterized by the following: utilizing primer designing software Prime 5.1 to design a couple of general prime G1 / G2 and astopic prime S1 / S2 on the fusarium IGS segment; adopting pine germ sample DNA extract as form; proceeding the first-turn PCR augmenting reaction of general prime; diluting the PCR augmented product by 100 times; fetching 1 mul as form to proceed the second-turn astopic prime PCR augmenting reaction; proceeding electrophoretic detection for augmented product; affirming the detected germ as pine grease ulcer germ if the DNA astopic band with 873bp exists. The invention needs only one working day to finish detect, whose density of reaches minimum quantity germ DNA is 5*10-3pg / mul and conidiospore is 10units / 100 mul.

Description

technical field [0001] The invention relates to a method for detecting pine resin canker (Fusarium circinatum Nirenberg & O'Donnell, whose sexual stage is Gibberellacircinata Nirenberg & O'Donnell), which is suitable for use in departments such as animal and plant quarantine, forestry production, and plant protection. In particular, a simple and rapid molecular detection method belongs to the field of biology. Background technique [0002] Pine resin canker is one of the most serious diseases of pine trees in the world today. In 1946, pine resin canker disease was first discovered on North Carolina dwarf pine (P. virginiana) in the southeastern United States. It mainly caused symptoms such as sap and ulcers on the trunk and branches. The disease was first discovered in California in 1986, and then the local distribution continued to expand, and the number of harmful hosts also increased, including Pinus, Douglas-fir and other tree species. In addition, Japan in 1989, Mexic...

AI Technical Summary

Problems solved by technology

However, looking at the above methods, some experiments are not repeatable, some operations are cumbersome, and the detection cost is expensive. They all fail to solve the problem of directly detecting pine resin canker bacteria from samples, and it is difficult to directly apply them at ports.