Fluorescent quantitative PCR detection method for detecting colonization amount of tobacco anthracnose

Abstract

The invention relates to a fluorescent quantitative PCR detection method for detecting colonization amount of tobacco anthracnose. The method comprises the following steps: (1) preparation of a fluorescent quantitative nucleic acid amplification reaction system: a forward LNA primer, a reverse LNA primer, a template DNA and 2x fluorescent quantitative PCR reaction mixed liquor are added into a reaction tube, and 25[mu]l of ddH2O is added into the reaction system with uniformly mixing; (2) fluorescent quantitative nucleic acid amplification: the mixed liquor obtained in the step (1) is placed in a fluorescent quantitative detector for carrying out a reaction; (3) after the reaction ends, the cycle threshold C(t) of the template DNA is compared with a standard curve, and concentration of 18S ribosome RNA fragment copies in the template DNA is obtained. The method has the advantages of short detection time, high sensitivity, simple operation process, standardized operation, and high-throughput detection.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a fluorescent quantitative PCR detection method for detecting the colonization amount of tobacco anthracnose, which is suitable for the quantitative detection of tobacco anthracnose bacteria in various susceptible tobacco strains. Background technique [0002] After tobacco anthrax was first reported by Brazil in 1922, it was also discovered in Germany, Japan, the United States, China, Australia, India, North Korea and Africa. The disease can occur in all growth stages of tobacco, but it is common and serious at the seedling stage. The leaves of seedlings are densely covered with diseased spots, and the whole tobacco seedlings are often destroyed when the disease is severe. Generally, although the seedlings are not destroyed when the disease occurs, the seedlings have poor growth potential, and they can continue to cause damage after transplanting to the field, resulting in great...

AI Technical Summary

Problems solved by technology

These methods are time-consuming and error-prone

For example, pear ringworm and pear anthracnose have similar symptoms on fruits and leaves, and it is difficult to distinguish them by traditional monitoring methods.

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