Method for identifying wheat leaf rust single sorus by utilizing nested PCR (polymerase chain reaction)

Abstract

The invention relates to a method for identifying a wheat leaf rust single sorus by utilizing nested PCR (polymerase chain reaction). The method comprises the following steps: designing and synthesizing a primer, extracting DNA (deoxyribonucleic acid) of the single sorus, performing amplification and separation on target genes, performing separation and identification on an amplified final product, then performing sequencing, comparing in a Genbank and determining the wheat leaf rust single sorus. The method for identifying the wheat leaf rust single sorus by utilizing the nested PCR is simple to operate and rapid and accurate in identification result, and can be applied into actual production.

Description

technical field [0001] The invention relates to an identification method, in particular to a method for identifying single spore piles of wheat leaf rust by nest PCR. Background technique [0002] Wheat rust, commonly known as "jaundice", is a type of fungal disease caused by rust fungi. During the growth and development period, the damaged wheat mainly manifests on the leaves or stalks, and bright yellow or reddish brown powder blister-like lesions appear. When observed under a microscope, it can be seen that there are many single-cell uredospores in the lesions, which are called uredospore piles. They are produced by mycelium that invades the wheat plant. In the whole life cycle of wheat rust, in addition to producing uredospores, it also produces other types of spores, the most can produce 5 kinds of spores, but the uredospores still cause serious damage. The uredospores attack and spread very quickly, and it only takes 8 to 12 days from invading new plants to producin...

AI Technical Summary

Problems solved by technology

Mechanical and physical wall breaking often requires certain equipment and technical conditions to achieve the purpose of wall breaking, such as ultrasonic crushers, etc., which cannot be fully equipped by grassroots plant protection institutions in my country at present

Chemical dissolution and wall breaking require the addition of chemical substances to dissolve the spore wall in the early stage. Most of the added substances are ethyl acetate, etc. After the wall breaking is completed, the DNA in it needs to be recovered and purified to remove chemical substances that may affect the next PCR reaction. , and the degree of completion of the removal will greatly affect the success rate of the next step

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