Multiple PCR detection kit for three bacterial seed-borne diseases and special primer for kit as well as multiple PCR detection method

Abstract

The invention discloses a multiple PCR detection kit for three bacterial seed-borne diseases and a special primer for the kit as well as a multiple PCR detection method. The invention provides special primers aiming at the canker pathogens of tomato, the bacterial leaf spot pathogens of the tomato and the bacterial fruit spot pathogens of melons; the target product fragment of the special primer for detecting the canker pathogens of the tomato is 1086bp; the target product fragment of the special primer for detecting the bacterial leaf spot pathogens of the tomato is 491bp; and the target product fragment of the special primer AT for detecting the bacterial fruit spot pathogens of the melons is 290bp. The invention also provides a multiple PCR optimal reaction system for the three pathogens and reaction conditions. The multiple PCR detection kit is capable of quickly and simultaneously detecting the canker pathogens of the tomato, the bacterial fruit spot pathogens of melons and the bacterial leaf spot pathogens of the tomato without cloning, sequencing and sequence alignment, and is high in specificity, quick and accurate; and as a result, the detection cost and the detection time are saved, and the limitation of the single reaction is solved.

Description

technical field [0001] The present invention relates to the technical field of molecular biology detection of three kinds of plant pathogens, in particular to Clavibacter michiganensis subsp.michiganensis, tomato bacterial leaf spot (pseudomonas syringae pv.tomato) and melon bacterial fruit spot ( Acidovorax citrulli) multiplex molecular biology detection method. Background technique [0002] my country is the fourth largest importer and fifth largest exporter of agricultural products. However, with the further expansion of foreign trade, the transportation of plants and their products has become increasingly frequent, especially the most dangerous seeds. This has become a very important and urgent problem that endangers my country's biodiversity, ecological environment and national economy. The number and variety of alien invasive organisms are large. Already 529 species of alien organisms have invaded 34 provincial-level administrative regions in my country, leading to spe...

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Problems solved by technology

In 2000, bacterial fruit spot disease of thick-skinned muskmelons occurred on a large scale in Bayannaoer League, Inner Mongolia, with an average yield reduction of 46%, and the commercial melon rate was only 1 / 3; in the winter of 2002, melon bacterial fruit spot disease The seedling destruction rate that causes is also up to 30%~80%, causes serious economic loss to melon grower

However, the detection methods described in the above-mentioned patents are only aimed at the detection of Clavibacter michiganensis subsp. michiganensis and Acidovorax citrulli respectively, and cannot detect Clavibacter michiganensis subsp. michiganensis at the same time. , bacterial leaf spot of tomato (Pseudomonas syringae pv.

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