Method for quantitatively detecting colletotrichum gloeosporioides by utilizing real-time fluorescent PCR (Polymerase Chain Reaction)

Abstract

The invention provides a method for quantitatively detecting colletotrichum gloeosporioides by utilizing real-time fluorescent PCR (Polymerase Chain Reaction). A real-time fluorescent quantitative PCR detection method of colletotrichum gloeosporioides is established through primers designed by actin genes of colletotrichum gloeosporioides; the detection method disclosed by the invention is high in specificity and sensitivity, can detect colletotrichum gloeosporioides in an incubation period, and has important significance for detecting potential colletotrichum gloeosporioides infection; according to the detection method disclosed by the invention, the normalization relation between pathogenic bacterium DNA and plant DNA biomass is also considered, the growth conditions of colletotrichum gloeosporioides in different host plants can be accurately determined, the method can be used for screening and identifying disease-resistant varieties of arabidopsis thaliana and stylosanthes guieosporioides, and a basis is provided for early detection and timely prevention and treatment of colletotrichum gloeosporioides.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for quantitatively detecting anthracnose by real-time fluorescent PCR. Background technique [0002] Traditionally, the diagnosis of pathogenic bacteria infection relies on the isolation and culture of pathogenic bacteria, which is time-consuming and labor-intensive. The easiest way to detect phytopathogens is visual disease symptom assessment, but visible lesions usually occur late in the host-pathogen interaction, making early control of anthracnose difficult. [0003] With the development of the era of genomics, real-time PCR has developed into a method that can rapidly identify pathogens and diagnose diseases. For example, Hong Yantao et al. used RT-PCR to detect the relative expression of the Rhizoctonia graminearum actin gene in wheat to identify the resistance of different wheat varieties to pathogens [Hong Yantao, Zhang Zengyan. Detection of cereal grains by fluores...

AI Technical Summary

Problems solved by technology

However, compared with Rhizoctonia graminearum, the hosts that M. glyospora can infect are widely distributed, especially in tropical and subtropical crop regions, including strawberry, mango, pepper, tomato, cassava, pomegranate, stylo and Hevea brasiliensis, etc. , and there are obvious differences in the strain characteristics and infection process between Rhizoctonia graminearum and Anthracnose glyospora

Gloospora anthracnose exhibits a semi-biotrophic lifestyle, and its pathogen invasion process includes a biotrophic stage and a destructive necrotrophic stage. In the latent state for a long time, conventional methods can only be identified after the disease is manifested, and the detection sensitivity is low. 0.57ng / μL DNA, the sensitivity is not high enough, it is easy to miss the best time for prevention and treatment, causing serious economic losses

Moreover, the field hazard symptoms of anthracnose pathogens such as G. oxysporum and G. oxysporum are basically similar, and the existing identification methods are difficult to distinguish. Therefore, accurate detection of potential G. anthracnose is of great significance for reducing the spread of G. important meaning

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