PCR primer and quantitative detection method for rapidly detecting strawberry colletotrichum gloeosporioides

Abstract

The invention relates to a PCR primer and quantitative detection method for rapidly detecting strawberry colletotrichum gloeosporioides. The PCR primer is designed according to a cutinase gene consensus sequence of colletotrichum gloeosporioides physiological races and comprises two primers Cfcut1 and Cfcut2 and an amplification product 115bp; by utilizing the PCR primer and the quantitative detection method, fungal genome DNA or field sample DNA can be subjected to PCR detection without separation and culture of pathogenic bacteria, the amplification efficiency is high, the specificity is good, the amplification efficiency of the primer is 99.47%, and colletotrichum gloeosporioides compound species causing strawberry anthracnose can be rapidly detected and identified; and through real-time fluorescent quantitative PCR, absolute quantitative detection can be carried out on the biomass of the colletotrichum gloeosporioides in a sample, the occurrence period and the prevalence intensityof the anthracnose are predicted timely and accurately, the anthracnose is prevented and treated and even avoided, negative influences caused by use of pesticide can be reduced, and a basis is provided for comprehensive prevention and treatment of the anthracnose.

Description

technical field [0001] The invention belongs to the field of detection of plant pathogens, in particular to a PCR primer and a quantitative detection method for rapid detection of Gleosporium fragariae anthracnose. Background technique [0002] Strawberry belongs to Rosaceae, Fragaria genus, and is a perennial herb plant. It is one of the traditional excellent fruits in my country. As a characteristic and advantageous industry among horticultural crops in Shanghai, strawberry has very significant economic benefits. The suburban area is maintained at about 40,000 mu all year round. However, strawberry diseases have always restricted the development of strawberry industry. Strawberry anthracnose, as one of the main diseases in strawberry production, has an increasing trend in recent years. [0003] "Strawberry anthracnose" refers to all strawberry diseases caused by fungi of the genus Anthracnose. The reported pathogens of strawberry anthracnose mainly include Colletotrichum g...

AI Technical Summary

Problems solved by technology

Therefore, seedling quarantine is essential in the process of strawberry planting. However, due to the characteristics of latent infection of strawberry anthracnose, the infected seedlings do not show symptoms, which brings difficulties to detection.

[0005] The traditional identification method of anthrax requires the isolation and cultivation of pathogenic bacteria, and further judgment based on its culture characteristics and micro-morphological structure, which is time-consuming and labor-intensive; and, based on the marker genes ITS, β-tubulin, ACT, CAL, CHS, GAPDH The PCR-specific detection such as ect. has the characteristics of high information error rate and the inability to distinguish the relationship within the species. Although the phylogenetic analysis based on this multi-gene joint tree building is recognized as the standard operation for the identification of anthrax bacteria, due to its complexity and consumption It takes a long time and cannot be applied to rapid detection in the field

[0006] At present, the most important means of prevention and control of the disease is still the spraying of chemical pesticides. With the increase of the amount of pesticides, a series of problems such as the decline of fruit quality, environmental pollution, and the emergence of pathogen resistance have appeared.

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