Primer sequence for testing imazalil fastness frequency of fingerlike penicillium notatum and testing method

Abstract

The invention discloses a primer sequence which is used to detect the resistance frequency of Penicillium digitatum to imazalil, which comprises three groups of primer. A sense primer of the first group is equipped with a base sequence of SEQ ID No.1 in the sequence table, and an antisense primer of the first group is equipped with a base sequence of SEQ ID No.2 in the sequence table; a sense primer of the second group is equipped with a base sequence of SEQ ID No.3 in the sequence table, and an antisense primer of the second group is equipped with a base sequence of SEQ ID No.4 in the sequence table; a sense primer of the third group is equipped with a base sequence of SEQ ID No.5 in the sequence table, and an antisense primer of the third group is equipped with a base sequence of SEQ ID No.4 in the sequence table. The primer sequence of the invention has very high specificity and lays the foundation of the entire detection system.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a primer sequence and a detection method for detecting the frequency of resistance of Penicillium digitatum to imazalil. Background technique [0002] my country is the third largest country in the world for citrus production, with an output of about 15 million metric tons in 2004, second only to Brazil and the United States. Citrus green mold caused by Penicillium digitatum is the most important disease during postharvest storage and marketing of citrus. The fungus source in the packaging storage warehouse is the main source of primary infection causing rot during storage. If no chemical treatment is used, the disease can cause 20-30% of post-harvest citrus to rot in a normal year. [0003] The use of fungicides is one of the most important measures for controlling postharvest rot in citrus. In the early 1990s, my country began to widely use the fungicide imazalil to treat pos...

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Problems solved by technology

[0006] Based on this molecular mechanism of resistance, there have been reports in the world using traditional PCR technology to detect resistance (Hamamoto H, Hasegawa K, Nakaune R, et al. PCR-based detection of sterol demethylation inhibitor-resistant strains of Penicillium digitatum[J]. PestManagement Science, 2001, 57: 839-843.), but this method can only detect the resistance of one bacterial strain at a time. If the proportion of resistant individuals in the bacterial population needs to be determined, a large number of bacterial strains need to be detected, which is more time-consuming It is laborious, and this method can only detect the proportion of resistant strains in the isolated strains, and cannot reflect the proportion of the total individual amount of resistant strains in the total population of pathogenic bacteria in the storage warehouse, that is, the frequency of resistance. Therefore, in most cases It is difficult to effectively carry out early warning and detection of bacterial resistance

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