Method for identifying and detecting porphyra red rot pythium pathogens

Abstract

The invention discloses a method for identifying and detecting porphyra red rot pythium pathogens, and belongs to the technical field of algae cultivation disease detection, identification and control. According to the method, specific primers are designed on the basis of genomic difference sequences of the porphyra red rot pathogens P.porphyrae and P.chondricola, and a single PCR detection method and a double PCR detection method for the two pythium pathogens are established. By the method, the P.porphyrae and the P.chondricola as well as the possibility of coexistence of the P.porphyrae and the P.chondricola can be identified and detected. The lower limit of single PCR primer P-for / P-rev detection is 103 copies, and the lower limit of C-for / P-rev primer detection is 102 copies. The lower limit of double PCR detection is 104 copies. According to the method, a specific and sensitive porphyra red rot pythium pathogen identification method is established, the P.porphyrae and the P.chondricola can be identified without sequencing and comparison, and a technical support is provided for prevention and control of red rot in porphyra cultivation in China.

Description

technical field [0001] The invention belongs to the technical field of detection, identification and prevention of algae cultivation diseases, and in particular relates to an identification and detection method of red rot pathogen in laver cultivation—Pythium pathogen. Background technique [0002] Red rot is the main disease of laver during the cultivation period, and it often occurs in sea areas with high water temperature, low salinity, poor water exchange, and insufficient drying time. The onset of red rot is rapid, and within a week after the outbreak, the laver net curtain can become empty, resulting in a 30-50% reduction in production, and it can reach more than 60% in some sea areas. In the early stage of the disease, red circular lesions with a diameter of about 2 mm can be seen on the leaves. Within 2 days, the diameter of the lesions can expand to 4-5 mm. At this time, the lesions are red on the edge and yellow-green in the middle. Enlargement, several lesions th...

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Problems solved by technology

In 2016, Wu Shangzhe established a fluorescent quantitative PCR detection method for the ITS sequence of P.porphyrae, which can shorten the detection time and has high sensitivity, but the error of the actual sample is large

[0005] Among the two Pythium pathogens of Porphyra, P.porphyrae and P.chondricola are different in pathogenicity; however, they are completely different in mycelial morphology, reproductive mode, infection characteristics and internal transcribed spacer (ITS) sequence. The same, can not be distinguished by the above characteristics, can only be distinguished by the difference of 4 bases in the gene sequence of cytochrome c oxidase subunit 1 and 2 (cytochrome c oxidase subunit I and II, cox 1 and cox 2)

At present, none of the detection methods for the pathogen of laver red rot can distinguish between P.porphyrae and P.chondricola

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