High-throughput-sequencing and host-background-interference-free method for amplifying ITS genes of endophytic fungi of vigna unguiculata

Abstract

The invention discloses a high-throughput-sequencing and host-background-interference-free method for amplifying ITS genes of endophytic fungi of vigna unguiculata. According to the method, a modified oligonucleotide primer combination is adopted, the amplification of host-derived ITS genes of the vigna unguiculata is removed through a multiple PCR amplification method, meanwhile, the amplification of the ITS genes of the endophytic fungi of the vigna unguiculata is not affected, finally, the variety and abundance of the endophytic fungi (including pathogenic microorganisms) of the vigna unguiculata can be surveyed and understood comprehensively through a high-throughput sequencing method, and thus, a reliable new way is provided for identifying the endophytic fungi of the vigna unguiculata. Compared with the traditional methods which comprise the step of carrying out identification after separation and purification, the method has the advantage that the existing technical limitations are overcome. By the method, environmental pollution resulting from complicated operating processes can be reduced, the required sample size is small, the test operation is simple and easy, and the level of research on fungus diseases of the vigna unguiculata can be greatly increased.

Description

technical field [0001] The invention relates to a technology for amplifying nucleic acid target sequence fragments of endophytic fungi in cowpea, in particular to an ITS gene amplification method for endophytic fungi in cowpea without host background interference for high-throughput sequencing. Background technique [0002] Vegetable fungal diseases account for more than 80% of the disease types, and the diseases caused by fungal infection of vegetables are quite serious. The fungus can invade the host through the rhizosphere soil or spores under wind and rain conditions from the epidermis, and can also invade through wounds, etc. The pathogenic fungus remains in the diseased residue, seeds, soil, etc. to survive the winter, and can harm the host under suitable conditions. [0003] As one of the most important legume crops in the world, cowpea is widely cultivated in tropical and subtropical regions and some temperate regions, the Mediterranean Basin and the southern United ...

AI Technical Summary

Problems solved by technology

However, due to the high homology of eukaryotic ITS genes, even if high-throughput sequencing is used for related research, the research on plant endophytic fungi is at the basic research level due to the interference of the plant host's own genome

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