Plant endophyte 16S rRNA gene amplification method and application

Abstract

The invention discloses a plant endophyte 16S rRNA gene amplification method and application. The amplification method includes the following steps of plant pretreating; plant-sample total DNA extracting; sample 16S rRNA gene amplifying, wherein sample 16S rRNA gene amplifying comprises 16S rRNA gene total-length emulsion PCR amplifying and 16S rRNA gene hypervariable-region / conserved-region amplifying sub-amplifying; high-throughput sequencing and biological information analyzing based on a Illumina platform, wherein high-throughput sequencing and biological information analyzing based on the Illumina platform comprises plant endophyte 16S rRNA gene amplicon purifying and recycling, amplicon sequencing library establishing, Illumina HiSeq sequencing and sequencing data bioinformatics analysis. The gene amplification method is used for high-throughput-sequencing plant disease detection and phytophagous animal enteric microorganism detection. Sequencing analysis of plant endophytic bacteria is carried out with the high-throughput sequencing technology, the data size is larger, the detection result is more complete, pollution of plant hosts is reduced to the maximum degree, the result multiformity is higher, and the cost is low.

Description

Technical field [0001] The present invention relates to the technical field of gene amplification, in particular to a method and application of 16S rRNA gene amplification of plant endophytes. Background technique [0002] Plant endophytes are an important part of the plant micro-ecosystem, and they have formed an interdependent relationship with plants in the process of long-term co-evolution. Plant endophytes can produce active substances as bio-textile resources, carriers of exogenous genes and sources of new drugs, and have great application prospects in the fields of agriculture, medicine and health. Although after years of research, the current research on plant endophytic bacteria is still in its infancy. The investigation of the distribution and abundance of plant endophytic bacteria in plants still relies on traditional methods such as separation and purification using DGGE with lower flux. Even if high-throughput sequencing is used for related research, because of the ...

AI Technical Summary

Problems solved by technology

Although after years of research, the research on plant endophytic bacteria is still in its infancy. The investigation of the distribution and abundance of plant endophytic bacteria in plants still relies on traditional methods such as separation and purification using low-throughput DGGE and other methods. , even the relevant research using high-throughput sequencing is still at the level of basic research because of the interference of the plastids and mitochondria of the plant host itself, resulting in the research of plant endophytic bacteria

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