A method for amplifying the 16s rRNA gene of plant endophytes and its 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 invention relates to the technical field of gene amplification, in particular to a 16S rRNA gene amplification method and application of endophytic bacteria in plants. Background technique [0002] Plant endophytes are an important part of the plant micro-ecosystem, and they have formed an interdependent relationship with plants during the long-term co-evolution process. Plant endophytes can produce active substances as biological 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 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. However, even related research using high-throughput seq...

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