Insect geographical tracing method based on intestinal microorganisms

A technology of intestinal microorganisms and microorganisms, applied in the field of insect geographic traceability and high-throughput sequencing, can solve problems such as difficult to speculate on the source of invasion

Pending Publication Date: 2021-04-30
检科智创(北京)生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] At present, the research technologies used in geographical traceability mainly include DNA barcode technology, restriction fragment length polymorphism (RFLP), single nucleotide polymorphism (single nucleotide polymorphism, SNP), microsatellite Although these technologies can break through the morphological method and provide powerful technical means for the taxonomic status, invasion source, and dispersal route of invasive insects, these technologies all study the genetic evolution of insects through differences at the gene level, thus inferring Its origin; many insects have genetically complex groups such as hybrids, cryptic species, or compound species, and it is difficult to deduce the origin of their invasion through genetic changes

Method used

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  • Insect geographical tracing method based on intestinal microorganisms
  • Insect geographical tracing method based on intestinal microorganisms
  • Insect geographical tracing method based on intestinal microorganisms

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1. Method principle

[0034] According to the close relationship between the diversity of gut microbes and feeding and geographical environment, we collected harmful insects locally distributed across the country, extracted genomes, performed high-throughput sequencing of 16S hypervariable regions, and clustered gut microbes from different geographically and different insects According to the analysis, different insects can be distinguished according to the difference of microbial phylum level, and the same insects from different geographical sources can be distinguished according to the difference of microbial genus level.

[0035] 2. Instruments and equipment

[0036] PCR instrument, electrophoresis instrument, gel imaging system, high-speed refrigerated centrifuge, sterilizer, desktop small centrifuge, ultra-low temperature refrigerator, conventional refrigerator, vortex shaker, micro sample injector.

[0037] 3. Method flow

[0038] (1) Preparation of insect sampl...

Embodiment 2

[0055] The sample insects were collected from areas where the potato beetle is known to be colonized, including Wenquan County, Chabuchar County, Urumqi City, Xinyuan County, Tacheng City, Jimunai County in the Xinjiang Uygur Autonomous Region, and Suifenhe City in Heilongjiang Province. The samples in Tacheng City were divided into varieties collected on potato crops and eggplant crops, and the samples in Xinyuan County were collected in two plots. The collection information is shown in Table 2. Referring to the method of Example 1, the collected potato beetle was subjected to pretreatment, genomic DNA extraction, and two PCR amplifications, and then the PCR product was purified, the DNA concentration was determined, and the purified product was subjected to high-throughput sequencing.

[0056] Table 2 Geographic collection information table of potato beetle samples

[0057]

[0058]

[0059] The intestinal microbial diversity of potato beetles collected from existing p...

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Abstract

The invention relates to an insect geographical tracing method based on intestinal microorganisms. The metagenome sampling and sequencing technology is utilized to perform high-throughput sequencing on insect genome 16SDNA hypervariable regions, cluster analysis is performed on intestinal microorganisms of insects from different geographical sources, and it is judged that different insects can be distinguished according to the difference of microbial gate levels. The same kind of insects with different geographical sources can be distinguished according to the difference of microbial genus levels, so that when possible harmful insects are investigated or monitored, the invasion source of the insects can be judged by utilizing the diversity of intestinal microorganisms of the insects, and the insects with different geographical sources can be accurately distinguished.

Description

technical field [0001] The invention relates to the technical field of insect geographic origin tracing and high-throughput sequencing, in particular to a method for insect geographic origin tracing based on intestinal microorganisms. Background technique [0002] At present, the research technologies used in geographical traceability mainly include DNA barcode technology, restriction fragment length polymorphism (RFLP), single nucleotide polymorphism (single nucleotide polymorphism, SNP), microsatellite Although these technologies can break through the morphological method and provide powerful technical means for the taxonomic status, invasion source, and dispersal route of invasive insects, these technologies all study the genetic evolution of insects through differences at the gene level, thus inferring Its source: Many insects have genetically complex groups such as hybrids, cryptic species, or compound species, and it is difficult to speculate on the source of their inv...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/6869C12Q1/10C12Q1/06C12Q1/04C12R1/145C12R1/38C12R1/01
CPCC12Q1/689C12Q1/6869
Inventor 师科
Owner 检科智创(北京)生物科技有限公司
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