Method for detecting diversity of large aquatic plants based on environmental DNA technology

Abstract

The invention discloses a method for detecting diversity of large aquatic plants based on an environmental DNA technology. The method for detecting the diversity of the large aquatic plants in a waterarea to be detected comprises the following steps: 1) collecting a surface sediment sample of the water area to be detected; 2) extracting environmental DNA (eDNA) in the sediment sample; 3) amplifying the eDNA sample by using an Rbcl primer to obtain a PCR product, and recovering and purifying the product to obtain a purified PCR product; and 3) sequentially performing database building, sequencing, clustering and annotation on the purified PCR product to determine the species composition and / or the species relative abundance of the large aquatic plants in the water area to be detected, so as to realize the detection of the diversity of the large aquatic plants in the water area to be detected. The method disclosed by the invention has the following advantages: the sampling is simple andfast; the detection is efficient, the precision is high, and the detection result is accurate; and the large aquatic plants do not need to be collected, so that the aquatic plant population is protected, and particularly the protection of rare and endangered species is realized.

Description

technical field [0001] The invention relates to the field of molecular ecology, in particular to sample collection of water environment DNA, DNA extraction, primer amplification and species identification of macrobarcode sequences, and specifically refers to a method for detecting the diversity of large aquatic plants based on environmental DNA technology. Background technique [0002] Macrophytes refer to all aquatic plant groups that are physiologically attached to the water environment except microalgae. According to different life types, macrophytes in river habitats can be divided into aquatic plants (the entire life cycle is completed in water), amphibians (the life cycle can be completed in both water and shore wetlands) and mesophytes ( The entire life history is basically completed in the wetland on the shore). As an important part of the aquatic ecosystem, large aquatic plants are the basis for the health of the aquatic ecosystem and play a key role in maintaining...

AI Technical Summary

Problems solved by technology

The traditional assessment of the diversity of large aquatic plants mainly relies on the sample survey method, but the sample survey method is time-consuming, laborious and generally has the following errors: 1. Neglect of errors

It is easy to ignore the species that are rare in number, small in size and secretive in growth, resulting in incomplete investigation results

2. Identification error

Many plant identifications rely on flowers and fruits as key features, and species belonging to the same species or genus have extremely similar appearances, and not all plants exhibit flower and fruit characteristics during the investigation period, these factors easily lead to identification errors and test results Deviation

3. The estimated abundance is not accurate

Environmental DNA technology started late in the study of aquatic plant diversity. The selection of general primers, the screening of sampling sample types, and the validity and reliability evaluation of annotation results have not been scientifically verified and evaluated in large-scale aquatic plant diversity research.

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