Conjoint analysis method for estimating DNA abundance of fishes based on environment DNA technology

Abstract

The invention relates to a conjoint analysis method for estimating the DNA abundance of fishes based on an environment DNA technology. The conjoint analysis method includes the steps that the total DNA abundance of the fishes is acquired through timed and quantified PCR; the species composition proportion of the fishes is acquired through 454 GS-FLX Titanium sequencing analysis; conjoint analysis of target species is performed through sequencing data and timed and quantified PCR data; a relative distribution diagram of mitochondrial DNA of the fishes in an environment is drawn through sufer 8.0 software. Based on the environment DNA identification technology, weather species exist and how many species exist can be analyzed just by collecting water samples without depending on fish species capturing, a sampling method is simple, and fish resources can be protected to the maximum degree; meanwhile, for some species which are rarely distributed and difficult to capture, the method is still more effective than traditional fishing investigation, the method can effectively solve the problem that effective species specificity primers are difficult to obtain when multiple closely related species exist in samples, and NDA abundance information of all species can be known just by conducting experiment once respectively through the two technologies.

Description

technical field [0001] The present invention relates to the technical field of fish quantity assessment in molecular ecology, in particular to the identification, differentiation and quantitative analysis of closely related species of fish by samples of water body environmental DNA, and specifically refers to a method based on environmental DNA technology to analyze the abundance of fish DNA. Conjoint analysis of estimates. Background technique [0002] The current domestic methods for fish count mainly include marking recapture method and acoustic detection method. The marking and re-catching method is to carry out statistics on the number of fish species by marking the fish in the water body and then fishing. The realization of this method is mainly to catch fish by various markings and fishing tools (such as various nets, fishing tackle, electric catch, etc.). This method has the following problems: 1. The selection and placement of the marks requires a lot of work, whi...

AI Technical Summary

Problems solved by technology

There are the following problems in this method: 1. The selection and placement of markers requires a lot of work, which consumes manpower and material resources.

2. High requirements for recapture intensity

A large amount of fishing work is required, and a large enough number of recaptures is required, and the cost is large and it is difficult to ensure accuracy

3. This method needs to capture live bodies, it is difficult to ensure the survival of the captured fish, and it will cause certain damage to fish resources

4. For the monitoring and investigation of rare species and exotic species, due to their scarcity, they cannot be effectively marked and recaptured, and it is difficult to achieve good results, and the investigation results are more likely to be biased

However, when we are studying some important native species, such as certain species of carps, due to the existence of a large number of closely related species, the amplification of mitochondrial genes in mixed environmental DNA samples containing DNA from multiple species When increasing, it is often difficult to obtain effective species-specific primers. This problem makes it difficult for us to obtain the DNA abundance information of the target species by direct quantitative PCR. This problem has not been effectively resolved.

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