Monitoring method for natural reproduction of fish based on environmental DNA technology

A technology of natural reproduction and technology, applied in the field of molecular ecology, can solve the problems of high operation difficulty, high labor intensity, and different hydrological conditions, and achieve the effect of simplified operation steps and high detection sensitivity

Inactive Publication Date: 2017-08-29
CHINESE STURGEON RES INST CHINA THREE GOR
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although the above method can monitor the spawning behavior of domestic fish and locate the range of spawning grounds, due to the difference in flow and hydrological conditions of different rivers, there is a large uncertaint

Method used

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  • Monitoring method for natural reproduction of fish based on environmental DNA technology
  • Monitoring method for natural reproduction of fish based on environmental DNA technology
  • Monitoring method for natural reproduction of fish based on environmental DNA technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The natural reproduction of domestic fish during the first operation of the Three Gorges Reservoir in 2016 (June 8-12, 2016). 1. Sample collection

[0034] According to many years of traditional survey results, six sections were set up to collect water samples within the 36-kilometer section from Yichang Miaozui Wharf to Houjiangtuo. Take 2L of mixed water samples from water layers of different depths and store them in an incubator at 4°C. The specific geographical locations of the sampling points are shown in Table 1. 2. DNA extraction

[0035] Vacuum filter the above water sample through a fat-soluble filter membrane with a pore size of 0.22 μm within 24 hours. According to multiple pre-experimental comparisons, one filter membrane should be used for each liter of water, so put the filtered two filter membranes into Store in a 50ml centrifuge tube at -20°C for later use.

[0036] Using a professional water sample DNA extraction kit the Water DNA Kit (Omega) was ...

Embodiment 2

[0058] Embodiment 2: The natural breeding situation of domestic fish when the Three Gorges Reservoir was dispatched for the second time in 2016 (June 19th-22nd, 2016), the method steps are the same as in Example 1, and Table 3 is 6 sections and 18 sampling points are continuous For the DNA copy number of domestic fish obtained by ddPCR quantitative analysis in 4 days, the daily average total concentration can be found in image 3 , the concentration of each section see Figure 4 .

[0059] Table 3. DNA copy numbers of domestic fish detected during the second dispatch

[0060]

[0061] According to the experimental results of ddPCR, the copy number of the target species contained in each sample is known, and the daily average total DNA concentration of domestic fish eDNA in 6 sections of the monitoring period is compared. It can be known that the domestic fish environment on the first day of the second ecological dispatch DNA content is maximum. Comparing the DNA concentr...

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Abstract

The invention relates to a monitoring method for natural reproduction of fish based on environmental DNA technology. The method comprises the following steps: acquisition of a water sample; DNA extraction; ddPCR amplification of a target fragment; and concentration analysis. The monitoring method provided by the invention is based on environmental DNA identification technology, does not depend on traditional fishing technology for roe and fries and traditional anatomical technology for oophagous fish and can determine the oviposition behavior of a target species and the position of an oviposition site only through reasonable design of a sampling point, acquisition of a water sample and concentration analysis of the target species. The method maximally protects fish resources and avoids inconvenience caused by restrictive factors like long time for traditional fishing and difficulty in fishing of a part of fish fingerlings.

Description

technical field [0001] The invention relates to the technical field of molecular ecology, in particular to the fish species concentration analysis of water body environmental DNA samples, in particular to a method for monitoring the natural reproduction of fish based on environmental DNA technology. Background technique [0002] Due to the construction of water conservancy projects, environmental pollution and changes, overfishing and other reasons, the number and diversity of fish populations in natural water bodies such as the Yangtze River Basin have been decreasing year by year, and the construction of dams has blocked the passage of migratory fish. The establishment of the project cannot be traced back to the original spawning ground for breeding, and a new spawning ground is formed under Gezhouba. For example, the establishment of the reservoir area has slowly evolved the fish community structure, which was dominated by flowing water fish, into a structure composed mos...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6888C12Q2563/159C12Q2545/114C12Q2563/107
Inventor 李莎刘雪清姜伟郜星晨黄涛
Owner CHINESE STURGEON RES INST CHINA THREE GOR
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