CN115044663A - Method for rapidly and accurately evaluating diversity of ecological fishery resources on large water surface - Google Patents
Method for rapidly and accurately evaluating diversity of ecological fishery resources on large water surface Download PDFInfo
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- CN115044663A CN115044663A CN202210824479.5A CN202210824479A CN115044663A CN 115044663 A CN115044663 A CN 115044663A CN 202210824479 A CN202210824479 A CN 202210824479A CN 115044663 A CN115044663 A CN 115044663A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6869—Methods for sequencing
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- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06Q—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR ADMINISTRATIVE, COMMERCIAL, FINANCIAL, MANAGERIAL OR SUPERVISORY PURPOSES; SYSTEMS OR METHODS SPECIALLY ADAPTED FOR ADMINISTRATIVE, COMMERCIAL, FINANCIAL, MANAGERIAL OR SUPERVISORY PURPOSES, NOT OTHERWISE PROVIDED FOR
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- G06Q10/063—Operations research, analysis or management
- G06Q10/0639—Performance analysis of employees; Performance analysis of enterprise or organisation operations
- G06Q10/06393—Score-carding, benchmarking or key performance indicator [KPI] analysis
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- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06Q—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR ADMINISTRATIVE, COMMERCIAL, FINANCIAL, MANAGERIAL OR SUPERVISORY PURPOSES; SYSTEMS OR METHODS SPECIALLY ADAPTED FOR ADMINISTRATIVE, COMMERCIAL, FINANCIAL, MANAGERIAL OR SUPERVISORY PURPOSES, NOT OTHERWISE PROVIDED FOR
- G06Q50/00—Systems or methods specially adapted for specific business sectors, e.g. utilities or tourism
- G06Q50/02—Agriculture; Fishing; Mining
Abstract
The invention relates to the technical field of large-water-surface ecological fishery resource assessment, in particular to a method for rapidly and accurately assessing diversity of large-water-surface ecological fishery resources, which comprises the following steps: the method has the advantages that the ecological fishery resource diversity on the large water surface can be rapidly and accurately evaluated, and the problems of ecological destructiveness, high requirement on professional morphological knowledge of an inspector, large investigation workload and the like of the traditional fish monitoring method are effectively solved.
Description
Technical Field
The invention relates to the technical field of large-water-surface ecological fishery resource assessment, in particular to a method for rapidly and accurately assessing diversity of large-water-surface ecological fishery resources.
Background
The large water surface fishery plays an important role in the field of aquaculture in China, and the area and the yield of the large water surface fishery respectively account for 46% and 15.42% of fresh water fishery in China. However, the current large-water-surface fishery faces the problems that the ecological imbalance of reservoirs and lakes, even the blue algae outbreak and the like can be caused by the problems that the seedlings are thrown depending on experience and the effective ecological regulation is lacked. In the production process of the large-water-surface ecological fishery, whether reasonable seedling throwing or ecological regulation and control is carried out, the diversity of fishery resources needs to be evaluated quickly and accurately. At present, on-site fishing and morphological identification are mainly adopted for large-water-surface ecological fishery resource assessment, time and labor are wasted, accuracy is not high, and fishery resource diversity is difficult to assess rapidly and accurately.
Disclosure of Invention
The invention aims to solve the defects in the background art and provides a method for quickly and accurately evaluating the diversity of ecological fishery resources on a large water surface.
In order to achieve the above purposes, the technical scheme adopted by the invention is as follows: a method for rapidly and accurately evaluating diversity of ecological fishery resources on a large water surface comprises the following steps: sampling point position arrangement, sample collection, environmental DNA extraction and amplification, high-throughput sequencing, macro-barcode detection and fishery resource diversity analysis.
Preferably, more than 5 sampling samples in a typical water area of a reservoir/lake should be selected for sampling point location.
Preferably, typical water areas include areas of heterogeneity, reservoir inlets, outlets, near shore and near center regions.
Preferably, the sample collected includes a water sample and a sediment sample.
Preferably, the water in the collecting reservoir comprises upper water, the water taking amount is 0.5-1L, and the mixed cellulose ester membrane filtration membrane with the diameter of 0.45um is used for suction filtration on site or suction filtration in an laboratory within 24 hours after refrigeration.
Preferably, a mud sampler is used for collecting sediments in a water sampling place during sediment sample collection, and the sediments are respectively stored in a sterile sealable wide-mouth bottle after passing through large granular substances in the sunning and rolling filter tank, wherein the large granular substances comprise large stones and plant branches.
Preferably, the sampling device in sample collection needs to be cleaned before each use to avoid cross-contamination.
Preferably, in the environmental DNA extraction and amplification, lysis buffer solution and proteinase K are respectively added into sample tissues, and after water bath lysis at 56 ℃, a DNA extraction kit is used for extracting the DNA of a water sample or sediment sample; the NDA extraction kit comprises any one of a Power Water DNA extraction kit and a DNeasy Blood and Tissue DNA extraction kit; sample DNA is amplified by using primers designed from any one of the sequences of mitochondrial cytochrome b, 12S, 16S or COI of the species in the local water area.
Preferably, high throughput sequencing and macro barcode detection: and (3) freezing the amplified sequences by liquid nitrogen, quickly transporting the sequences to a biological company by adding dry ice for high-throughput sequencing and macro-barcode detection, and annotating the species information represented by each sequence.
Preferably, the fishery resource diversity analysis comprises the following steps: and analyzing the diversity of fishery resources according to the compared species information by using the relative abundance of species, Alpha diversity, Shannon-Wiener index, Simpson index and Chaol index.
Compared with the prior art, the invention has the following beneficial effects:
the method can quickly and accurately evaluate the diversity of the ecological fishery resources on the large water surface, and effectively solves the problems of ecological destructiveness, high requirement on professional morphological knowledge of an inspector, large investigation workload and the like of the traditional fish monitoring method.
Detailed Description
The following description is presented to disclose the invention so as to enable any person skilled in the art to practice the invention. The preferred embodiments in the following description are given by way of example only, and other obvious variations will occur to those skilled in the art.
A method for rapidly and accurately evaluating diversity of ecological fishery resources on a large water surface comprises the following steps:
s1, arranging sampling points: more than 5 samples are collected in a typical water area of a reservoir/lake. Typical waters include areas of heterogeneity, reservoir inlets, outlets, near shore and near center regions.
S2, sample collection: collecting a water sample and a sediment sample, wherein:
the water in the collecting reservoir comprises upper layer water, the water taking amount is 0.5-1L, and the filtration is carried out by using a mixed cellulose ester membrane filtration membrane with the diameter of 0.45um on site or in an experimental room after refrigeration for 24 hours.
In the sediment sample collection, a mud sampler is used for collecting sediments in a water sampling place, the sediments are respectively stored in a sterile sealable wide-mouth bottle after passing through large granular substances in the sunning and rolling filter tank, and the large granular substances comprise large stones and plant branches.
The sampling devices need to be cleaned to avoid cross-contamination prior to each use in the sample collection described above.
S3, environmental DNA extraction and amplification: adding lysis buffer solution and proteinase K into sample tissues respectively, and extracting the DNA of a water sample or sediment sample by using a DNA extraction kit after the sample tissues are lysed in a 56 ℃ water bath; the NDA extraction kit comprises any one of a Power Water DNA extraction kit and a DNeasy Blood and Tissue DNA extraction kit; sample DNA is amplified by using primers designed from any one of the sequences of mitochondrial cytochrome b, 12S, 16S or COI of the species in the local water area.
S4, high throughput sequencing and macro barcode detection: and (3) freezing the amplified sequences by liquid nitrogen, quickly transporting the sequences to a biological company by adding dry ice for high-throughput sequencing and macro-barcode detection, and annotating the species information represented by each sequence.
S5, analyzing the diversity of fishery resources: and analyzing the diversity of fishery resources according to the compared species information by using the relative abundance of species, Alpha diversity, Shannon-Wiener index, Simpson index and Chaol index.
The method can quickly and accurately evaluate the diversity of the ecological fishery resources on the large water surface, and effectively solves the problems that the traditional fish monitoring method has ecological destructiveness, high requirement on professional morphological knowledge of an inspector, large investigation workload and the like.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are merely illustrative of the principles of the invention, but that various changes and modifications may be made without departing from the spirit and scope of the invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (10)
1. A method for rapidly and accurately evaluating diversity of ecological fishery resources on a large water surface is characterized by comprising the following steps: the method comprises the following steps: sampling point position arrangement, sample collection, environmental DNA extraction and amplification, high-throughput sequencing, macro-barcode detection and fishery resource diversity analysis.
2. The method for rapidly and accurately evaluating the diversity of the ecological fishery resources on the large water surface according to claim 1, wherein the method comprises the following steps: more than 5 sampling samples in a typical water area of a reservoir/lake should be selected for sampling point location arrangement.
3. The method for rapidly and accurately evaluating the diversity of the ecological fishery resources on the large water surface according to claim 2, wherein the method comprises the following steps: typical waters include areas of heterogeneity, reservoir inlets, outlets, near shore and near center regions.
4. The method for rapidly and accurately evaluating the diversity of the ecological fishery resources on the large water surface according to claim 1, wherein the method comprises the following steps: the samples collected include water samples and sediment samples.
5. The method for rapidly and accurately evaluating the diversity of the ecological fishery resources on the large water surface according to claim 4, wherein the method comprises the following steps: collecting water in reservoir including upper layer water, taking water amount of 0.5-1L, and suction filtering with 0.45um mixed cellulose ester membrane filter membrane or suction filtering in laboratory within 24h after refrigeration.
6. The method for rapidly and accurately evaluating the diversity of the ecological fishery resources on the large water surface according to claim 4, wherein the method comprises the following steps: in the sediment sample collection, a mud sampler is used for collecting sediments in a water sampling place, and the sediments are respectively stored in an aseptic sealable wide-mouth bottle after passing through large granular substances in a solarization and rolling filter tank, wherein the large granular substances comprise large stones and plant branches.
7. The method for rapidly and accurately evaluating the diversity of the ecological fishery resources on the large water surface according to claim 1, wherein the method comprises the following steps: the sampling device needs to be cleaned before each use in sample collection to avoid cross infection.
8. The method for rapidly and accurately evaluating the diversity of the ecological fishery resources on the large water surface according to claim 1, wherein the method comprises the following steps: respectively adding lysis buffer solution and proteinase K into sample tissues in the environmental DNA extraction and amplification, and extracting the DNA of a water sample or sediment sample by using a DNA extraction kit after water bath lysis at 56 ℃; the NDA extraction kit comprises any one of a Power Water DNA extraction kit and a DNeasy Blood and Tissue DNA extraction kit; sample DNA is amplified by using primers designed from any one of the sequences of mitochondrial cytochrome b, 12S, 16S or COI of the species in the local water area.
9. The method for rapidly and accurately evaluating the diversity of the ecological fishery resources on the large water surface according to claim 1, wherein the method comprises the following steps: high throughput sequencing and macro barcode detection: and (3) freezing the amplified sequences by liquid nitrogen, quickly transporting the sequences to a biological company by adding dry ice for high-throughput sequencing and macro-barcode detection, and annotating the species information represented by each sequence.
10. The method for rapidly and accurately evaluating the diversity of the ecological fishery resources on the large water surface according to claim 1, wherein the method comprises the following steps: analyzing the diversity of fishery resources: and analyzing the diversity of fishery resources according to the compared species information by using the relative abundance of species, Alpha diversity, Shannon-Wiener index, Simpson index and Chaol index.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210824479.5A CN115044663A (en) | 2022-07-14 | 2022-07-14 | Method for rapidly and accurately evaluating diversity of ecological fishery resources on large water surface |
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| CN202210824479.5A CN115044663A (en) | 2022-07-14 | 2022-07-14 | Method for rapidly and accurately evaluating diversity of ecological fishery resources on large water surface |
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| CN202210824479.5A Pending CN115044663A (en) | 2022-07-14 | 2022-07-14 | Method for rapidly and accurately evaluating diversity of ecological fishery resources on large water surface |
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| CN (1) | CN115044663A (en) |
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- 2022-07-14 CN CN202210824479.5A patent/CN115044663A/en active Pending
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