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Molecular monitoring method for long-spiny starfish larvae in coral reef area

A technology for coral reefs and thorn sea stars, which is applied in the field of quantitative monitoring of biomolecular markers, and can solve problems such as monitoring of larvae of thorn sea stars that have not been seen.

Active Publication Date: 2021-09-03
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there is no method for monitoring the larvae of the long-spined starfish at home and abroad.

Method used

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  • Molecular monitoring method for long-spiny starfish larvae in coral reef area
  • Molecular monitoring method for long-spiny starfish larvae in coral reef area
  • Molecular monitoring method for long-spiny starfish larvae in coral reef area

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] (1) Preparation of standard reference DNA template of long spiny starfish larvae

[0023] Quantitatively take the larvae of the long spiny starfish and the brachybrachia larvae for suspension culture, collect about 10,000 samples of the brachycarpus larvae, and filter them with a 100 μm diameter pore / 50 mm diameter nylon mesh filter membrane (Millipore, Germany). Take out the filter membrane with sterile tweezers, and use The filter membrane was cut into pieces with sterile scissors, and together with the filter membrane, the DNA of the biological sample on the filter membrane was extracted using a bacterial genome DNA extraction kit (TaKaRa, China) to obtain the DNA of the larval sample. Take 1 μL of larval sample DNA (approximately 100 larval DNA copies / μL) for serial dilution (5-fold, 10-fold, 20-fold dilution) as a positive control template, and each μL positive control template represents 20, 10, and 5 larvae, respectively DNA copy.

[0024] (2) Filtration and col...

Embodiment 2

[0036] (1) Preparation of standard reference larval DNA template

[0037] Quantitatively take the larvae of the long spiny starfish and the brachybrachia larvae for suspension culture, collect about 10,000 samples of the brachycarpus larvae, and filter them with a 100 μm diameter pore / 50 mm diameter nylon mesh filter membrane (Millipore, Germany). Take out the filter membrane with sterile tweezers, and use The filter membrane was cut into pieces with sterile scissors, and together with the filter membrane, the DNA of the biological sample on the filter membrane was extracted using a bacterial genome DNA extraction kit (TaKaRa, China) to obtain the DNA of the larval sample. Take 1 μL of larval sample DNA (approximately 100 larval DNA copies / μL) for serial dilution (5-fold, 10-fold, 20-fold dilution) as a positive control template, and each μL positive control template represents 20, 10, and 5 larvae, respectively DNA copy.

[0038] (2) Filtration and collection of zooplankton ...

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Abstract

The invention discloses a molecular monitoring method for long-spiny starfish larvae in a coral reef area. The monitoring method comprises the following steps of 1) manufacturing a DNA template of standard reference long-spiny starfish larvae; 2) filtering and collecting zooplankton samples in the coral reef area; 3) extracting DNA of the zooplankton samples collected in the coral reef area; 4) carrying out PCR amplification and detection on the zooplankton samples. In the PCR detection, a pair of specific primers for the PCR detection of the long-spiny starfish are adopted. A forward primer is AcanP-TF:GCACCGATTTGTCTCTGCCAA. A reverse primer is AcanP-TR:AGTCCTTCTCTCCAGGT. If a target band exists at 785bp, the long-spiny starfish larvae is positive, and if the target band does not exist at the 785bp, the long-spiny starfish larvae is negative. According to the method, the long-spiny starfish larvae can be monitored, and the risk of outbreak of the population of the long-spiny starfish can be pre-warned in advance, so that the aim of early and timely prevention and control is fulfilled.

Description

Technical field: [0001] The invention belongs to the technical field of quantitative monitoring of biomolecular markers, and in particular relates to a molecular monitoring method for larvae of spiny starfish in coral reef areas. Background technique: [0002] Coral is an important biological resource in tropical oceans and is of great significance for maintaining marine ecosystems. The long-thorn starfish is an echinoderm that mainly feeds on corals. The long-thorn starfish has a large appetite and strong destructive power. Its population explosion can cause a large area of ​​coral reefs to die. Therefore, the long-thorn starfish is considered to be the most destructive enemy to corals. biology. How to prevent and control long-thorn starfish has become one of the most concerned issues for coral reef managers. In practice, it has been found that when a large number of long-thorn starfish adults appear, the coral reef has already been damaged from the bottom, so there is no...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2600/166C12Q2545/113Y02A40/81
Inventor 罗鹏李卓波夏淑蓉马波胡超群任春华
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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