Specific primer pair for rapid identification of alien invasive Pomacea in China based on multiple PCR (Polymerase Chain Reaction) and application of specific primer pair

A specific primer pair and apple snail technology, applied in the biological field, can solve the problems of variation, uncertainty, and difficulty in identification of snail species, and achieve the effects of short time-consuming, high detection sensitivity, and easy operation

Pending Publication Date: 2016-05-25
CHINA JILIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

small tube apple snail P. canaliculata with spotted apple snails P. maculata As closely related species, they are highly similar in many aspects such as morphological characteristics and living habits; in addition, due to the high environmental adaptability of apple snails,

Method used

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  • Specific primer pair for rapid identification of alien invasive Pomacea in China based on multiple PCR (Polymerase Chain Reaction) and application of specific primer pair
  • Specific primer pair for rapid identification of alien invasive Pomacea in China based on multiple PCR (Polymerase Chain Reaction) and application of specific primer pair

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: the design of multiplex PCR specific primer pair

[0026] The present invention designs specific primer pairs for the two invasive apple snails, as shown in Table 1, on the basis of obtaining the DNA barcode segments of the mitochondrial COI gene DNA segments of two kinds of invasive apple snails.

[0027] Based on the common apple snail mitochondrial cytochrome oxidase subunit I barcode (mtDNACOIbarcode) segment, a multiplex PCR-specific primer pair that can identify two invasive apple snails was designed and synthesized:

[0028] PomF (sequence 1 of the sequence listing): 5'-GTTTACTTATTCGTGCTGAGTTAGG-3'; Tm=55.6°C, GC%=40%;

[0029] PcanR (sequence 2 of the sequence listing): 5'-GCAAGAACCGGCAATGATAAGAGCA-3'; Tm=64.1°C, GC%=48%.

[0030] PmacR (sequence 3 of the sequence listing): 5'-AGCTAAAGGAGGGTACACTGTTCAC-3'; Tm=60.3°C, GC%=48%.

Embodiment 2

[0031] Example 2: Application of specific primers to detect two invasive apple snails

[0032] Adopt following sample in the embodiment:

[0033] Sample 1-2: Apple snails collected from population 1 of Putuo Island, Zhoushan, Zhejiang P. canaliculata Egg grains of different egg masses; samples 3-4: apple snails collected from population 2 of Putuo Island, Zhoushan, Zhejiang P. canaliculata Eggs of different egg masses; samples 5-6: apple snails collected from Jieyang Airport District, Guangdong P. canaliculata Eggs of different egg masses; samples 7-8: apple snails collected from South China Agricultural University in Guangzhou, Guangdong P. canaliculata Egg grains of different egg masses; samples 9-10: apple snails collected from Yuyao, Zhejiang P. maculata Egg grains of different egg masses; samples 11-12: apple snails collected from Guangxi University in Nanning, Guangxi P. maculata Egg grains of different egg masses; samples 13-14: apple snails collected from Fuj...

example 3

[0053] Example 3: Detection of two invasive apple snail samples at different developmental stages

[0054] The samples used for the detection of two kinds of apple snail samples at different developmental stages included:

[0055] Sample 17: Apple snails collected from Tangpu Village, Jieyang Airport District, Guangdong P. canaliculata egg;

[0056] Sample 18: Apple snails collected from Tangpu Village, Jieyang Airport District, Guangdong P. canaliculata On the third day of hatching, young snails;

[0057] Sample 19: Apple snails collected from Tangpu Village, Jieyang Airport District, Guangdong P. canaliculata into a snail;

[0058] Sample 20: Apple snails collected from Yuyao, Zhejiang P. maculata egg;

[0059] Sample 21: Apple snails collected from Yuyao, Zhejiang P. maculata On the fifth day of hatching, young snails;

[0060] Sample 22: Apple snails collected from Yuyao, Zhejiang P. maculata Become a snail.

[0061] 1. Genomic DNA extraction

[0062] Take...

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Abstract

The invention discloses a multiple specific primer pair for assisting in identification and detection of alien invasive species of Pomacea canaliculata and Pomacea maculata and further discloses application of the multiple specific primer pair, aiming to overcome the difficulty in identifying and distinguishing Pomacea canaliculata and Pomacea maculata according to morphological characteristics because Pomacea canaliculata and Pomacea maculata are relative species with similar morphology. The specific primer pair is composed of DNA sequences expressed by a sequence 1, a sequence 2 and a sequence 3 of a sequence table. The specific primer pair can be applied to help to identity the alien invasive species of Pomacea canaliculata and Pomacea maculata molecularly without being affected by individual growth states or sample integrity, and has the advantages of high economic efficiency, convenience, accuracy and high sensitivity. The specific primer pair and the application thereof can be applied to plant protection departments and quarantine department in a wide scope and solves the problem about identification of the alien invasive species of Pomacea canaliculata and Pomacea maculata in China, which troubles the plant protection and quarantine departments for a long time.

Description

technical field [0001] The invention relates to a group of specific primer pairs for assisting in the identification of alien invasion organisms apple snails and applications thereof, belonging to the field of biotechnology. Background technique [0002] The apple snail belongs to the mollusca (Mollusca), gastropod (Gastropoda), new gastropod (Caeogastropoda), bottle snail family (Ampullariidae), apple snail ( Pomacea Perry, 1810). The apple snail was introduced and colonized in my country as a food in the early 1980s, but it was abandoned by people due to its poor taste, and it was bred in large numbers in the field; at present, the snail has rapidly expanded to Zhejiang, Fujian, Guangdong, Guangxi, and Hainan , Yunnan, Guizhou, Jiangxi, Sichuan, Chongqing, Hunan and Anhui and other provinces and caused disasters. Apple snails can harm a variety of aquatic crops and cause serious losses to rice production in southern my country; at the same time, apple snails can destroy t...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/16C12Q2537/143Y02A90/40
Inventor 杨倩倩俞晓平刘苏汶张蓬军许益鹏刘光富蒋家成
Owner CHINA JILIANG UNIV
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