欢迎访问《植物保护学报》网站！

首页 | 期刊简介 |
编委会
编委会
青年编委会
| 道德声明 | 投稿指南 | 联系我们 | 期刊订阅 | English

我国外来入侵生物福寿螺种类的多重PCR鉴别方法

引用本文：贺超,杨倩倩,刘苏汶,刘光富,俞晓平.我国外来入侵生物福寿螺种类的多重PCR鉴别方法.植物保护学报,2019,46(1):97-105

DOI：10.13802/j.cnki.zwbhxb.2019.2019912

摘要点击次数:

全文下载次数:

作者	单位	E-mail
贺超	中国计量大学生命科学学院, 浙江省生物计量及检验检疫重点实验室, 杭州 310018
杨倩倩	中国计量大学生命科学学院, 浙江省生物计量及检验检疫重点实验室, 杭州 310018	yqq@cjlu.edu.cn
刘苏汶	中国计量大学生命科学学院, 浙江省生物计量及检验检疫重点实验室, 杭州 310018
刘光富	中国计量大学生命科学学院, 浙江省生物计量及检验检疫重点实验室, 杭州 310018
俞晓平	中国计量大学生命科学学院, 浙江省生物计量及检验检疫重点实验室, 杭州 310018	yxp@cjlu.edu.cn

中文摘要:为研发能快速、准确地鉴别在我国为害严重的3种福寿螺——小管福寿螺Pomacea canaliculata、斑点福寿螺P.maculata和新发现入侵种Pomacea sp.的PCR技术，基于其线粒体全基因组序列，通过比对分析找到种间差异区段，分别设计并筛选了上述3种福寿螺的特异性引物对，建立了多重PCR检测方法，对其特异性和灵敏度进行评价。结果表明，所设计的引物特异性强，能从不同地理种群的小管福寿螺、斑点福寿螺和Pomacea sp.中分别扩增出1 238、901和571 bp的特异性条带，阴性对照无条带；退火温度为60℃时，32个扩增循环的检测灵敏度可达1 ng样品DNA量。所构建的多重PCR体系可应用于福寿螺不同部位组织碎片、不同性别及不同发育阶段样品的鉴别，具有准确快速、灵敏高效的特点，可用于植保检疫及食品安全部门福寿螺种类的检测。

中文关键词:福寿螺特异性引物分子鉴别线粒体基因多重PCR

Multiplex PCR detection of alien invasive apple snails introduced to China

Author Name	Affiliation	E-mail
He Chao	Zhejiang Provincial Key Laboratory of Biometrology and Inspection and Quarantine, College of Life Sciences, China Jiliang University, Hangzhou 310018, Zhejiang Province, China
Yang Qianqian	Zhejiang Provincial Key Laboratory of Biometrology and Inspection and Quarantine, College of Life Sciences, China Jiliang University, Hangzhou 310018, Zhejiang Province, China	yqq@cjlu.edu.cn
Liu Suwen	Zhejiang Provincial Key Laboratory of Biometrology and Inspection and Quarantine, College of Life Sciences, China Jiliang University, Hangzhou 310018, Zhejiang Province, China
Liu Guangfu	Zhejiang Provincial Key Laboratory of Biometrology and Inspection and Quarantine, College of Life Sciences, China Jiliang University, Hangzhou 310018, Zhejiang Province, China
Yu Xiaoping	Zhejiang Provincial Key Laboratory of Biometrology and Inspection and Quarantine, College of Life Sciences, China Jiliang University, Hangzhou 310018, Zhejiang Province, China	yxp@cjlu.edu.cn

Abstract:To develop fast and accurate PCR-based diagnostic methods for three apple snail species, i.e. Pomacea canaliculata, P. maculata and a newly discovered species Pomacea sp., which are damaging crops seriously in China, the species-specific primers for the three species based on the divergent sequences of their whole mitochondrial genomes were designed and screened, and a multiplex PCR was established and the specificity and sensitivity of the primers were assessed. The results showed that the primers were of high specificity, which were able to amplify 1 238, 901, and 571 bp fragments from P. canaliculata, P. maculata, and Pomacea sp., respectively. No amplicon was observed from the negative control. The detective limitation was 1 ng genomic DNA of any of P. canaliculata, P. maculata, and Pomacea sp. in the PCR assay at 60℃ annealing temperature for 32 cycles. This multiplex PCR with species-specific primers could be applied to detecting tissue fragments and samples of both sexes at various developmental stages for assisting species detection of apple snails in plant protection, plant quarantine and food security.

keywords:apple snail species-specific primer molecular diagnosis mitochondrial gene multiplex PCR

查看全文查看/发表评论下载PDF阅读器