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扶桑绵粉蚧Phenacoccus solenopsis Tinsley是我国近年新发现的一种严重威胁农林业生产的重要外来入侵害虫。针对扶桑绵粉蚧与其他粉蚧类昆虫难以准确快速识别且适生区广泛的问题,以扶桑绵粉蚧为靶标,以我国常见的其他7种粉蚧为参照,采用基于线粒体DNA细胞色素C氧化酶亚基Ⅰ(mtDNA COI)基因序列的种特异性(species-specific COI,SS-COI)PCR方法,研究其快速分子检测技术。通过已知粉蚧的COI基因序列设计通用型引物1对,获得扶桑绵粉蚧及其他7种粉蚧包括康氏粉蚧Pseudococcus comstocki Kuwana、南洋臀纹粉蚧Planococcus lilacius Cockerell、木槿曼粉蚧Maconellicoccus hirsutus(Green)、甘蔗红粉蚧Saccharicoccus sacchari(Cockerell)、新菠萝灰粉蚧Dysmicoccus neobrevipes Beardsley、番石榴粉蚧Planococcus minor Maskel和石蒜绵粉蚧Phenacoccus solani Ferris的COI基因序列,根据测序结果及数据库中已知粉蚧的COI基因序列设计SS-COI引物1对(PSZTF1/PSZTR1),其扩增片段大小为546bp。种特异性检验结果表明,该引物只对扶桑绵粉蚧的COI基因具有扩增能力,对其他7种粉蚧不具有扩增效果;该引物不仅对成虫具有良好的扩增能力,对不同虫态的扶桑绵粉蚧以及来自我国不同省市的14个地理种群和口岸截获的来自巴基斯坦的扶桑绵粉蚧亦具有同样的扩增效能。这些结果表明,该技术体系完全可用于扶桑绵粉蚧的准确识别及其检测监测,对有效阻截其进一步扩张蔓延意义重大。
Phenacoccus solenopsis Tinsley is an important invasive pest newly discovered in China in recent years that seriously threatens the production of agriculture and forestry. Fusong melanoid and other mealybugs insects are difficult to accurately and quickly identify and adapt to a wide range of problems in habitats, with Fupi Melatonin as a target, with our other common 7 mealybugs as a reference, based on mitochondrial DNA Cytochrome C (SS-COI) PCR method of mtDNA COI gene sequences to study its rapid molecular detection technology. A pair of universal primers was designed based on the COI gene sequence of known mealybugs. Seven mealybugs including Ficus comstocki Kuwana, Planococcus lilacius Cockerell, Maconellicoccus The COI gene sequences of hirsutus (Green), Saccharicoccus sacchari (Cockerell), Dysmicoccus neobrevipes Beardsley, Planococcus minor Maskel and Phenacoccus solani Ferris were determined according to the sequencing results and database Known mealybugs COI gene sequence design SS-COI primer 1 (PSZTF1 / PSZTR1), the amplified fragment size of 546bp. Species-specific test results showed that the primer only had the ability to amplify the COI gene of mealybug mealybugs and did not have any amplification effect on the other seven mealybugs. The primer not only had good ability to amplify the adult mealybugs, State Fuso mealybugs and 14 geographic populations and ports from different provinces and cities in China from Pakistan captured Fusong mealybugs also have the same amplification efficiency. These results show that the technology system can be completely used for the accurate identification and monitoring of mealybug mulberry mealybugs, which is of great significance to effectively intercepting and spreading its further expansion.