粉蚧科无损形态基因组DNA提取技术  

Extracting of Genomic DNA from Mealybugs(Hemiptera: Pseudococcidae) Without Conferring External Morphological Damage

作　　者：叶琳雄 符清流 胡荣[1] 李加慧[1] YE Linxiong FU Qingliu HU Rong LI Jiahui(College of Environment and Plant Protection, Hainan University, Haikou, Hainan 570228, Chin)

机构地区：[1]海南大学环境与植物保护学院,海口570228

出　　处：《热带生物学报》2016年第4期497-499,共3页Journal of Tropical Biology

基　　金：公益性行业(农业)科研专项(201403075);海南省自然科学基金项目(20163044);国家自然科学基金项目(31660129);国家质量监督检验检疫总局科技计划项目(2016IK094)

摘　　要：以双条拂粉蚧(Ferrisia virgata Cockerell)为研究对象,釆用蛋白酶K代替粉蚧玻片标本制作中的10%KOH用于组织消化,消化液用试剂盒法进行基因组DNA提取,剩余虫体进行玻片制作;同时比较了虫体全研磨、腹部微针扎孔与虫体不做任何处理共3种处理方法提取基因组DNA的效果。结果表明:3种处理方法均可成功提取基因组DNA,后2种方法处理后的剩余虫体可制作成玻片标本,实现了粉蚧无损形态的基因组DNA提取技术,为粉蚧的DNA条形码和系统发育研究提供了技术保障。Mealybug Ferrisia virgata Cockerell was digested with Proteinase K solution rather than 10% KOH to extract its genomic DNA. F. virgata was pretreated before extraction by grinding the whole mealybug,pricking the mealybug with micro-pins on abdomen or without any treatment to the mealybug. Genomic DNA was extracted by the digestion kit following its directions,and the rest of the mealybug left after digestion was used for slide preparation. The results show genomic DNA was extracted from the mealybug with all the three pre-treatments and that the rest of the mealybug left in the latter two pretreatments was good to make slides. This is a new protocol for genomic DNA extraction combined with permanent slides mounting in mealybugs. It would be useful for DNA barcoding and systematic research of mealybugs.

关 键 词：粉蚧 基因组DNA 提取 无损形态 DNA条形码 

分 类 号：S433.3[农业科学—农业昆虫与害虫防治]