Rapid diagnosis of the invasive wax scale, Ceroplastes rusci Linnaeus (Hemiptera: Coccoidea: Coccidae) using nested PCR
Rapid diagnosis of the invasive wax scale, Ceroplastes rusci Linnaeus (Hemiptera: Coccoidea: Coccidae) using nested PCR
复制标题
使用巢式 PCR 快速诊断侵入性蜡鳞 Ceroplastes rusci Linnaeus（半翅目：球虫总科：球虫科）
DOI：
10.1111/jen.12155
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发表时间：
2015
影响因子：
1.9
通讯作者：
Wu S. -A.
中科院分区：
文献类型：
--
作者：
Deng J.;Wang X. -B.;Yu F.;Zhou Q. -S.;Bernardo U.;Zhang Y. -Z.;Wu S. -A.
无花果蜡蚧Ceroplastes rusci（Linnaeus）（半翅目：蚧总科：蚧科）是一种原产于非洲热带的入侵性果树害虫，对我国的果树生产具有潜在的威胁。C.由于缺乏易于区分的形态特征，rusci很难识别。建立了一种快速、准确、可靠的鉴定C.需要在检疫工作中使用rusci来检测进一步的传播。在本研究中，我们描述了一种巢式PCR方法的分子鉴定C. rusci。根据C. rusci和其他五种蜡蚧。从96 C成功扩增出200 bp的片段。rusci在中国和越南的7个地理种群中有13个个体，在意大利的2个种群中有13个个体。rusci）。这些特征条带在中国广泛分布的其它五种蜡蚧中均未观察到，即C. ceriferus（Fabricius）、蜡蝉C. florescence sis Comstock，C.格林绿色，C. pseudoceriferus绿色和C.鲁本斯·马斯凯尔。灵敏度测试表明，即使在约1.5 × 10−5 ng/μl的DNA模板浓度下，以及在成年雌性、单个一龄幼虫和卵的平均DNA模板浓度分别为14.7、6.3和3.0 ng/μl的情况下，也能产生诊断条带。我们的研究表明，C。rusci使用巢式PCR是快速和准确的，并显示在植物检疫程序的潜力。
The fig wax scale, Ceroplastes rusci (Linnaeus) (Hemiptera: Coccoidea: Coccidae), is an invasive fruit pest of Afrotropical origin and potentially could become a serious threat to commercial fruit crops in China. C. rusci is difficult to identify owing to the shortage of easily distinguishable morphological characters. A rapid, accurate and reliable method to identify C. rusci in quarantine work is needed to detect further spread. In the present study, we describe a nested PCR method for the molecular identification of C.rusci. The nested PCR primers were designed based on variations in the barcode region of COI sequences between C. rusci and five other Ceroplastes species. A 200‐bp fragment was successfully amplified from 96 C. rusci individuals of seven geographical populations in China and Vietnam, and 13 individuals of two populations in Italy (the type country for C. rusci). These provided diagnostic bands that were not observed in any of five other Ceroplastes species widely distributed in China, namely, C. ceriferus (Fabricius), C. floridensis Comstock, C. japonicus Green, C. pseudoceriferus Green and C. rubens Maskell. Sensitivity tests revealed that diagnostic bands were generated even with a DNA template concentration of ~1.5 × 10−5 ng/μl, and with average DNA template concentrations for adult females, single first‐instar nymphs and eggs of 14.7, 6.3 and 3.0 ng/μl, respectively. Our study demonstrates that the molecular diagnosis of C. rusci using nested PCR is rapid and accurate and shows potential in plant quarantine programmes.