A simple PCR-based method for detecting Anagyrus lopezi (Hymenoptera: Encyrtidae) and Prochiloneurus pulchellus (Hymenoptera: Encyrtidae), primary and hyper parasitoids of the cassava mealybug Phenacoccus manihoti (Hemiptera: Pseudococcidae)
{"title":"A simple PCR-based method for detecting Anagyrus lopezi (Hymenoptera: Encyrtidae) and Prochiloneurus pulchellus (Hymenoptera: Encyrtidae), primary and hyper parasitoids of the cassava mealybug Phenacoccus manihoti (Hemiptera: Pseudococcidae)","authors":"Shun-ichiro Takano, Ngoc Hung Nguyen, Thi Xuyen Le, Ah Nge Htwe, Keiji Takasu","doi":"10.1111/ens.12575","DOIUrl":null,"url":null,"abstract":"<p>Estimating parasitism rates in the field is essential for developing and evaluating biocontrol strategies using parasitoids. In this study, we developed a simple polymerase chain reaction (PCR)-based method for detecting parasitism of the cassava mealybug <i>Phenacoccus manihoti</i> Matile-Ferrero (Hemiptera: Pseudococcidae) by the primary parasitoid <i>Anagyrus lopezi</i> De Santis (Hymenoptera: Encyrtidae) and its hyperparasitoid <i>Prochiloneurus pulchellus</i> Silvestri (Hymenoptera: Encyrtidae). Primers were designed to amplify partial cytochrome <i>c</i> oxidase subunit I genes of each species, and their sensitivity was evaluated with mealybugs that had been parasitized by <i>A. lopezi</i> 0, 3, and 6 days earlier, and mummified mealybugs containing <i>A. lopezi</i> pupae that had been parasitized by <i>P. pulchellus</i> 0, 3, 6, 9, and 12 days earlier. The detection rate of parasitism by <i>A. lopezi</i> was 100% for all ages of <i>A. lopezi</i>. The detection rate of parasitism by <i>P. pulchellus</i> ranged from 94.1% to 100%, depending on its developmental stage. For <i>P. pulchellus</i>, template DNA was diluted 10 times before PCR because PCR with the original concentration showed low detection rates, presumably due to the presence of PCR inhibitors. Overall, our primers can be considered sufficiently sensitive to be used for detecting each species.</p>","PeriodicalId":11745,"journal":{"name":"Entomological Science","volume":null,"pages":null},"PeriodicalIF":0.7000,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Entomological Science","FirstCategoryId":"97","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/ens.12575","RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"ENTOMOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Estimating parasitism rates in the field is essential for developing and evaluating biocontrol strategies using parasitoids. In this study, we developed a simple polymerase chain reaction (PCR)-based method for detecting parasitism of the cassava mealybug Phenacoccus manihoti Matile-Ferrero (Hemiptera: Pseudococcidae) by the primary parasitoid Anagyrus lopezi De Santis (Hymenoptera: Encyrtidae) and its hyperparasitoid Prochiloneurus pulchellus Silvestri (Hymenoptera: Encyrtidae). Primers were designed to amplify partial cytochrome c oxidase subunit I genes of each species, and their sensitivity was evaluated with mealybugs that had been parasitized by A. lopezi 0, 3, and 6 days earlier, and mummified mealybugs containing A. lopezi pupae that had been parasitized by P. pulchellus 0, 3, 6, 9, and 12 days earlier. The detection rate of parasitism by A. lopezi was 100% for all ages of A. lopezi. The detection rate of parasitism by P. pulchellus ranged from 94.1% to 100%, depending on its developmental stage. For P. pulchellus, template DNA was diluted 10 times before PCR because PCR with the original concentration showed low detection rates, presumably due to the presence of PCR inhibitors. Overall, our primers can be considered sufficiently sensitive to be used for detecting each species.
期刊介绍:
Entomological Science is the official English language journal of the Entomological Society of Japan. The Journal publishes original research papers and reviews from any entomological discipline or from directly allied field in ecology, behavioral biology, physiology, biochemistry, development, genetics, systematics, morphology, evolution and general entomology. Papers of applied entomology will be considered for publication if they significantly advance in the field of entomological science in the opinion of the Editors and Editorial Board.