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利用聚合酶链反应检测可可肿枝病毒及其在长蝽中的载体和假定载体并进行DNA条形码分析

Cacao swollen shoot virus detection and DNA barcoding of its vectors and putative vectors in L. by using polymerase chain reaction.

作者信息

Obok Ekemini Edet, Aikpokpodion Peter O, Ani Obinna Chukwu, Allainguillaume Joël, Wetten Andrew

机构信息

University of Calabar, Calabar, Nigeria.

Department of Biological, Biomedical and Applied Sciences, University of the West of England, Bristol, United Kingdom.

出版信息

BioTechnologia (Pozn). 2021 Sep 30;102(3):229-244. doi: 10.5114/bta.2021.108719. eCollection 2021.

Abstract

Cacao swollen shoot virus (CSSV) is an endemic pathogen causing significant economic losses to cacao ( L.) production in West Africa. There is limited updated report on the occurrence, spread, genetic diversity and species of CSSV and its mealybug vectors, especially in Nigeria. Nigeria is presently lagging behind in the search for resistance to CSSV and its vectors in L. The present study aimed to map and screen for the presence of CSSV and its natural vectors - female mealybugs (Pseudococcidae: Hemiptera) in cacao plantations in Nigeria. Symptomatic and asymptomatic cacao leaves and whole female mealybug samples were collected from major cacao-growing areas in Nigeria - Abia, Akwa Ibom, Cross River, Edo, Ondo and Oyo States. A total of 2568 cacao leaves from 1052 cacao trees were screened with polymerase chain reaction (PCR) using an open reading frame 1 (ORF 1) CSSV-specific primer pair. PCR screening of the mealybug species was performed using the cytochrome oxidase subunit I (COI) gene. A combination of scanning electron microscopy (SEM) and histology for morphological identification and DNA barcoding enabled to characterise the female mealybug species. The results revealed that CSSV and its mealybug vectors are present in the major cacaogrowing areas in Nigeria. Although CSSV and its vectors have been previously reported in Cross River, Ondo and Oyo States, our results present the first documented evidence of CSSV emergence and its mealybug vectors in Abia, Akwa Ibom and Edo States. We also present the first report of (Gimpel and Miller) mealybug species on cacao in Nigeria. In conclusion, it is pertinent to re-establish coordinated routine survey and monitoring of CSSV and its mealybug vector presence in L. in Nigeria.

摘要

可可肿枝病毒(CSSV)是一种地方性病原体,给西非的可可(Theobroma cacao L.)生产造成了重大经济损失。关于CSSV及其粉蚧传播媒介的发生、传播、遗传多样性和种类的最新报告有限,尤其是在尼日利亚。目前,尼日利亚在寻找可可对CSSV及其传播媒介的抗性方面落后。本研究旨在绘制并筛查尼日利亚可可种植园中CSSV及其天然传播媒介——雌性粉蚧(粉蚧科:半翅目)的存在情况。从尼日利亚主要的可可种植区——阿比亚州、阿夸伊博姆州、克罗斯河州、江户州、翁多州和奥约州采集了有症状和无症状的可可叶以及完整的雌性粉蚧样本。使用开放阅读框1(ORF 1)CSSV特异性引物对,通过聚合酶链反应(PCR)对来自1052棵可可树的总共2568片可可叶进行了筛查。使用细胞色素c氧化酶亚基I(COI)基因对粉蚧种类进行PCR筛查。结合扫描电子显微镜(SEM)和组织学进行形态鉴定以及DNA条形码技术,能够对雌性粉蚧种类进行特征描述。结果显示,CSSV及其粉蚧传播媒介存在于尼日利亚主要的可可种植区。尽管此前已在克罗斯河州、翁多州和奥约州报告过CSSV及其传播媒介,但我们的结果首次记录了阿比亚州、阿夸伊博姆州和江户州出现CSSV及其粉蚧传播媒介的证据。我们还首次报告了(金佩尔和米勒)粉蚧种类在尼日利亚可可上的情况。总之,重新建立对尼日利亚可可中CSSV及其粉蚧传播媒介存在情况的协调常规调查和监测很有必要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7026/9645568/f5f532cbe55d/BTA-102-3-45035-g001.jpg

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