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Journal of Integrative Agriculture  2019, Vol. 18 Issue (5): 1042-1049    DOI: 10.1016/S2095-3119(18)62043-X
Special Issue: 昆虫和植物互作合辑Insect and Plant Interact
Plant Protection Advanced Online Publication | Current Issue | Archive | Adv Search |
Species-specific COI primers for rapid identification of a globally significant invasive pest, the cassava mealybug Phenacoccus manihoti Matile-Ferrero
WANG Yu-sheng1, TIAN Hu2, WAN Fang-hao1, ZHANG Gui-fen1 
1 State Key Laboratory for Biology of Plant Diseases and Insect Pests/Key Laboratory of Integrated Pest Management of Crop, Ministry of Agriculture/Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China
2 Caofeidian Sub-Center of Hebei Entry-Exit Inspection and Quarantine Technical Center, Tangshan 063200, P.R.China
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The globally invasive cassava mealybug Phenacoccus manihoti Matile-Ferrero is a pernicious pest of cassava, and its recent introduction into Asia has raised considerable alarm.  To slow or prevent further invasion, an accurate, simple, and developmental-stage-independent detection method for P. manihoti is required.  In the present study, a PCR method based on a species-specific mitochondrial DNA cytochrome oxidase I (SS-COI) marker was developed for rapid identification of P. manihoti.  One pair of SS-COI primers (PMSSZW-1F and PMSSZW-1R) was designed based on sequence variations in the COI gene among P. manihoti and related mealybug species.  Specificity of the primer pair was validated on 21 closely related species.  Sensitivity tests were performed on four immature developmental stages and female adults.  Efficacy tests demonstrated that at the relatively low concentration of (135.2±14.7) pg μL–1 resuspended DNA, the specific fragment was detected in all replicates.  Furthermore, the SS-COI primer pair was assayed on three populations of P. manihoti from major exporting countries of cassava.  The PCR assay was proved to be a rapid, simple, and reliable molecular measure for the identification of P. manihoti.  This tool will be useful for quarantine, monitoring, and management of this invasive pest.
Keywords:  Phenacoccus manihoti        cassava mealybug        invasive pest        molecular identification        species-specific COI primers  
Received: 06 May 2018   Accepted:
Fund: This work was supported by the National Key R&D Program of China (2017YFC1200600, 2016YFC1201200 and 2015BAD08A16) and the Science and Technology Innovation Program of CAAS (caascx-2013-2018-IAS).
Corresponding Authors:  Correspondence ZHANG Gui-fen, Tel: +86-10-82109572, Fax: +86-10-82105927, E-mail:   
About author:  WANG Yu-sheng, E-mail:;

Cite this article: 

WANG Yu-sheng, TIAN Hu, WAN Fang-hao, ZHANG Gui-fen. 2019. Species-specific COI primers for rapid identification of a globally significant invasive pest, the cassava mealybug Phenacoccus manihoti Matile-Ferrero. Journal of Integrative Agriculture, 18(5): 1042-1049.

Bellotti A, Campo B V H, Hyman G. 2012. Cassava production and pest management: Present and potential threats in a changing environment. Tropical Plant Biology, 5, 39–72.
Bellotti A C, Smith L, Lapointe S L. 1999. Recent advances in cassava pest management. Annual Review of Entomology, 44, 343–370.
Black I V W C. 1993. PCR with arbitrary primers: Approach with care. Insect Molecular Biology, 2, 1–6.
Calatayud P A, Le Rü B P. 2006. Cassava-Mealybug Interactions. IRD éditions, Paris.
Cox M J, Williams D J. 1981. An account of cassava mealybugs (Hemiptera: Pseudococcidae) with a description of a new species. Bulletin of Entomological Research, 71, 247–458.
Herren H R. 1981. IITA’s role and action in controlling the cassava mealybug in Africa. IITA Research Briefs, 2, 1–4.
Herren H R, Neuenschwander P. 1991. Biological control of cassava pests in Africa. Annual Review of Entomology, 36, 257–283.
Hosseini R, Hajizadeh J. 2011. Molecular identification of three of the most important mealybug species (Hemiptera: Sternorrhyncha: Coccoidea: Pseudococcidae) on ornamental plants in Guilan Province, Iran. Zootaxa, 3009, 46–54.
Hosseini R, Keller M A, Schmidt O, Framenau V W. 2007. Molecular identification of wolf spiders (Araneae: Lycosidae) by multiplex polymerase chain reaction. Biological Control, 40, 128–135.
Jiang F, Li Z H, Deng Y L, Wu J J, Liu R S, Buahom N. 2013. Rapid diagnosis of the economically important fruit fly, Bactrocera correcta (Diptera: Tephritidae) based on a species-specific barcoding cytochrome oxidase I marker. Bulletin of Entomological Research, 103, 363–371.
Liebhold A M, Tobin P C. 2008. Population ecology of insect invasions and their management. Annual Review of Entomology, 53, 387–408.
Löhr B, Varela A M, Santos B. 1990. Exploration for natural enemies of the cassava mealybug, Phenacoccus manihoti (Homoptera: Pseudococcidae), in South America for the biological control of this introduced pest in Africa. Bulletin of Entomological Research, 80, 417–425.
Malumphy C. 2005. Insects and mites intercepted on fresh cassava (Manihot esculenta) foliage imported into England and Wales. Entomologist’s Monthly Magazine, 56, 53–63.
Matile-Ferrero D. 1978. Cassava mealybug in the People’s Republic of Congo. In: Nwanze K F, Leuschner K, eds., Proceedings of the International Workshop on the Cassava Mealybug Phenacoccus manihoti Mat.–Ferr. (Pseudococcidae). International Institute of Tropical Agriculture, Ibadan. pp. 29–46.
Morales M G, Denno B D, Miller D R, Miller G L, Ben-Dov Y, Hardy N B. 2016. ScaleNet: A literature-based model of scale insect biology and systematics. Database, doi: 10.1093/database/bav118
Muniappan R, Shepard B M, Watson G W. 2009. New records of invasive insects (Hemiptera: Sternorrhyncha) in Southeast Asia and West Africa. Journal of Agricultural and Urban Entomology, 26, 167–174.
Neuenschwander P, Herren H R, Harpaz I, Badulescu D, Akingbohungbe A E. 1988. Biological control of the cassava mealybug, Phenacoccus manihoti, by the exotic parasitoid Epidinocarsis lopezi in Africa. Philosophical Transactions of the Royal Society of London (Series B: Biological Sciences), 318, 319–333.
Nwanze K F. 1982. Relationships between cassava root yields and crop infestations by the mealybug, Phenacoccus manihoti. Tropical Pest Management, 28, 27–32.
Nweke F I. 2009. Controlling cassava mosaic virus and cassava mealybug in sub-Saharan Africa, In: Spielman D, Pandya-Lorch R, eds., Proven Successes in Agricultural Development. International Food Policy Research Institute, Washington. pp. 99–122.
Park D S, Jin L Y, Hahn K W, Suh S J, Hong K J, Oh H W. 2010. Molecular identification of mealybugs (Hemiptera: Pseudococcidae) found on Korean pears. Journal of Economic Entomology, 103, 25–33.
Park D S, Suh S J, Hebert P D N, Oh H W, Hong K J. 2011. DNA barcodes for two scale insect families, mealybugs (Hemiptera: Pseudococcidae) and armored scales (Hemiptera: Diaspididae). Bulletin of Entomological Research, 101, 429–434.
Parsa S, Kondo T, Winotai A. 2012. The cassava mealybug (Phenacoccus manihoti) in Asia: First records, potential distribution, and an identification key. PLoS ONE, 7, e47675.
Le Ru B, Tertuliano B. 1993. Tolerance of different host plants to the cassava mealybug Phenacoccus manihoti Matile-Ferrore (Homoptera: Pseudococcidae). International Journal of Pest Management, 39, 379–389.
Rugman-Jones P F, Hoddle M S, Mound L A, Stouthamer R. 2006. Molecular identification key for pest species of Scirtothrips (Thysanoptera: Thripidae). Journal of Economic Entomology, 99, 1813–1819.
Saccaggi D L, Krüger K, Pietersen G. 2008. A multiplex PCR assay for the simultaneous identification of three mealybug species (Hemiptera: Pseudococcidae). Bulletin of Entomological Research, 98, 27–33.
Shi J, Xie Y P, Xue J L, Yao G Q. 2005. Comparative study of methods for isolation of genomic DNA of scale insects. Chinese Bulletin of Entomology, 42, 207–211. (in Chinese)
Tang F D. 1992. The Pseudococcidae of China. China Agricultural Science and Technology Press, Beijing. (in Chinese)
Tian H, Li X F, Wan F H, Zhang G F, Zhang J L. 2013. Identification of Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) with species-specific COI (SS-COI) primers. Acta Entomologica Sinica, 56, 689–696. (in Chinese)
Wang Y S. 2016. Identification of common mealybug species based on double genes barcoding techniques in China. MSc thesis, Chinese Academy of Agricultural Sciences, Beijing. (in Chinese)
Winotai A, Goergen G, Tamò M, Neuenschwander P. 2010. Cassava mealybug has reached Asia. Biocontrol News and Information, 31, 10–11.
Wu S A, Wang Y P. 2011. Precaution of cassava mealybug, Phenacoccus manihoti Matile-Ferrero in China. Journal of Environmental Entomology, 33, 122–125. (in Chinese)
Yonow T, Kriticos D J, Ota N. 2017. The potential distribution of cassava mealybug (Phenacoccus manihoti), a threat to food security for the poor. PLoS ONE, 12, e0173265.
Zhang G F, Meng X Q, Min L, Qiao W N, Wan F H. 2012. Rapid diagnosis of the invasive species, Frankliniella occidentalis (Pergande): A species-specific COI marker. Journal of Applied Entomology, 136, 410–420.
Zhou X, Xu W, Wang C, He Y Y, Chen K, Zhang J Q, Yan J. 2014a. Potential geographic distribution of Phenacoccus manihoti Matile-Ferrero in China. Journal of Southern Agriculture, 45, 590–595. (in Chinese)
Zhou X, Zhang Q E, Pan X B, He Y Y, Huang Y, Yan J. 2014b. Study on the risk analysis of Phenacoccus manihoti Matile-Ferrero, on imported host products. Journal of Environmental Entomology, 36, 298–304. (in Chinese)
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