Journal of Insect Biodiversity and Systematics

Journal of Insect Biodiversity and Systematics

Morphological and Molecular characterization of Mealybugs (Hemiptera: Coccomorpha: Pseudococcidae) Associated with Hibiscus rosa-sinensis (Malvaceae)

Document Type : Full-length Research Article

Authors
1 Department of Plant Protection, Faculty of Agriculture, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
2 Department of Entomology, Bangladesh Jute Research Institute, Manik Mia Avenue, Dhaka-1207, Bangladesh
3 Department of Crop science, Faculty of Agriculture, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
10.48311/jibs.12.03.633
Abstract
Complexities of the identification of mealybugs may compromise effective management practice for these pests. Hibiscus species (Malvaceae) are frequently attacked by different mealybug species globally. Nevertheless, little information is available concerning the mealybug fauna associated with H. rosa-sinensis in Malaysia. The aim of this study was to identify the mealybug species infesting the H. rosa-sinensis by morphological and DNA barcoding. The DNA barcoding study was performed by comparing the sequencing of the universal barcode DNA region, cytochrome c oxidase subunit I (COI), and ribosomal (18S and 28S). The nucleotide sequences confirmed the mealybug species, with the closest BLAST match in the GenBank queries sharing above 97% support. Based on taxonomic characterization by using scanning electron microscopy and DNA barcoding, three mealybug species, Paracoccus marginatus, Maconellicoccus hirsutus, and Phenacoccus solenopsis were confirmed in this study. DNA barcoding, along with the detailed external ultra-structural analysis of the mealybugs, could add a new dimension for identifying the mealybug species rapidly and more accurately. Ultimately, this study will help in decision-making in management practice for mealybugs.

Graphical Abstract

Morphological and Molecular characterization of Mealybugs (Hemiptera: Coccomorpha: Pseudococcidae) Associated with Hibiscus rosa-sinensis (Malvaceae)
Keywords
Subjects

Abbas, G., Arif, M.J., Saeed, S. & Karar, H. (2009) A new invasive species of genus Phenacoccus Cockerell attacking cotton in Pakistan. International Journal of Agriculture and Biology, 11 (1), 54–58.
Ahmed, M.Z., Ma, J.U.N., Qiu, B., He, R., Wu, M., Liang, F.A.N., Zhao, J., Lin, L.I., Hu, X., Lv, L., Breinholt, J.W. & Lu, Y. (2015) Genetic record for a recent invasion of Phenacoccus solenopsis (Hemiptera: Pseudococcidae) in Asia. Environmental Entomology, 44 (3), 907–918. https://doi.org/10.1093/ee/nvv034
Akintola, A.J. & Ande, A.T. (2008) First record of Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) on Hibiscus rosa-sinensis in Nigeria. Agricultural Journal, 3 (1), 1–3.
Ashfaq, M., Noor, A.R. & Mansoor, S. (2010) DNA-based characterization of an invasive mealybug (Hemiptera: Pseudococcidae) species damaging cotton in Pakistan. Applied Entomology and Zoology, 45 (3), 395–404.  https://doi.org/10.1303/aez.2010.395
Beltrà, A., Soto, A. & Malausa, T. (2012) Molecular and morphological characterisation of Pseudococcidae surveyed on crops and ornamental plants in Spain. Bulletin of Entomological Research, 102 (2), 165–172.  https://doi.org/10.1017/S0007485311000514
Bisevac, L. (1997) A new method for mounting thrips (Thysanoptera) on slides. Australian Journal of Entomology, 36 (3), 220. https://doi.org/10.1111/j.1440-6055.1997.tb01457.x
Boyer, S., Blakemore, R.J. & Wratten, S.D. (2011) An integrative taxonomic approach to the identification of three new New Zealand endemic earthworm species (Acanthodrilidae, Octochaetidae: Oligochaeta). Zootaxa, 2994 (1), 21–32. https://doi.org/10.11646/zootaxa.2994.1.2
Charles, J.G., Froud, K.J. & Henderson, R.C. (2000) Morphological variation and mating compatibility within the mealybugs Pseudococcus calceolariae and P. similans (Hemiptera: Pseudococcidae), and a new synonymy. Systematic Entomology, 25, 285–294. https://doi.org/10.1046/j.1365-3113.2000.00104.x
Chatzidimitriou, E., Simonato, M., Watson, G.W., Martinez-Sañudo, I., Tanaka, H., Zhao, J. & Pellizzari, G. (2016) Are Phenacoccus solani Ferris and P. defectus Ferris (Hemiptera: Pseudococcidae) distinct species. Zootaxa, 4093 (4), 539–551. https://doi.org/10.11646/zootaxa.4093.4.5
Correa, M., Aguirre, C., Germain, J.-F., Hinrichsen, P., Zaviezo, T., Malausa, T. & Prado, E. (2011) A new species of Pseudococcus (Hemiptera: Pseudococcidae) belonging to the "Pseudococcus maritimus" complex from Chile: molecular and morphological description. Zootaxa, 2926 (1), 46–54. https://doi.org/10.11646/zootaxa.2926.1.2
Cox, J.M. & Pearce, M.J. (1983) Wax produced by dermal pores in three species of mealybug (Homoptera: Pseudococcidae). International Journal of Insect Morphology and Embryology, 12 (4), 235–248. https://doi.org/10.1016/0020-7322(83)90020-X
Downie, D.A. & Gullan, P.J. (2004) Phylogenetic analysis of mealybugs (Hemiptera: Coccoidea: Pseudococcidae) based on DNA sequences from three nuclear genes, and a review of the higher classification. Systematic Entomology, 29 (2), 238–260. https://doi.org/10.1111/j.0307-6970.2004.00241.x
EFSA Panel on Plant Health (PLH), Bragard, C., Baptista, P., Chatzivassiliou, E., Di Serio, F., Gonthier, P., Jaques Miret, J.A., Justesen, A.F., Sven Magnusson, C., Milonas, P., Navas-Cortes, J.A., Parnell, S., Potting, R., Reignault, P.L., Stefani, E., Thulke, H.H., Van der Werf, W., Civera, A.V., Yuen, J., Zappalà, L., Gregoire, J.C., Malumphy, C., Antonatos, S., Kertesz, V., Maiorano, A., Papachristos, D. & MacLeod, A. (2022) Pest categorisation of Maconellicoccus hirsutusEFSA journal. European Food Safety Authority, 20 (1), e07024. https://doi.org/10.2903/j.efsa.2022.7024
 Felsenstein, J. (1985) Confidence Limits on Phylogenies: An Approach Using the Bootstrap. Evolution, 39 (4), 783. https://doi.org/10.2307/2408678
García Morales, M., 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, 2016, bav118. https://doi.org/10.1093/database/bav118
Hajibabaei, M., Janzen, D.H., Burns, J.M., Hallwachs, W. & Hebert, P.D.N. (2006) DNA barcodes distinguish species of tropical Lepidoptera. Proceedings of the National Academy of Sciences, 103 (4), 968–971.https://doi.org/10.1073/pnas.0510466103
Hall, T.A. (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41, 95–98.
Hardy, N.B., Gullan, P.J. & Hodgson, C.J. (2008) A subfamily level classification of mealybugs (Hemiptera: Pseudococcidae) based on integrated molecular and morphological data. Systematic Entomology, 33 (1), 51–71.  https://doi.org/10.1111/j.1365-3113.2007.00408.x
 Hazarika, M., Dutta, S.K., Bhattacharryya, B. & Dey, D. (2024) Scanning electron microscopy based morphology of papaya mealybug [Paracoccus marginatus Williams and Granara De Wilink (Hemiptera: Pseudococcidae)] on Mulberry plant of Assam, India. Journal of Advances in Biology & Biotechnology, 27 (10), 587–595. https://doi.org/10.9734/jabb/2024/v27i101481
Hebert, P.D.N., Cywinska, A., Ball, S.L. & DeWaard, J.R. (2003) Biological identifications through DNA barcodes. Proceedings of the Royal Society of London. Series B: Biological Sciences, 270 (1512), 313–321. https://doi.org/10.1098/rspb.2002.2218
Hodgson, C., Abbas, G., Arif, M.J., Saeed, S. & Karar, H. (2008) Phenacoccus solenopsis Tinsley (Sternorrhyncha: Coccoidea: Pseudococcidae), an invasive mealybug damaging cotton in Pakistan and India, with a discussion on seasonal morphological variation. Zootaxa, 191, 31–35. https://doi.org/10.11646/zootaxa.1913.1.1
Jhala, R.C., Bharpoda, T.M. & Patel, M.G. (2008) Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae), the mealybug species recorded first time on cotton and its alternate host plants in Gujarat, India. Uttar Pradesh Journal of Zoology, 28 (3), 403–406.
Kaila, L. & Ståhls, G. (2006) DNA barcodes: Evaluating the potential of COI to differentiate closely related species of Elachista (Lepidoptera: Gelechioidea: Elachistidae) from Australia. Zootaxa, 1170 (1), 1–26.  https://doi.org/10.11646/zootaxa.1170.1.1
Kondo, T., Gullan, P.J. & Williams, D.J. (2008) Coccidology. The study of scale insects (Hemiptera: Sternorrhyncha: Coccoidea). Ciencia y Tecnología Agropecuaria, 9 (2), 55–61. https://doi.org/10.21930/rcta.vol9_num2_art:118
Kumar, L.S., Sawant, A.S., Gupta, V.S. & Ranjekar, P.K. (2001) Genetic variation in Indian populations of Scirpophaga incertulas as revealed by RAPD-PCR analysis. Biochemical Genetics, 39, 43–57. https://doi.org/10.1023/A:1002797219182
Kumar, S., Stecher, G., Li, M., Knyaz, C. & Tamura, K. (2018) MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution, 35 (6), 1547–1549. https://doi.org/10.1093/molbev/msy096
Kumar, V., Tewari, S.K. & Datta, R.K. (1997) Dermal pores and wax secretion in mealybug Maconellicoccus hirsutus (Hemiptera, Pseudococcidae). A pest of mulberry. Italian Journal of Zoology, 64 (4), 307–311. https://doi.org/10.1080/11250009709356218
Langhoff, P., Authier, A., Buckley, T.R., Dugdale, J.S., Rodrigo, A. & Newcomb, R.D. (2009) DNA barcoding of the endemic New Zealand leafroller moth genera, Ctenopseustis and Planotortrix. Molecular Ecology Resources, 9 (3), 691–698.   https://doi.org/10.1111/j.1755-0998.2009.02537.x
Li, H.-M., Deng, R.-Q., Wang, J.-W., Chen, Z.-Y., Jia, F.-L. & Wang, X.-Z. (2005) A preliminary phylogeny of the Pentatomomorpha (Hemiptera: Heteroptera) based on nuclear 18S rDNA and mitochondrial DNA sequences. Molecular Phylogenetics and Evolution, 37 (2), 313–326. https://doi.org/10.1016/j.ympev.2005.07.013
Lin, C.-P. & Danforth, B.N. (2004) How do insect nuclear and mitochondrial gene substitution patterns differ? Insights from Bayesian analyses of combined datasets. Molecular Phylogenetics and Evolution, 30(3), 686-702. https://doi.org/10.1016/S1055-7903(03)00241-0
Linares, M.C., Soto-Calderón, I.D., Lees, D.C. & Anthony, N.M. (2009) High mitochondrial diversity in geographically widespread butterflies of Madagascar: a test of the DNA barcoding approach. Molecular Phylogenetics and Evolution, 50 (3), 485–495. https://doi.org/10.1016/j.ympev.2008.11.008
Liu, J., Li, Q., Kong, L., Yu, H. & Zheng, X. (2011) Identifying the true oysters (Bivalvia: Ostreidae) with mitochondrial phylogeny and distance based DNA barcoding. Molecular Ecology Resources, 11 (5), 820–830.  https://doi.org/10.1111/j.1755-0998.2011.03025.x
Magdalita, P.M. & Pimentel, R.B. (2010) Hibiscus Hybrids and Philippines' Women Achievers. National Academy of Science and Technology, Bicutan, Taguig City, Metro Manila. 58 pp.
Malausa, T., Fenis, A., Warot, S., Germain, J.-F., Ris, N., Prado, E., Botton, M., Vanlerberghe-Masutti, F., Sforza, R. & Cruaud, C. (2011) DNA markers to disentangle complexes of cryptic taxa in mealybugs (Hemiptera: Pseudococcidae). Journal of Applied Entomology, 135 (1–2), 142–155. https://doi.org/10.1111/j.1439-0418.2009.01495.x
Mastoi, M.I., Nur Azura, A., Muhammad, R., Idris, A.B. & Ibrahim, Y. (2011) First report of papaya mealybug Paracoccus marginatus (Hemiptera: Pseudococcidae) from Malaysia. Australian Journal of Basic and Applied Sciences, 5 (7), 1247–1250.
McDonnell, R.J., Rugman-Jones, P., Backeljau, T., Breugelmans, K., Jordaens, K., Stouthamer, R., Paine, T. & Gormally, M. (2011) Molecular identification of the exotic slug Arion subfuscus sensu stricto (Gastropoda: Pulmonata) in California, with comments on the source location of introduced populations. Biological Invasions, 13, 61–66. https://doi.org/10.1007/s10530-010-9789-5
 Miller, D.R. & Miller, G.L. (2002) Redescription of Paracoccus marginatus Williams and Granara de Willink (Hemiptera: Coccoidea: Pseudococcidae), including descriptions of the immature stages and adult male. Proceedings of the Entomological Society of Washington, 104 (1), 1–23.
Miller, D.R., Rung, A. & Parikh, G. (2014) Scale Insects, edition 2, a tool for the identification of potential pest scales at USA ports-of-entry (Hemiptera, Sternorrhyncha, Coccoidea). ZooKeys 431, 61–67. https://doi.org/10.3897/zookeys.431.7474
Mohammed, M.A., Aman-Zuki, A., Buang, M.G., Hamdan, N.H., Sani, M., King, P.J.H. & Yaakop, S. (2022) DNA barcode of mealybugs (Hemiptera: pseudococcidae) and ants species (Hymenoptera: formicidae) that associated with pineapple in Bintulu, Sarawak, Malaysia. Serangga, 27 (2), 38–53.
OEPP/EPPO. (2006) Maconellicoccus hirsutus. EPPO Bulletin, 36 (1), 167–169. https://doi.org/10.1111/j.1365-2338.2006.00921.x
Pacheco Da Silva, V.C., Bertin, A., Blin, A., Germain, J.F., Bernardi, D., Rignol, G., Botton, M. & Malausa, T. (2014) Molecular and morphological identification of mealybug species (Hemiptera: Pseudococcidae) in Brazilian vineyards. PLoS ONE, 9 (7), 1–13. https://doi.org/10.1371/journal.pone.0103267
Palma-Jimenez, M. & Blanco-Meneses, M. (2016) First study about morphological and molecular identification of Pseudococcus elisae (Hemiptera: Pseudococcidae) in Costa Rica. International Journal of Current Research, 8 (5), 31791–31800.
Ricupero, M., Biondi, A., Russo, A., Zappal, L. & Mazzeo, G. (2021) The cotton mealybug is spreading along the Mediterranean: First pest detection in Italian tomatoes. Insects, 12 (8), 675. https://doi.org/10.3390/insects12080675
Rugman-Jones, P.F., Morse, J.G. & Stouthamer, R. (2009) Rapid molecular identification of armored scale insects (Hemiptera: Diaspididae) on Mexican 'Hass' avocado. Journal of Economic Entomology, 102 (5), 1948–1953.  https://doi.org/10.1603/029.102.0527
Rung, A., Scheffer, S.J., Evans, G. & Miller, D. (2008) Molecular identification of two closely related species of mealybugs of the genus Planococcus (Homoptera: Pseudococcidae). Annals of the Entomological Society of America, 101 (3), 525–532. https://doi.org/10.1603/0013-8746(2008)101[525:MIOTCR]2.0.CO;2
Sartiami, D., Watson, G.W., Mohamad Roff, M. & Idris, A.B. (2016) Mealybugs (Hemiptera: Coccomorpha: Pseudococcidae) attacking Hibiscus rosa-sinensis L. in Malaysia, with two new country records. AIP Conference Proceedings. April 13–14, 2016. Faculty of Science and Technology, Universiti Kebangsaan Malaysia; 1784, 060007 pp. https://doi.org/10.1063/1.4966845
Sarvananda, L. (2018) Short introduction of DNA barcoding. International Journal of Research, 5 (4), 673–685.
Simon, C., Frati, F., Beckenbach, A., Crespi, B., Liu, H. & Flook, P. (1994) Evolution, weighting, and phylogenetic utility of mitochondrial gene sequences and a compilation of conserved polymerase chain reaction primers. Annals of the Entomological Society of America, 87 (6), 651–701. https://doi.org/10.1093/aesa/87.6.651
Singh, B.B. & Singh, R. (2014) Major rice insect pests in Northeastern UP. International Journal of Life Sciences, Biotechnology and Pharma Research, 1 (3), 124–143.
Sirisena, U., Watson, G.W., Hemachandra, K.S., Sage, O. & Wijayagunasekara, H.N.P. (2015) Scanning electron microscopy of six selected mealybug (Hemiptera: Pseudococcidae) species of Sri Lanka. Tropical Agricultural Research, 26 (2), 237–247. https://doi.org/10.4038/tar.v26i2.8088
Tamura, K. & Nei, M. (1993) Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Molecular Biology and Evolution, 10 (3), 512–526.
Williams, D.J. (1996) A brief account of the hibiscus mealybug Maconellicoccus hirsutus (Hemiptera: Pseudococcidae), a pest of agriculture and horticulture, with descriptions of two related species from southern Asia. Bulletin of Entomological Research, 86 (5), 617–628. https://doi.org/10.1017/S0007485300039420
Williams, D.J. (2004) Mealybugs of southern Asia. The Natural History Museum & Southdene, Kuala Lumpur. 896 pp.
Williams, D.J. & Granara, de W. (1992) Mealybugs of Central and South America. CAB International, Wallingford. 635 pp.
Wu, F., Liu, Z., Shen, H., Yu, F., Ma, J., Hu, X. & Zeng, L. (2014) Morphological and molecular identification of Paracoccus marginatus (Hemiptera: Pseudococcidae) in Yunnan, China. Florida Entomologist, 97 (4), 1469–1473.  https://doi.org/10.1653/024.097.0422
Zhang, Z.S., Qi, M., Guo, S. & Ma, X. (1988) Study on wax secretion capacity of the white wax insect (Ericerus pela Chavannes) in China. Zoological Research, 9 (01), 93–97.
 Zurovcova, M., Havelka, J.A.N., Starý, P., Vechtova, P., Chundelova, D., Jarosova, A. & Kucerova, L. (2010) " DNA barcoding" is of limited value for identifying adelgids (Hemiptera: Adelgidae) but supports traditional morphological taxonomy. European Journal of Entomology, 107 (2), 147. https://doi.org/10.14411/eje.2010.020

Articles in Press, Accepted Manuscript
Available Online from 16 June 2026

  • Receive Date 28 January 2026
  • Accept Date 10 June 2026
  • Publish Date 16 June 2026