Skip to main content
SpringerLink
Log in

Distinguishing Palm White Grub Complex, Leucopholis spp. (Coleoptera: Scarabaeidae: Melolonthinae) From India Using High-Resolution Melting (HRM) Analyses

  • Original Research Article
  • Published:
International Journal of Tropical Insect Science Aims and scope Submit manuscript

Abstract

This study aimed to identify the differences among three species of palm white grubs at morphological, ethological, ecological and molecular levels to develop a tool to aid in distinguishing the three species. A comprehensive survey was conducted in palm gardens in southern India. Third-instar larvae and adults of Leucopholis spp. were collected and brought into the laboratory for species-level identification using morphological traits. DNA was extracted and PCR-based amplification of cytochrome oxidase I (CO1) gene of the three Leucopholis spp. was undertaken, and the amplicons were sequenced. High-resolution melt (HRM) curve analysis was then attempted to differentiate the three Leucopholis spp. The survey indicated the predominance of Leucopholis lepidophora Blanchard in high ranges of Karnataka and Kerala States, Leucopholis burmeisteri Brenske in Dakshina Kannada district of Karnataka State and Leucopholis coneophora Burmeister along the West coast of Kerala State in India. The three species exhibited morphological differences in respect of chaetotaxy in the rastor of third instar larva and ethological differences with respect to flight behavior and congregation patterns. Multiple sequence alignment of the CO1 sequences revealed the presence of single nucleotide polymorphism (SNPs) between the three species. High-resolution melt (HRM) curve analysis was used to differentiate the three Leucopholis spp. utilizing primers designed based on single nucleotide polymorphisms (SNPs) identified in the CO1 sequences between the three species.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Data availability

All the sequences generated in the present study have been deposited in NCBI (Genbank accession numbers MG717658-MG717676). This has been indicated in the manuscript.

References

  • Abraham VA (1983) Biology, bionomics and control of coconut cock chafer, LeucopholisconeophoraBurm. Ph.D. Thesis Kerala Agricultural University Vellanikkara, p 215

  • Abraham VA (1993) Behaviour of adult beetles of the coconut white grub LeucopholisconeophoraBurm. Indian Coconut J 24:2–4

    Google Scholar 

  • Abraham VA, Kurian C (1970) Save your coconut palms from the ravages of white grubs. Coconut Bull 1:3–4

    Google Scholar 

  • Adsule VM, Patil SM (1994) Biology of sugar cane white grub L. lepidophora Bl. (Coleoptera: Scarabaeidae: Melolonthinae). Indian Sugar 44(1):25–28

    Google Scholar 

  • Ajamma YU, Mararo E, Omondi D, Onchuru T, Muigai AW, Masiga D and Villinger J (2016) Rapid and high throughput molecular identification of diverse mosquito species by high resolution melting analysis. F1000 Research 5

  • Avise JC (1994) Molecular markers. Natural history and evolution. Chapman and Hall, New York

  • Bellotti A and Schoonhoven A (1979) Miscellaneous pests of cassava. Cassava programme. CIAT, Columbia, pp 233- 236

  • Blanchard CE (1845) Histoire des Insectes, Traitant de leurs Moeurs et de leurs Metamorphoses en General et Comprenantune Nouvelle Classification Fonde e surleurs Rapports Naturels. Hymenopteres et Coleopteres, Part 1. Firmin Didot Freres, Paris, France.

  • Blanchard CE (1850) Ordre des Coleoptera. In: Edwards H, Blanchard CE, Lucus H (eds) Museum d Histoire Naturelle de Paris. Catalogue de la Collection Entomologique. Classe des Insectes, vol. 1, part 1. Milne Gide and Baudry, Paris, France

  • Bracalini M, Cerboneschi M, Croci F, Panzavolta T, Tiberi R, Biancalani C, Macconi S, Tegli S (2015) Alien pest molecular diagnostics: Can DNA traces be exploited to assess the damage caused by the western conifer seed bug on stone pine fructification. Bull Insectol 68(1):51–60

    Google Scholar 

  • Brenske E (1892) Beitragzurkennties der GattungenLepidiota and Leucopholis. Berliner EntomolZeitchri 37:33–61

    Article  Google Scholar 

  • Brenske E (1894) Die Melolothide der Palaearctischen and Orientalischen Region in Koniglichennaturhistorischen Museum zu Brussel. BeschreibungneuerArten und Bemerkungzubekannten. Mémoires De La Société Entomologique De Belgique 2:3–87

    Google Scholar 

  • Brown WM (1985) The mitochondrial genome of animals. In: MacIntyer RJ (ed) Molecular evolutionary Geneticsí, pp 95–130

  • Burmeister H (1855) Handbuch der Entomologie, volume 4, part 2. T.C.F. Enslin, Berlin, Germany

  • Chateigner-Boutin AL, Small I (2007) A rapid high-throughput method for the detection and quantification of RNA editing based on high-resolution melting of amplicons. Nucleic Acids Res 35(17):e114

    Article  PubMed  PubMed Central  Google Scholar 

  • Daniel M, Kumar TP (1976) Pests of arecanut. J Plant Crops 4:68–77

    Google Scholar 

  • Dejean C (1833) Catalogue des Coleopteres. DE LA Collection, Paris, p 160

    Google Scholar 

  • Dhami MK, Kumarasinghe L (2014) A HRM real-time PCR assay for rapid and specific identification of the emerging pest Spotted-Wing Drosophila (Drosophila suzukii). PLoS ONE 9(6):e98934

    Article  PubMed  PubMed Central  Google Scholar 

  • Kalleswaraswamy CM, Adarsha SK, Naveena NL, Sharanabasappa (2016) Adult emergence pattern and utilization of females as attractants for trapping males of white grubs, Leucopholisl epidophora (Coleoptera: Scarabaeidae), infesting arecanut in India. J Asia Pacific Entomol 15–22

  • Kapazoglou A, Andreadis SS, Drossou V, Madesis P, Savopoulou-Soultani M, Tsaftaris AS (2013) Cadherin Characterization and Cytochrome Oxidase (COI) HRM analysis in different geographical populations of the mediterranean corn borer, sesamia nonagrioides. J Agric Sci 6(1):173

    Google Scholar 

  • Klomtong P, Phasuk Y, Duangjinda M (2016) Animal species identification through high resolution melting real time PCR (HRM) of the Mitochondrial 16S rRNA Gene. Ann Anim Sci 16(2):415

    Article  CAS  Google Scholar 

  • Kumar ARV (1997) Biology and management of arecanut white grubs, Leucopholis spp. (Coleoptera : Scarabaeidae) in Karnataka. Ph.D. Thesis: Univ Agric Sci Bangalore, India p 230

  • Kumar TP (1974) Chemical control of arecanut white grubs Leucopholis burmeisteri Brenk. Arecanut Spices Bull 6:35

    Google Scholar 

  • Kumar TP, Daniel M (1981) Studies on the control of soil grubs of arecanut palm. Pesticides 15(9):29–30

    CAS  Google Scholar 

  • Lal SS and Pillai KS (1977) Occurrence of white grub, Leucopholis coneophora (Melolonthinae: Coleoptera) as a pest of tubers of Dioscoria alata and D. esculenta in Kerala. J Root Crops 3:69

  • Latreille PA (1802) Histoire naturelle des fourmis, etrecueil de memories etd’observationssur les abeilles, les araignees, les faucheurs, etautresinsects Par PA Latreille. del'imprimerie de Crapelet

  • Litsinger JA, Apostol RF, Obusan MB (1983) White grub Leucopholis irrorata (Coleoptera: Scarabaeidae): Pest status, Population dynamics, and chemical control in rice – maize cropping pattern in Philippines. J Econ Entomol 76:1133–1138

    Article  Google Scholar 

  • McLeay (1819) HoraeEntomologicae 1:209

  • Mahadevaswamy HM, Ramasamy A, Kalleshwaraswamy CM, Adarsha SK (2019) Arecanut white grubs Leucopholis species (Melolonthinae: Scarabaeidae: Coleoptera) morphological, molecular identification and phylogenetic analysis. J Asia Pacific Entomol 22:880–888

    Article  Google Scholar 

  • Malewski T, Draber-Mońko A, Pomorski J, Łoś M, Bogdanowicz W (2010) Identification of forensically important blowfly species (Diptera: Calliphoridae) by high-resolution melting PCR analysis. Int J Legal Med 124(4):277–285

    Article  PubMed  Google Scholar 

  • Nirula KK (1958) Investigations on the pests of coconut Part V. Leucopholisconeophora Burm. Indian Coconut J 12(1):10–34

    Google Scholar 

  • Nirula KK, Antony J, Menon KPV (1952) A new pest of coconut palm in India. Indian Coconut J 12(1):10–34

    Google Scholar 

  • Padmanabhan B, Daniel M (2003) Biology and bionomics of palm white grub Leucopholis burmeisteri. Indian J Entomol 65(4):444–452

    Google Scholar 

  • Pakendorf B, Stoneking M (2005) Mitochondrial DNA and human evolution. Annu Rev Genomics Hum Genet 6:165–183

    Article  CAS  PubMed  Google Scholar 

  • Pena VH, Fernández GJ, Gómez-Palacio AM, Mejía-Jaramillo AM, Cantillo O, Triana-Chávez O (2012) High-resolution melting (HRM) of the cytochrome B gene: a powerful approach to identify blood-meal sources in Chagas disease Vectors. PLOS Negl Trop Dis 6(2):e1530

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Prathibha PS (2015) Behavioural studies of palm white grubs Leucopholis spp. (Coleoptera: Scarabaeidae) and evaluation of new insecticides for their management. PhD thesis submitted to University of Agricultural sciences Bangalore, p 119

  • Prathibha PS Kumar ARV, Subaharan K (2013) Ethology of coconut root grub chafer Leucopholis coneophora Burmeister Melolonthinae : Scarabaeidae. Int J Agri Food Sci Technol

  • Prathibha PS, Subaharan K, Kumar ARV (2017) Toxicity and insecticide dissipation of applied in the management of arecanut white grub, Leucopholisburmeisteri Brenske. (Coleoptera: Scarabaeidae). Phytoparasitica 45:155–163

    Article  CAS  Google Scholar 

  • Prathibha PS, Subaharan K, Kumar ARV, Venugopal V (2018) Influence of abiotic factors on behaviour and adult emergence pattern of coconut white grub, Leucopholisconeophora Burmeister (Coleoptera: Scarabaeidae). Phytoparasitica 46:341–353

    Article  CAS  Google Scholar 

  • Puttarudraiah M, Channabasavanna GP (1957) Some new insect and mite pests of areca palm in Mysore. III. Arecanut 8(1):10–11

    Google Scholar 

  • Rajamani S, Nambiar KKN (1970) Pests of arecanut palm. Indian Fmg 20(8):23

    Google Scholar 

  • 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. Ann Entomol Soc Am 87:651–701

    Article  CAS  Google Scholar 

  • Swisher KD, Munyaneza JE, Crosslin JM (2012) High resolution melting analysis of the cytochrome oxidase I gene identifies three haplotypes of the potato psyllid in the United States. Environ Entomol 41(4):1019–1028

    Article  CAS  Google Scholar 

  • Thakur NSA, Firak DM, Behere GT, Firak PD, Saikia K (2012) Biodiversity of agriculturally important insects in North Eastern Himalaya an over view. Indian J Hill Farm 25(2):37–40

    Google Scholar 

  • Veeresh GK (1977) Studies on the root grubs in Karnataka, with special reference to bionomics and control of Holotrichia serrata F. (Coleoptera: Melolonthinae). UAS Monograph Ser. No.2, p 87

  • Veeresh GK (1981) Larval taxonomy of white grubs with special reference to Melolonthinae beetles. Final project report, ICAR Ad hoc Project on larval taxonomy of Melolonthinae beetles, 1977-80, Dept. of Entomology, UAS, Bangalore

  • Veeresh GK, Vijayaendra M, Reddy VN, Rajanna C, Rai PS (1982) Bio ecology and management of areca white grubs (Leucopholis spp.) (Melolonthinae : Scarabaeidae). J Soil Biol Eco 2:78–86

    Google Scholar 

  • Veeresh GK, Viswanath BN (1983) Crop losses due to white grubs. All India Seminar on Crop losses due to insect pests. Hyderabad, (Abstr) Section 18:5-6

  • Wetten A, Campbell C, Allainguillaume J (2016) High resolution melt and morphological analyses of mealybugs (Hemiptera: Pseudococcidae) from cacao: tools for the control of Cacao swollen shoot virus spread. Pest Manag Sci 72(3):527–533

    Article  CAS  PubMed  Google Scholar 

  • Wilhelm J, Pingoud A (2003) Real-time polymerase chain reaction. Chem Bio Chem 4:1120–1128

    Article  CAS  PubMed  Google Scholar 

  • Wilson AC, Cann RL, Carr SM, George M, Gyllensten UB, Helm-Bychowski KM, Higuchi RG, Palumbi SR, Prager EM, Sage RD, Stoneking M (1985) Mitochondrial DNA and tow perspectives on evolutionary genetics. Biol J Linn Soc 26:375–400

    Article  Google Scholar 

Download references

Acknowledgements

The authors thank Indian Council of Agricultural Research (ICAR) and Department of Biotechnology (Distributed Information sub-centre) for funding.

Author information

Authors and Affiliations

  1. Crop Protection Division, ICAR- Central Plantation Crops Research Institute, Kasaragod, Kerala, India, 671124

    P. S. Prathibha & V. S. Jilu

  2. Crop Improvement Division, ICAR- CPCRI, Kerala, 671124, Kasaragod, India

    M. K. Rajesh, A. A. Sabana & Vibina Venugopal

  3. Division of Germplasm Conservation and Utilization, ICAR- National Bureau of Agricultural Insect Resources, H.A. Farm Post, Bengaluru, Karnataka, India, 560024

    Kesavan Subaharan

Authors
  1. P. S. Prathibha
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. K. Rajesh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. A. Sabana
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kesavan Subaharan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Vibina Venugopal
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. V. S. Jilu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to P. S. Prathibha.

Ethics declarations

Competing interests

The authors have declared that they have no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Prathibha, P.S., Rajesh, M.K., Sabana, A.A. et al. Distinguishing Palm White Grub Complex, Leucopholis spp. (Coleoptera: Scarabaeidae: Melolonthinae) From India Using High-Resolution Melting (HRM) Analyses. Int J Trop Insect Sci 43, 1463–1474 (2023). https://doi.org/10.1007/s42690-023-01031-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s42690-023-01031-5

Keywords

Search

Navigation