INTEGRATIVE MORPHO-MOLECULAR ANALYSIS OF Papilio polytes, Papilio polymnestor, AND Euploea core FROM JHARKHAND (INDIA) USING ADVANCED BIOTECHNOLOGICAL AND BIOINFORMATIC APPROACHES Kumar Manoj. Ranjan Rakesh, Pratik Raj, Tirkey Suraj, Kumar Aman

Details: Hits: 129

ISSN 2410-7751 (Print)
ISSN 2410-776X (Online)

cover biotech acta general

Biotechnologia Acta Т. 18, No. 4, 2025
P. 46-59, Bibliography 44 , Engl.
UDC: 595.783:57.086:004.94(540.43)
doi: https://doi.org/10.15407/biotech18.04.046

Full text: (PDF, in English)

INTEGRATIVE MORPHO-MOLECULAR ANALYSIS OF Papilio polytes, Papilio polymnestor, AND Euploea core FROM JHARKHAND (INDIA) USING ADVANCED BIOTECHNOLOGICAL AND BIOINFORMATIC APPROACHES

Kumar Manoj ¹. Ranjan Rakesh ¹, Pratik Raj ¹, Tirkey Suraj ¹, Kumar Aman ¹, Hembrom Twinkle ¹, Prasad S. Manoranjan ², Singh R. Bharti¹

¹ St. Xavier’s College, Faculty of Science, Department of Zoology, Jharkhand, India
² Ranchi University, Faculty of Science, Department of Zoology, Jharkhand, India

Plants and butterflies have a coevolutionary relationship, and butterflies are essential to the ecosystem.
They serve as ecosystem indicators as well since research on their populations and behaviour can reveal how healthy an environment is. They are efficient pollinators, and particular species have been
known to migrate great distances to spread pollen, which causes genetic variety in plant species and increases their chances of surviving.

Aim. This work aimed to study the morpho-molecular features of Papilio polytes, Papilio polymnestor, and Euploea core from the Jharkhand state of India.

Methods. The study spans different areas from five districts, viz. Chatra, Koderma, Giridih, Godda, Ramgarh of the state. The specimens were collected, examined, and physical specimens of each species were submitted to the Insect Collection, Record and Identification, of the Department of Zoology, St. Xavier’s College, Ranchi, and Voucher numbers were obtained. Modern biotechnological and bioinformatics tools were used in this work, five specimens of each species were sequenced for the mitochondrial cytochrome oxidase subunit 1 (CO1), on the basis of which the BLAST (Basic Local Alignment and Search Tool) search was performed for identification of the species on the basis of matching scores with sequences present in the nucleotide sequence databases.

Results. The latest biotechnologies were used. After identification, the sequences were submitted to
GenBank, and accession IDs were obtained. The sequences were used to prepare a phylogenetic tree to ascertain the relationships among the collected specimens.

Conclusions. There is a paucity of knowledge related to the morphology and taxonomy of utterflies of the state; thus, this study is the first attempt of its kind. The study revealed significant ntraspecific variation among specimens of Euploea core. The least variation was exhibited among specimens of Papilio polymnestor. The study contributes to the knowledge related to butterfly species of Papilio polytes, Papilio polymnestor, and Euploea core. It will be helpful in further studies related to the conservation and monitoring of the species.

Key words: morpho-molecular, Jharkhand, India, Papilio polytes, Papilio polymnestor, Euploea core, BLAST, phylogeny.

References

Walker, B.H. (1995). Rangeland Ecology: Managing Change in Biodiversity. In: Perrings, C.A., Mäler, KG., Folke, C., Holling, C.S., Jansson, BO. (eds) Biodiversity Conservation. Ecology, Economy & Environment, v. 4. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-0277-3_5
Potts, S.G., Biesmeijer, J.C., Kremen, C., Neumann, P., Schweiger, O, Kunin, W.E. (2010). Global pollinator declines: Trends, impacts and drivers. Trends Ecol. Evol. 25, 345–353. https://doi.org/10.1016/j.tree.2010.01.007
Goulson, D., Nicholls, E, Botía, C., Rotheray, E.L. (2015). Bee declines driven by combined stress from parasites, pesticides, and lack of flowers. Science 347, 1255957. https://doi.org/10.1126/science.1255957
Bergholz, K., Sittel, L.P, Ristow, M., Jeltsch, F., Weiss, L. (2022). Pollinator guilds respond contrastingly at different scales to landscape parameters of land-use intensity. Ecol. Evol. 12, e8708. https://doi.org/10.1002/ece3.8708
Klein, A.M., Vaissiere, B.E., Cane, J.H., Steffan-Dewenter, I., Cunningham, S.A., Kremen, C., Tscharntke, T. (2007). Importance of pollinators in changing landscapes for world crops. R. Soc. B Biol. Sci. 274, 303–313. https://doi.org/10.1098/ rspb.2006.3721. https://doi.org/10.1098/rspb.2006.3721
Rader, R., Bartomeus, I., Garibaldi, L.A., Garratt, M.P.D., Howlett, B.G., Winfree, R.,….Woyciechowski, M. (2016). Non-bee insects are important contributors to global crop pollination. Natl. Acad. Sci. 113 (1), 146–151. https://doi.org/10.1073/pnas.1517092112
Gallai, N., Salles, J.M., Settele, J., Vaissiere, B.E. (2009). Economic valuation of the vulnerability of world agriculture confronted with pollinator decline. Econ. 68, 810–821. https://doi.org/10.1016/j.ecolecon.2008.06.014
Sharma, S., Mansotra, D.K., Joshi, P.C. (2020) Role of butterflies in shaping an ecosystem: why to protect them?. Ecology and Biodivdersity :39-44 Editor : Prof. P.C. Joshi, N. Joshi, B. N. Pandey, and D. K. Mansotra Today & Tomorrow’s Printers and Publishers, New Delhi - 110 002
Segre, H., David, K., Ignasi, B., Michiel, F., Mark, J., Maarten, F.S., Jacinto, R., Thijs, P.M.F. (2023). Butterflies are not a robust bioindicator for assessing pollinator communities, but floral resources offer a promising way forward. Ecological Indicators. 154 (2023) 110842. https://doi.org/10.1016/j.ecolind.2023.110842
O’Connor, R.S., Kunin, W.E., Garratt, M.P.D., Potts, S.G., Roy, H.E., Andrews, ….Carvalheiro, L. (2019). Monitoring insect pollinators and flower visitation: The effectiveness and feasibility of different survey methods. Methods Ecol. Evol. 10 (12), 2129–2140. https://doi.org/10.1111/2041-210X.13292
Mandelik, Y., Roll, U., Fleischer, A. (2010). Cost-efficiencyof biodiversity indicators for Mediterranean ecosystems and the effects of socio-economic factors. J. Appl. Ecol. 47, 1179–1188. https://doi.org/10.1111/j.1365-2664.2010.01864.x .
Breeze, T.D., Alison, P.B., Kelvin, G.B., Tom, B., …. Claire, C. (2020). Pollinator monitoring more than pays for itself. Applied Ecology. https://doi.org/10.1111/1365-2664.13755
Singh, N., Ahmad, J. (2017). A preliminary list of lepidopteran insects from Palkot Wildlife Sanctuary, Jharkhand. Journal of Entomology and Zoology Studies. 5(3), 654-661.
Kumari, K., Oraon, P.K., Kumari, K., Kumari, A., Banra, S., Thakur, A.K. (2021). Inventory report of butterflies (Lepidoptera: Rhopalocera) of family Nymphalidae, Pieridae and Papilionidae at Kanke, Ranchi. Biospectra, 16(2). 169 – 174.
Kumari, K., Thakur, A.K. (2022). Studies on the biodiversity of some families of butterflies (Lepidoptera: Rhopalocera) at Kanke, Ranchi, Jharkhand, India. Biospectra. 17(1), 93-98.
Kumar, M., Ranjan, R., Kumar, A., Pratik, R., Hembrom, T., Tirkey, S. (2025). Current studies on butterfly conservation in Jharkhand: A Data-Oriented Analysis. Vat-Vriksha. 1(1): OM03. https://doi.org/10.5281/zenodo.15834007 https://doi.org/10.5281/zenodo.12533671
Mahto, D., Osga, J., Khan, M.A., Kunkal, R., Herenj, S., Minz, P.A., Pratap, R., Mahto, S., Masarat, F. (2023). Diversity and conservation of Rhopalocera in urban Ranchi. In Jeune Chercheur (Volume II): etudes en Entomologie. Ed. Manoj Kumar, Notion Press (ISBN: 9798890265234)
Osga, J., Kumar, M., Mahto, D., Minz, P.A., Herenj, S., Kunkal, R.L, Mahto, S., Khan, M., …., Raipat, B.S. (2025). Butterflies from the St. Xavier’s College (Ranchi) campus: A Checklist. Vat-Vriksha. 1(1): OM06. https://doi.org/10.5281/zenodo.16572855 https://doi.org/10.2139/ssrn.5399209
Kunte, K. 2006. India-A lifescape: Butterflies of Peninsular India. ISBN: 8173713545. Universities Press. Hyderabad, India.
Smetacek, P. (2022). A naturalist’s guide to the Butterflies of India, Bangladesh, Nepal, Pakistan and Sri Lanka. ISBN978-81-7599-406-5. John Beaufoy Publishing Ltd.
Kumar, M., Ranjan, R., Sinha, M.P., Dhan, A., Naaz, F., Khanum, G., Rani, K.A., Sharma, S., Raipat, B.S. (2022). A review on insect collection and preservation techniques. European Journal of Pharmaceutical and Medical Research, 9(7): 233-239. https://papers.ssrn.com/sol3/papers.cfm?abstract_id=4171417
Mehra, D., Kirti, J.S., Sidhu, A.K. (2018). A taxonomic study of six species of the genus Junonia Hubner, [1819] (Insecta: Lepidoptera: Nymphalidae) from the northwestern Himalayan region in India. Journal of Threatened Taxa. 10(7), 11934 – 11947. https://doi.org/10.11609/jott.3731.10.7.11934-11947
ThermoFisher (2024): Applied Biosystems 3500 Dx Series Genetic Analyzer (CE-IVD, IVDR). Retrieved on 25^th May, 2024; 08, 02 (IST). url: https://www.thermofisher.com/ie/en/home/life-science/sequencing/sanger-sequencing/sanger-sequencing-technology-accessories/applied-biosystems-sanger-sequencing-3500-series-genetic-analyzers/3500-series-genetic-analyzer/3500-dx-cs2-series-genetic-analyzer.html
Srivastava, R., Kumar, M., Ranjan, R., Dandapat, S., Sinha, M.P. (2024). Morpho-molecular taxonomy and bioinformatics based phylogeny of Metaphire posthuma (Sims and Easton, 1972). The Bioscan. 19(1), 45-52. DOI: https://dx.doi.org/10.2139/ssrn.4847732
Tamura, K., Nei, M., Kumar, S. (2004). Prospects for inferring very large phylogenies by using the neighbor-joining method. Proceedings of the National Academy of Sciences (USA). 101, 11030-11035. DOI: https://doi.org/10.1073/pnas.0404206101
Kimura, M. (1980). A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution. 16, 111 – 120. https://doi.org/10.1007/BF01731581
Kumar, S., Stecher, G., Li, M., Knyaz, C., Tamura, K. MEGA X: Molecular Evolutionary Genetics Analysis across computing platforms. Molecular Biology and Evolution35, 1547-1549. DOI: https://doi.org/10.1093/molbev/msy096
Manoj, K., Anshu, P., Rakesh, R., Manoranjan, P.S., Bharti, S.R. (2024). Morphological study and DNA barcoding of Junonia atlites collected from Jharkhand state of India. International Journal of Entomology Research, 9(11), 1 – 7. https://dx.doi.org/10.2139/ssrn.5096390
Rajeswari, N.B., Jeyabalan, D. (2017). Studies on biology and reproduction of butterflies (family: Papilionidae) in Nilgiri Hills, Southern Western Ghats, India. International Journal of Advanced Research in Biological Sciences. 4(2), 1 – 11. http://dx.doi.org/10.22192/ijarbs.2017.04.02.001
Layberry, R.A., Hall, P.W., Lafontaine, J.D. (1998). The butterflies of Canada. University of Toronto Press, Toronto. https://doi.org/10.3138/9781442623163
Kioko, E., Musyoki, A.M., Kioko, M.D., Mwangi, E.W., Monda, L (2020). Swallowtail butterflies (Lepidoptera: Papilionidae) species diversity and distribution in Africa: The Papilionidae collection at the National Museums of Kenya, Nairobi, Kenya. Biodiversity Data Journal 8:e50664. https://doi.org/10.3897/BDJ.8.e50664
Kunte, K. 2013. "Research: Molecular genetics of mimicry in the Papilio polytesbutterfly" (On-line). Biodiversity Lab. Accessed May 28, 2013 at http://biodiversitylab.org/index/index.php/research/84-research/84-Papilio-polytes-mimicry-genetics.
Rangana, N. (2010). "Common Mormon" (On-line). Butterflies of Sri Lanka. Accessed February 10, 2012 at http://www.wildreach.com/butterflies/Papilio_polytes.php.
Kumar, M., Sahu, D., Kujur, A., Ehsan, H., Hembrom, L., Sinha, N., …. Raipat, B.S. (2024). First distributional record of the rare blue Mormon (Papilio polymnestor) from Gumla district (Jharkhand, India). European Journal of Pharmaceutical and Medical Research, 11(6), 195 – 196. https://doi.org/10.5281/zenodo.15528085
Antil, S., Abraham, J.S., Sripoorna, S., Maurya, S., Dagar, J., Makhija, S., Bhagat, P., Gupta, R., Sood, U., Lal, R., Toteja, R. (2022). DNA barcoding, an effective tool for species identification: a review. Wiley Oneline Library. 50(1), 761-775. https://doi.org/10.1155/2014/406178
Kress, W.J., Erickson, D.L. (2008). DNA barcodes: Genes, genomics, and bioinformatics. PNAS 105(8), 2761-2762. https://doi.org/10.1073/pnas.0800476105
Mampang, R.T., Auxtero, K.C.A., Caldito, C.J.C., Abanilla JM, Santos GAG, Caipang CMA (2023). DNA barcoding and its applications: A Review. Uttar Pradesh Journal of Zoology, 44(20), 69-78. https://doi.org/10.56557/upjoz/2023/v44i203646
Zheng, Z., Scott, S., Lukas, W., Webb, M. (2000). A greedy algorithm for aligning DNA sequences. Comput. Biol., 7(1-2), 203 – 214. https://doi.org/10.1089/10665270050081478
Aleksandr, M., George, C., Yan, R., Thomas, L.M., Richa, A., Schaffer, A.A. (2008). Database indexing for production MegaBLAST searches. Bioinformatic, 24, 1757-1764. https://doi.org/10.1093/bioinformatics/btn322
Kloczkowski, A., Jernigan, R.L., Wu, Z., Song, G., Yang, L., Kolinski, A., Pokarowski, P. (2011). Distance matrix-based approach to protein structure prediction. Journal of Struct. Funct. Genomics. 10(01), 67-81. https://doi.org/10.1007/s10969-009-9062-2
Al-Neama, M., Reda, N.M., Ghaleb, F.F.M. (2014). An improved distance matrix computation algorithm for multicore clusters. Res. Int. 2014, 406178. https://doi.org/10.1155/2014/406178
Baum, D. (2008). Reading a Phylogenetic tree: Meaning of monophyletic groups. Nature Education 1(1), 190. https://www.nature.com/scitable/topicpage/reading-a-phylogenetic-tree-the-meaning-of-41956/
Saitou, N., Nei, M. (1987). The neighbor-joining method: A new method for reconstructing phylogenetic trees. Molecular Biology and Evolution. 4. 406-425. https://doi.org/10.1093/oxfordjournals.molbev.a040454
Felsenstein, J. (1985). Confidence limits on phylogenies: An approach using the bootstrap. Evolution 39,:783-791. https://doi.org/10.2307/2408678