Molecular characterization of the grasshopper fauna in the Malakand region, Pakistan

Abdul Aziz, Munawar Saleem Ahmad, Kausar Saeed, Israr Alam, Saddam Hussain, Fawad Khan

Abstract


The first systematic inventory of grasshoppers (Orthoptera: Acrididae) in the Malakand Division of Pakistan was conducted from April 2022 to April 2023. A total of 2,395 specimens were collected from 26 sites across eight districts using standardized sweep-netting and hand-collection methods, followed by morphometric identification based on total body length (TBL), pronotal length (PL), abdominal length (AL), tegmen length (TL), and hind femur length (HFL). Thirty-one species were recorded, representing 22 genera, 15 tribes, and nine subfamilies of Caelifera. Acridinae, Oedipodinae, Cyrtacanthacridinae, and Gomphocerinae were the most diverse subfamilies. Species richness and encounter frequency were positively correlated with warm, dry conditions, peaking in mixed-cropping zones of Swat, Buner, and Bajaur. Morphometric data revealed significant size variation, with Choroedocus robustus (TBL 60 mm, HFL 33 mm) and C. illustris (TBL 52 mm, HFL 40 mm) as the largest species, contrasting with smaller taxa such as Phlaeoba panteli and Aulacobothrus socius (TBL 13-14 mm), reflecting habitat adaptations and escape behavior. COXI DNA barcoding confirmed most morphological identifications. The nucleotide composition showed an AT bias (≈60-68%), R-values ranged from 1.02 to 3.87, and interspecific genetic distances (d) varied from 0.01 to 0.37. Pairwise divergence among Acrida, Phlaeoba, and Truxalis indicated genetic coherence, whereas higher distances in Leva indica and Crucinotacris decisa suggested broader geographic variation. Phylogenetic analysis using the Kimura 2-parameter model supported subfamily-level clades. Ecologically, assemblages included pest species such as Cyrtacanthacris tatarica and beneficial taxa vital for nutrient cycling, underscoring the need for context-specific conservation strategies.

Keywords


Grasshopper diversity, Molecular characterization, DNA barcoding, Insect taxonomy, Pakistan fauna

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Akhtar, N., Tahir, H.M., Azizullah, A., Ali, A., Fajar, R., Muzamil, A., Abbas, D., 2024. Survey and seasonal abundance of major insect pests in the maize fields of Punjab, Pakistan. International Journal of Tropical Insect Science 45(5), 2211-2224.

Ali, F., Khan, N., Ahmad, A., Khan, A.A., 2019. Structure and biomass carbon of Olea ferruginea forests in the foot hills of Malakand division, Hindukush Range Mountains of Pakistan. Acta Ecologica Sinica 39(4), 261-266.

Ali, S., Ali, H., Baharanchi, O.G., Sher, H., Yousefpour, R., 2024. Investigating endemic species conservation hotspots based on species distribution models in Swat region, Hindu Kush Pakistan. Land 13(6), 737.

Barandica, J.C.N., Quintero-Galvis, J.F., Vargas, A.C., Cervantes, G.B., Pabón, J.C.A., García, L.C., Acero, A., 2024. Congruence between microsatellite loci, SNPs, and COI gene reveals a phylogeographic break in the southern Caribbean Sea: The case of West Indian top shell Cittarium pica (Gastropoda: Tegulidae). Fisheries Research 269, 106857.

Barandica, R., Garimella, R., Letsiou, T., 2024. Molecular diversity and species identification of grasshoppers through mitochondrial DNA barcoding: Insights into the evolutionary patterns of Acrididae. Journal of Molecular Entomology 56(2), 102-119.

Blanco-Sierra, L., Bellver-Arnau, J., Escartin, S., Mariani, S., Bartumeus, F., 2024. Human-environment interactions shape mosquito seasonal population dynamics. Insects 15(7), 527.

Chapco, W., Contreras, D., 2011. Subfamilies Acridinae, Gomphocerinae and Oedipodinae are “fuzzy sets”: a proposal for a common African origin. Journal of Orthoptera Research 20(2), 173-190.

Chaudhary, A., Chauhan, P., Singh, A.K., Rao, I., 2025. Phylogenetic analysis of Steinernema pakistanense from the semi-arid regions of India and its toxicity assessment against lepidopteran and coleopteran insects. Egyptian Journal of Biological Pest Control 35(1), 14.

Chen, L.-P., Chang, Y.-G., Li, J., Wang, J.-M., Liu, J.-L., Zhi, Y.-C., Li, X.J., 2018. Application of DNA barcoding in the classification of grasshoppers (Orthoptera: Acridoidea)-A case study of grasshoppers from Hebei province, China. Zootaxa 4497(1), 99-110.

Das, S.R., Islam, M.I., Aslam, A.F.M., 2022. Molecular characterization and identification of grasshopper pests of rice inferred from mitochondrial COI gene sequences. Bangladesh Journal of Zoology 50(2), 217-229.

Du, B., Wei, J., Lin, K., Lu, L., Ding, X., Ye, H., Wang, N., 2022. Spatial and temporal variability of grassland grasshopper habitat suitability and its main influencing factors. Remote Sensing 14(16), 3910.

Garimella, S.S.S., Rachakonda, S.V., Pratapa, S.S., Mannem, G.D. Mahidhara, G., 2024. From cells to power cells: harnessing bacterial electron transport for microbial fuel cells (MFCs). Annals of Microbiology 74(1), 19.

Guzmán, N.V., Gandini, L.M., Castillo, E.R., Fernandez Campón, F., Cigliano, M.M., Confalonieri, V.A., 2024. Pleistocene climatic oscillations influenced the emergence of geographically widespread and restricted genetic lineages in an Andean grasshopper species group. Biological Journal of the Linnean Society 143(1), blad166.

Hussain, M., Kazam, S., Noreen, A., Shah, S., Zeb, U., Iftikhar, A., 2024. Diversity and distributional patterns of grasshoppers in croplands of district Gujrat, Punjab, Pakistan. Biologia (Lahore) 70(2), 38-46.

Jannat, R., Shaha, M., Rubayet, M.T., Sultana, S., 2018. Role of chitosan in induction of defense response against Phomopsis vexans and augmentation of growth and yield of eggplant. Global Journal of Science Frontier Research: C Biological Science 18(3), 6-13.

Jayaprasad, S., Peona, V., Ellerstrand, S.J., Rossini, R., Bunikis, I., Pettersson, O.V., Olsen, R.A., Rubin, C.J., Einarsdottir, E., Bonath, F., Bradford, T.M., 2024. Orthopteran neo‐sex chromosomes reveal dynamics of recombination suppression and evolution of supergenes. Molecular Ecology 33(23), e17567.

Kalhoro, A.S., Baloch, N., Soomro, I.A., Memon, A., 2019. Grasshopper fauna (acrididae) of three zones of District Dadu, Sindh, Pakistan. Journal of Entomology & Zoology Studies 7, 49-51.

Karso, B.A., 2022. Bio effect of seven aqueous plant extracts against khapra beetle Trogoderma Granarium Everts. NTU Journal of Agriculture and Veterinary Science 2(1), 9-13.

Khalil, A., Rafiq, N., Khalil, A., Haider, L., 2025. Comparative study on grasshopper diversity and desert locust in wet and dry soil areas of Kuchlak (Quetta District) of Balochistan. Pak-Euro Journal of Medical and Life Sciences 8(1), 101-106.

Khan, Z., Ikhtiar, Z., Rahman, A.S.W., Rahman, H.U, Mehmood, I.A., Alam, A., Khan, F., 2021. 22. Diversity of grasshopper in the piedmont of mount Elum, Buner. Pure and Applied Biology (PAB) 11(1), 217-225.

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.

Kumar, S., Stecher, G., Tamura, K., 2016. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33(7), 1870-1874.

Letsiou, S., Madesis, P., Vasdekis, E., Montemurro, C., Grigoriou, M.E., Skavdis, G., Tzakos, A.G., 2024. DNA barcoding as a plant identification method. Applied Sciences 14(4), 1415.

Mustafa, S.B., Panhwar, W.A., Shaikh, A.M., 2024. Seasonal distribution and systematic study of Grasshopper (Acrididae) family: A regional study of Tehsil Ratodero, Sindh Pakistan. Journal of Entomology and Zoology Studies 12(2), 12-16.

Panhwar, W.A., Ahmed, S., Mehmood, S.A., Shaikh, A.M., Khan, W., Shah, M., 2024. Diversity, species richness and distribution of grasshoppers (Orthoptera: Acrididae) in Khairpur Sindh Pakistan. International Journal of Tropical Insect Science 44(5), 2501-2510.

Pitafi, M.R., 2024. Integrating statistical models and principal component analysis to assess grasshopper distribution and diversity in Matiari district, Sindh, Pakistan. Journal of Advanced Zoology 45(6).

Povea-Cabello, S., Brischigliaro, M., Fernández-Vizarra, E., 2024. Emerging mechanisms in the redox regulation of mitochondrial cytochrome c oxidase assembly and function. Biochemical Society Transactions 52(2), 873-885.

Ramakanta Rao, V., 1984. Taxonomy and behavior of Acrididae: Grasshoppers and locusts. Journal of Entomology 12(4), 30-45.

Riffat, S., Wagan, M., 2015. Grasshoppers and locusts of Pakistan. Higher Education Commission, Pakistan 1-180.

Riyaz, A., 2021. Grasshoppers and the order Orthoptera: Taxonomy and classification. Journal of Entomological Studies, 15(3), 12-19.

Sarwar, Z., Tamkeen, A., Khan, M., Hussain, Z., Asghar, M., Badar, M., 2023. Spatiotemporal behavior and new distributional record of subfamily hemiacridinae (Acrididae: Orthoptera) from Azad Jammu and Kashmir, Pakistan. Journal of Animal & Plant Sciences 33(3).

Sayers, E.W., Cavanaugh, M., Clark, K., Pruitt, K.D., Sherry, S.T., Yankie, L., Karsch-Mizrachi, I., 2024. GenBank 2024 update. Nucleic Acids Research 52(D1), D134-D137.

Singh, N., Kumari, V., 2021. Locusts. Polyphagous Pests of Crops, 1-50.

Sukhikh, I., Bugrov, A., Chobanov, D., Fet, V., Biryukov, M., Blinov, A., 2025. Molecular phylogenetic analysis and taxonomy of the Palearctic Pamphagidae grasshoppers (Orthoptera, Acridoidea) based on DNA markers. Insect Systematics & Evolution 56(3), 235-247.

Ullah, I., Akhtar, K.M., Shahzadi, I., Farooq, M., Yasmin, R., 2016. Encouraging knowledge sharing behavior through team innovation climate, altruistic intention and organizational culture. Knowledge Management & E-Learning 8(4), 631.

Wang, H., Jiang, B., Gu, J., Wei, T., Lin, L., Huang, Y., Huang, J., 2021. Molecular phylogeny and species delimitation of the genus Tonkinacris (Orthoptera, Acrididae, Melanoplinae) from China. Plos One 16(4), e0249431.

Waqar, Z., Ahmed, S., Mehmood, S.A., Panhwar, W.A., Ahmad, B., Khan, S., 2024. Diversity and distributional pattern of grasshoppers fauna in agroforest and vegetative lands of tehsil Havelian, Pakistan.

Zahid, S., Mariño-Pérez, R., Song, H., 2021. Molecular phylogeny of the grasshopper family Pyrgomorphidae (Caelifera, Orthoptera) reveals rampant paraphyly and convergence of traditionally used taxonomic characters. Zootaxa 4969(1), 101-118.

Zhang, D., Gao, F., Jakovlić, I., Zou, H., Zhang, J., Li, W.X., Wang, G.T., 2020. PhyloSuite: An integrated and scalable desktop platform for streamlined molecular sequence data management and evolutionary phylogenetics studies. Molecular Ecology Resources 20(1), 348-355.




DOI: https://doi.org/10.33804/pp.009.04.5890

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