Cryptic species-specific transmission of the invasive begomovirus tomato leaf curl New Delhi virus by the whitefly Bemisia tabaci

Abstract

Tomato leaf curl New Delhi virus (ToLCNDV), a bipartite begomovirus initially identified in tomato, has rapidly spread worldwide, emerging as a significant threat to cucurbit and other crops. Although geminiviruses are typically transmitted by the whitefly Bemisia tabaci, the specific cryptic species responsible for ToLCNDV transmission remains largely unclear. In this study, we collected diseased tomato and cucurbit plants, along with associated whiteflies, in China, and detected ToLCNDV in both plant and whitefly samples. We then compared the acquisition and transmission efficiency of ToLCNDV among whiteflies of three B. tabaci cryptic species: the invasive Middle East-Asia Minor 1 (MEAM1), Mediterranean (MED), and the indigenous Asia II 1. Our findings revealed that whiteflies of three species were capable of acquiring ToLCNDV; however, MED and Asia II 1 transmitted the virus with significantly higher efficiency than MEAM1. Tissue-specific viral load analysis further indicated that the virus crossed the midgut wall more effectively in MED and Asia II 1 compared to MEAM1. These results provide direct evidence that B. tabaci transmits ToLCNDV in a cryptic species-dependent manner. Given the global prevalence of B. tabaci, the potential role of specific cryptic species in driving the spread of ToLCNDV warrants close monitoring and proactive management strategies.

Keywords

Bemisia tabaci / cryptic species-dependent / tomato leaf curl New Delhi virus / virus transmission

References

[1] Padidam, M., Beachy, R. N., & Fauquet, C. M. (1995). Tomato leaf curl geminivirus from India has a bipartite genome and coat protein is not essential for infectivity. Journal of General Virology, 76(1), 25–35. https://doi.org/10.1099/0022-1317-76-1-25

[2] Srivastava, K. M., Hallan, V., Raizada, R. K., Chandra, G., Singh, B. P., & Sane, P. V. (1995). Molecular cloning of Indian tomato leaf curl virus genome following a simple method of concentrating the supercoiled replicative form of viral DNA. Journal of Virological Methods, 51(2), 297–304. https://doi.org/10.1016/0166-0934(94)00122-W

[3] Zaidi, S. S. E. A., Martin, D. P., Amin, I., Farooq, M., & Mansoor, S. (2017). Tomato leaf curl New Delhi virus: A widespread bipartite begomovirus in the territory of monopartite begomoviruses. Molecular Plant Pathology, 18(7), 901–911. https://doi.org/10.1111/mpp.12481

[4] Cai, L., Mei, Y., Ye, R., Deng, Y., Zhang, X., Hu, Z., Zhou, X., Zhang, M., & Yang, J. (2023). Tomato leaf curl New Delhi virus: An emerging plant begomovirus threatening cucurbit production. aBIOTECH, 4(3), 257–266. https://doi.org/10.1007/s42994-023-00118-4

[5] Siskos, L., Cui, L., Wang, C., Visser, R. G. F., Bai, Y., & Schouten, H. J. (2022). A new challenge in melon resistance breeding: The ToLCNDV case. Euphytica, 218(9), 129. https://doi.org/10.1007/s10681-022-03081-1

[6] Venkataravanappa, V., Ashwathappa, K. V., Reddy, C. N. L., Shankarappa, K. S., & Reddy, M. K. (2020). Characterization of tomato leaf curl New Delhi virus associated with leaf curl and yellowing disease of watermelon and development of LAMP assay for its detection. 3 Biotech, 10(6), 282. https://doi.org/10.1007/s13205-020-02245-x

[7] Yamamoto, H., Wakita, Y., Kitaoka, T., Fujishiro, K., Kesumawati, E., & Koeda, S. (2021). Southeast Asian isolate of the tomato leaf curl New Delhi virus shows higher pathogenicity against tomato and cucurbit crops compared to that of the Mediterranean isolate. Horticulture Journal, 90(3), 314–325. https://doi.org/10.2503/hortj.UTD-269

[8] Fiallo-Olivé, E., Pan, L. L., Liu, S. S., & Navas-Castillo, J. (2020). Transmission of begomoviruses and other whitefly-borne viruses: Dependence on the vector species. Phytopathology, 110(1), 10–17. https://doi.org/10.1094/PHYTO-07-19-0273-FI

[9] Wang, X. W., & Blanc, S. (2021). Insect transmission of plant single-stranded DNA viruses. Annual Review of Entomology, 66(1), 389–405. https://doi.org/10.1146/annurev-ento-060920-094531

[10] Ghanim, M., Morin, S., & Czosnek, H. (2001). Rate of tomato yellow leaf curl virus translocation in the circulative transmission pathway of its vector, the whitefly Bemisia tabaci. Phytopathology, 91(2), 188–196. https://doi.org/10.1094/PHYTO.2001.91.2.188

[11] Wei, J., Zhao, J. J., Zhang, T., Li, F. F., Ghanim, M., Zhou, X. P., Ye, G. Y., Liu, S. S., & Wang, X. W. (2014). Specific cells in the primary salivary glands of the whitefly Bemisia tabaci control retention and transmission of begomoviruses. Journal of Virology, 88(22), 13460–13468. https://doi.org/10.1128/jvi.02179-14

[12] Czosnek, H., Hariton-Shalev, A., Sobol, I., Gorovits, R., & Ghanim, M. (2017). The incredible journey of begomoviruses in their whitefly vector. Viruses, 9(10), 273. https://doi.org/10.3390/v9100273

[13] Rosen, R., Kanakala, S., Kliot, A., Cathrin Pakkianathan, B., Farich, B. A., Santana-Magal, N., Elimelech, M., Kontsedalov, S., Lebedev, G., Cilia, M., & Ghanim, M. (2015). Persistent, circulative transmission of begomoviruses by whitefly vectors. Current Opinion in Virology, 15, 1–8. https://doi.org/10.1016/j.coviro.2015.06.008

[14] Pan, L., Chen, Q., Guo, T., Wang, X., Li, P., Wang, X., & Liu, S. (2018). Differential efficiency of a begomovirus to cross the midgut of different species of whiteflies results in variation of virus transmission by the vectors. Science China Life Sciences, 61(10), 1254–1265. https://doi.org/10.1007/s11427-017-9283-4

[15] Guo, T., Zhao, J., Pan, L. L., Geng, L., Lei, T., Wang, X. W., & Liu, S. S. (2018). The level of midgut penetration of two begomoviruses affects their acquisition and transmission by two species of Bemisia tabaci. Virology, 515, 66–73. https://doi.org/10.1016/j.virol.2017.12.004

[16] Wang, H. L., Lei, T., Wang, X. W., Cameron, S., Navas-Castillo, J., Liu, Y. Q., Maruthi, M. N., Omongo, C. A., Delatte, H., Lee, K. Y., Krause-Sakate, R., Ng, J., Seal, S., Fiallo-Olivé, E., Bushley, K., Colvin, J., & Liu, S. (2025). A comprehensive framework for the delimitation of species within the Bemisia tabaci cryptic complex, a global pest-species group. Insect Science, 32(1), 321–342. https://doi.org/10.1111/1744-7917.13361

[17] Kil, E. J., Vo, T. T. B., Fadhila, C., Ho, P. T., Lal, A., Troiano, E., Parrella, G., & Lee, S. (2020). Seed transmission of tomato leaf curl New Delhi virus from zucchini squash in Italy. Plants, 9(5), 563. https://doi.org/10.3390/plants9050563

[18] Fortes, I. M., Perez-Padilla, V., Romero-Rodriguez, B., Fernandez-Munoz, R., Moyano, C., Castillo, A. G., De Leon, L., & Moriones, E. (2023). Begomovirus tomato leaf curl New Delhi virus is seedborne but not seed transmitted in melon. Plant Disease, 107(2), 473–479. https://doi.org/10.1094/PDIS-09-21-1930-RE

[19] López, C., Ferriol, M., & Picó, M. B. (2015). Mechanical transmission of tomato leaf curl New Delhi virus to cucurbit germplasm: Selection of tolerance sources in Cucumis melo. Euphytica, 204(3), 679–691. https://doi.org/10.1007/s10681-015-1371-x

[20] Janssen, D., Simón, A., Boulares, M., & Ruiz, L. (2022). Host species-dependent transmission of tomato leaf curl New Delhi virus-ES by Bemisia tabaci. Plants, 11(3), 390. https://doi.org/10.3390/plants11030390

[21] Sivagnanapazham, K., Karthikeyan, G., Pavithran, S., Harish, S., Murugan, M., Latha, T. K. S., & Devi, H. U. N. (2025). Transmission dynamics and localization of tomato leaf curl New Delhi virus in cucurbits via sweet potato whitefly, Bemisia tabaci (Gennadius). Molecular Biology Reports, 52(1), 358. https://doi.org/10.1007/s11033-025-10467-6

[22] Shan, H., Liu, Y., Luan, J., & Liu, S. (2021). New insights into the transovarial transmission of the symbiont Rickettsia in whiteflies. Science China Life Sciences, 64(7), 1174–1186. https://doi.org/10.1007/s11427-020-1801-7

[23] Vignesh, S., Renukadevi, P., Nagendran, K., Senthil, N., Kumar, R. V., SwarnaPriya, R., Behera, T. K., & Karthikeyan, G. (2023). A distinct strain of tomato leaf curl New Delhi virus that causes mosaic disease in ash gourd and other cucurbitaceous crops. Frontiers in Microbiology, 14, 1268333. https://doi.org/10.3389/fmicb.2023.1268333

[24] Sinisterra, X. H., McKenzie, C. L., Hunter, W. B., Powell, C. A., & Shatters, R. G., Jr. (2005). Differential transcriptional activity of plant-pathogenic begomoviruses in their whitefly vector (Bemisia tabaci, Gennadius: Hemiptera Aleyrodidae). Journal of General Virology, 86(5), 1525–1532. https://doi.org/10.1099/vir.0.80665-0

[25] Sandra, N., & Mandal, B. (2024). Emerging evidence of seed transmission of begomoviruses: Implications in global circulation and disease outbreak. Frontiers in Plant Science, 15, 1376284. https://doi.org/10.3389/fpls.2024.1376284

[26] Lee, C. H., Zheng, Y. X., Chan, C. H., Ku, H. M., Chang, C. J., & Jan, F. J. (2020). A single amino acid substitution in the movement protein enables the mechanical transmission of a geminivirus. Molecular Plant Pathology, 21(4), 571–588. https://doi.org/10.1111/mpp.12917

[27] Teofanova, D., Lozanova, Y., Lambovska, K., Pachedjieva, K., Tosheva, A., Odjakova, M., & Zagorchev, L. (2022). Cuscuta spp. populations as potential reservoirs and vectors of four plant viruses. Phytoparasitica, 50(3), 555–566. https://doi.org/10.1007/s12600-022-00981-9

[28] Jia, D., Chen, Q., Mao, Q., Zhang, X., Wu, W., Chen, H., Yu, X., Wang, Z., & Wei, T. (2018). Vector mediated transmission of persistently transmitted plant viruses. Current Opinion in Virology, 28, 127–132. https://doi.org/10.1016/j.coviro.2017.12.004

[29] Pan, L. L., Cui, X. Y., Chen, Q. F., Wang, X. W., & Liu, S. S. (2018). Cotton leaf curl disease: Which whitefly is the vector? Phytopathology, 108(10), 1172–1183. https://doi.org/10.1094/PHYTO-01-18-0015-R

[30] Chen, T., Jia, Y., Tang, Y., Chen, J., Xu, H., & Qi, G. (2025). Cotton leaf curl Multan virus activates autophagy in the whitefly Asia II 7, weakening its vectorial capacity for transmission. Pest Management Science, 81(6), 3039–3047. https://doi.org/10.1002/ps.8674

[31] Idris, A. M., Smith, S. E., & Brown, J. K. (2001). Ingestion, transmission, and persistence of Chino del tomate virus (CdTV), a New World begomovirus, by Old and New World biotypes of the whitefly vector Bemisia tabaci. Annals of Applied Biology, 139(1), 145–154. https://doi.org/10.1111/j.1744-7348.2001.tb00139.x

[32] Pan, L. L., Chi, Y., Liu, C., Fan, Y. Y., & Liu, S. S. (2020). Mutations in the coat protein of a begomovirus result in altered transmission by different species of whitefly vectors. Virus Evolution, 6(1), veaa014. https://doi.org/10.1093/ve/veaa014

[33] Pan, L. L., Chen, Q. F., Zhao, J. J., Guo, T., Wang, X. W., Hariton-Shalev, A., Czosnek, H., & Liu, S. S. (2017). Clathrin-mediated endocytosis is involved in Tomato yellow leaf curl virus transport across the midgut barrier of its whitefly vector. Virology, 502, 152–159. https://doi.org/10.1016/j.virol.2016.12.029

[34] Ohnishi, J., Kitamura, T., Terami, F., & Honda, K. I. (2009). A selective barrier in the midgut epithelial cell membrane of the nonvector whitefly Trialeurodes vaporariorum to tomato yellow leaf curl virus uptake. Journal of General Plant Pathology, 75(2), 131–139. https://doi.org/10.1007/s10327-009-0147-3

[35] Zhao, J., Lei, T., Zhang, X. J., Yin, T. Y., Wang, X. W., & Liu, S. S. (2020). A vector whitefly endocytic receptor facilitates the entry of begomoviruses into its midgut cells via binding to virion capsid proteins. PLoS Pathogens, 16(12), e1009053. https://doi.org/10.1371/journal.ppat.1009053

[36] Rana, V. S., Popli, S., Saurav, G. K., Raina, H. S., Chaubey, R., Ramamurthy, V. V., & Rajagopal, R. (2016). A Bemisia tabaci midgut protein interacts with begomoviruses and plays a role in virus transmission. Cellular Microbiology, 18(5), 663–678. https://doi.org/10.1111/cmi.12538

[37] He, H. F., Zhao, C. C., Zhu, C. Q., Yan, W. L., Yan, M. H., Zhang, Z. L., Liu, J. L., Shi, B. Z., Bai, R. E., Li, J. J., & Yan, F. M. (2023). Discovery of novel whitefly vector proteins that interact with a virus capsid component mediating virion retention and transmission. International Journal of Biological Macromolecules, 226, 1154–1165. https://doi.org/10.1016/j.ijbiomac.2022.11.229

[38] Farooq, T., Lin, Q., She, X., Chen, T., Li, Z., Yu, L., Lan, G., Tang, Y., & He, Z. (2022). Cotton leaf curl Multan virus differentially regulates innate antiviral immunity of whitefly (Bemisia tabaci) vector to promote cryptic species-dependent virus acquisition. Frontiers in Plant Science, 13, 1040547. https://doi.org/10.3389/fpls.2022.1040547

[39] Wang, L. L., Wang, X. R., Wei, X. M., Huang, H., Wu, J. X., Chen, X. X., Liu, S. S., & Wang, X. W. (2016). The autophagy pathway participates in resistance to tomato yellow leaf curl virus infection in whiteflies. Autophagy, 12(9), 1560–1574. https://doi.org/10.1080/15548627.2016.1192749

[40] Wang, Z. Z., Bing, X. L., Liu, S. S., & Chen, X. X. (2017). RNA interference of an antimicrobial peptide, Btdef, reduces tomato yellow leaf curl China virus accumulation in the whitefly Bemisia tabaci. Pest Management Science, 73(7), 1421–1427. https://doi.org/10.1002/ps.4472

[41] Pinheiro, P. V., Kliot, A., Ghanim, M., & Cilia, M. (2015). Is there a role for symbiotic bacteria in plant virus transmission by insects? Current Opinion in Insect Science, 8, 69–78. https://doi.org/10.1016/j.cois.2015.01.010

[42] Kliot, A., Cilia, M., Czosnek, H., & Ghanim, M. (2014). Implication of the bacterial endosymbiont Rickettsia spp. in interactions of the whitefly Bemisia tabaci with tomato yellow leaf curl virus. Journal of Virology, 88(10), 5652–5660. https://doi.org/10.1128/jvi.00071-14

[43] Janssen, D., Simon, A., Crespo, O., & Ruiz, L. (2017). Genetic population structure of Bemisia tabaci in Spain associated with tomato leaf curl New Delhi virus – Short communication. Plant Protection Science, 53(1), 25–31. https://doi.org/10.17221/62/2016-PPS

[44] Moriones, E., Praveen, S., & Chakraborty, S. (2017). Tomato leaf curl New Delhi virus: An emerging virus complex threatening vegetable and fiber crops. Viruses, 9(10), 264. https://doi.org/10.3390/v9100264

[45] Juarez, M., Rabadan, M. P., Martinez, L. D., Tayahi, M., Grande-Perez, A., & Gomez, P. (2019). Natural hosts and genetic diversity of the emerging tomato leaf curl New Delhi virus in Spain. Frontiers in Microbiology, 10, 140. https://doi.org/10.3389/fmicb.2019.00140

[46] Islam, W., Akutse, K. S., Qasim, M., Khan, K. A., Ghramh, H. A., Idrees, A., & Latif, S. (2018). Bemisia tabaci-mediated facilitation in diversity of begomoviruses: Evidence from recent molecular studies. Microbial Pathogenesis, 123, 162–168. https://doi.org/10.1016/j.micpath.2018.07.008

[47] Lestari, S. M., Khatun, M. F., Acharya, R., Sharma, S. R., Shrestha, Y. K., Jahan, S. M. H., Aye, T. T., Lynn, O. M., Win, N. K. K., Hoat, T. X., Thi Dao, H., Tsai, C., Lee, J., Hwang, H., Kil, E., Lee, S., Kim, S., & Lee, K. (2023). Genetic diversity of cryptic species of Bemisia tabaci in Asia. Archives of Insect Biochemistry and Physiology, 112(2), e21981. https://doi.org/10.1002/arch.21981

[48] Parrella, G., Nappo, A. G., Manco, E., Greco, B., & Giorgini, M. (2014). Invasion of the Q2 mitochondrial variant of Mediterranean Bemisia tabaci in southern Italy: Possible role of bacterial endosymbionts. Pest Management Science, 70(10), 1514–1523. https://doi.org/10.1002/ps.3686

[49] Gauthier, N., Clouet, C., Perrakis, A., Kapantaidaki, D., Peterschmitt, M., & Tsagkarakou, A. (2014). Genetic structure of Bemisia tabaci Med populations from home-range countries, inferred by nuclear and cytoplasmic markers: Impact on the distribution of the insecticide resistance genes. Pest Management Science, 70(10), 1477–1491. https://doi.org/10.1002/ps.3733

[50] Henri, H., Terraz, G., Gnankiné, O., Fleury, F., & Mouton, L. (2013). Molecular characterization of genetic diversity within the Africa/Middle East/Asia Minor and Sub-Saharan African groups of the Bemisia tabaci species complex. International Journal of Pest Management, 59(4), 329–338. https://doi.org/10.1080/09670874.2013.869374

[51] Rehman, M., Chakraborty, P., Tanti, B., Mandal, B., & Ghosh, A. (2021). Occurrence of a new cryptic species of Bemisia tabaci (Hemiptera: Aleyrodidae): An updated record of cryptic diversity in India. Phytoparasitica, 49(5), 869–882. https://doi.org/10.1007/s12600-021-00909-9

RIGHTS & PERMISSIONS

2025 The Author(s). New Plant Protection published by John Wiley & Sons Australia, Ltd on behalf of Institute of Plant Protection, Chinese Academy of Agricultural Sciences.