Abstract
Tomato leaf curl New Delhi virus (ToLCNDV), a bipartite begomovirus, is the most important among the 14 species of begomoviruses infecting tomato in Indian subcontinent. Begomovirus is known to evade RNA silencing of host plants through suppressor proteins. However, in case of ToLCNDV, the suppressor proteins have not been studied well. The objective of the study is to know the sub-cellular localization of three suppressor proteins encoded by AV2, AC2 and AC4 ORFs of ToLCNDV in Nicotiana benthamiana. AV2, AC2 and AC4 ORFs of ToLCNDV were cloned and sequenced (accession numbers MW423574, MW423576, MW423575, respectively) from a ToLCNDV isolate characterized earlier (accession number MW429271) and GFP tagged constructs were prepared in a plant expressing binary vector pEarleygate103. Bioinformatics analysis using Peptide 2.0 server predicted that all these proteins have more basic amino acid residues then acidic amino acid and AV2 protein has more hydrophobic amino acid residues. ScanProsite server predicted presence of different fuctional motifs in these proteins amongst which presence of kinase motif was observed in all of them. Virus mPLoc server predicted their subcellular localization. The suppressor gene constructs were agroinfiltrated on to leaves of one month old N. benthamiana plants and their subcellular localization has been studied through confocal microscopy. Results have shown that AV2 localizes in the host cell membrane and nucleus, AC2 in the nucleus and AC4 in the host cell membrane. Earlier reports with other begomoviruses also showed similar localization behaviour of these suppressor protein except AV2, where it was shown to be present in cytoplasm. Such localization study will help understand the mechanism of their suppression activity.
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References
Malathi VG, Renukadevi P, Chakraborty S, Biswas KK, Roy A, Sivalingam PN, Venkataravanappa V, Mandal B. Begomoviruses and their satellites occurring in India: distribution, diversity and pathogenesis. A Century of Plant Virology in India, Springer, Singapore, 2017;pp. 75–177
Varma A, Mandal B, Singh MK. Globale and spread of whitefly (Bemisia tabaci) transmitted Geminiviruses. In: Thompson WMO, editor. 2011: The Whitefly, Bemisia tabaci (Homoptera: Aleyrodidae) Interaction with Geminivirus-Infected Host Plants. Springer: Netherlands; 2011. p. 205–92.
Shilpi S, Kumar A, Biswas S, Roy A, Mandal B. A recombinant Tobacco curly shoot virus causes leaf curl disease in tomato in a north-eastern state of India and has potentiality to trans-replicate a non-cognate betasatellite. Virus Genes. 2015;50:87–96.
Moriones E, Praveen S, Chakraborty S. Tomato leaf curl New Delhi virus: an emerging virus complex threatening vegetable and fiber crops. Viruses. 2007;9(10):264.
Dong X, van Wezel R, Stanley J, Hong Y. Functional characterization of the nuclear localization signal for a suppressor of post- transcriptional gene silencing. J Virol. 2003;77:7026–33.
van Wezel R, Liu H, Tien P, Stanly J, Hong Y. Gene C2 of the monopartite geminivirus Tomato yellow leaf curl virus-China encodes a pathogenicity determinant that is localized in the nucleus. Mol Plant- Microbe Interact. 2001;14:1125–8.
Chung HY, Lacatus G, Sunter G. Geminivirus AL2 protein induces expression of, and interacts with, a calmodulin-like gene, an endogenous regulator of gene silencing. Virology. 2014;460:108–18.
Roshan P, Kulshreshtha A, Hallan V. Identification of host cellular targets of AC4 and AV2 proteins of tomato leaf curl palampur virus and their sub-cellular localization studies. Virusdisease. 2017;28(4):390–400.
Doyle JJ, Doyle JL. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull. 1987;19:11–5.
Burgyan J, Havelda Z. Viral suppressors of RNA silencing. Trends Plant Sci. 2011;16:265–72.
Chowda-Reddy RV, Achenjang F, Felton C, Etarock MT, Anangfac MT, Nugent P, Fondong VN. Role of a geminivirus AV2 protein putative protein kinase C motif on subcellular localization and pathogenicity. Virus Res. 2008;135(1):115–24.
Shen W, Dallas MB, Goshe MB, Hanley-Bowdoin L. SnRK1 phosphorylation of AL2 delays Cabbage leaf curl virus infection in Arabidopsis. J Virol. 2014;88:10598–612.
Mei Y, Wang Y, Hu T, Yang X, Lozano-Duran R, Sunter G, Zhou X. Nucleocytoplasmic shuttling of the geminivirus C4protein mediated by phosphorylation and myristoylation is critical forviral pathogenicity. Mol Plant. 2018;11:1466–81.
Trinks D, Rajeswaran R, Shivaprasad PV, Akbergenov R, Oakeley EJ, Veluthambi K, Hohn T, Pooggin MM. Suppression of RNA silencing by a geminivirus nuclear protein, AC2, correlates with transactivation of host genes. J Virol. 2005;79:2517–27.
Carluccio AV, Prigigallo MI, Rosas-Diaz T, Lozano-Duran R, Stavolone L. S-acylation mediates Mungbean yellow mosaic virus AC4 localization to the plasma membrane and in turns gene silencing suppression. PLoS Pathog. 2018;14(8):e1007207.
Fondong VN, Reddy RC, Lu C, Hankoua B, Felton C, Czymmek K, Achenjang F. The consensus N-myristoylation motif of a geminivirus AC4 protein is required for membrane binding and pathogenicity. Molecular Plant Microbe Interact. 2007;20(4):380–91.
Chowda-Reddy RV, Achenjang F, Feltona C, Etarocka MT, Anangfaca MT, Patricia Nugenta P, Fondong VN. Role of a geminivirus AV2 protein putative protein kinase C motif on subcellular localization and pathogenicity. Virus Res. 2008;135:115–24.
Mansoor S, et al. Geminivirus disease complexes: an emerging threat. Trends Plant Sci. 2003;8:128–34.
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The First author is grateful to National Agricultural higher Education Project of Indian Council of Agricultural Research for extending support and providing research grant for the study.
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Sarkar, M., Aggarwal, S., Mukherjee, S.K. et al. Sub-cellular localization of suppressor proteins of tomato leaf curl New Delhi virus. VirusDis. 32, 298–304 (2021). https://doi.org/10.1007/s13337-021-00651-0
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DOI: https://doi.org/10.1007/s13337-021-00651-0