The C4 protein of tomato yellow leaf curl Sardinia virus primes drought tolerance in tomato through morphological adjustments

Abstract

Viruses can interfere with the ability of plants to overcome abiotic stresses, indicating the existence of common molecular networks that regulate stress responses. A begomovirus causing the tomato yellow leaf curl disease was recently shown to enhance heat tolerance in tomato and drought tolerance in tomato and Nicotiana benthamiana and experimental evidence suggested that the virus-encoded protein C4 is the main trigger of drought responses. However, the physiological and molecular events underlying C4-induced drought tolerance need further elucidation. In this study, transgenic tomato plants expressing the tomato yellow leaf curl Sardinia virus (TYLCSV) C4 protein were subjected to severe drought stress, followed by recovery. Morphometric parameters, water potential, gas exchanges, and hormone contents in leaves were measured, in combination with molecular analysis of candidate genes involved in stress response and hormone metabolism. Collected data proved that the expression of TYLCSV C4 positively affected the ability of transgenic plants to tolerate water stress, by delaying the onset of stress-related features, improving the plant water use efficiency and facilitating a rapid post-rehydration recovery. In addition, we demonstrated that specific anatomical and hydraulic traits, rather than biochemical signals, are the keynote of the C4-associated stress resilience. Our results provide novel insights into the biology underpinning drought tolerance in TYLCSV C4-expressing tomato plants, paving the way for further deepening the mechanism through which such proteins tune the plant-virus interaction.

References

[1] Rosenzweig C, Elliott J, Deryng D et al. Assessing agricultural risks of climate change in the 21st century in a global gridded crop model intercomparison. Proc Natl Acad Sci U S A. 2014; 111: 3268-73.

[2] Takahashi F, Kuromori T, Urano K et al. Drought stress responses and resistance in plants: from cellular responses to long-distance intercellular communication. Front Plant Sci. 2020; 11: 556972.

[3] Lovisolo C, Perrone I, Hartung W et al. An abscisic acid-related reduced transpiration promotes gradual embolism repair when grapevines are rehydrated after drought. New Phytol. 2008; 180: 642-51.

[4] Kuromori T, Seo M, Shinozaki K . ABA transport and plant water stress responses. Trends Plant Sci. 2018; 23: 513-22.

[5] Claeys H, Inzé D . The agony of choice: how plants balance growth and survival under water-limiting conditions. Plant Physiol. 2013; 162: 1768-79.

[6] Hirayama T, Shinozaki K . Research on plant abiotic stress responses in the post-genome era: past, present and future. Plant J. 2010; 61: 1041-52.

[7] Zhou R, Kong L, Wu Z et al. Physiological response of tomatoes at drought, heat and their combination followed by recovery. Physiol Plant. 2019; 165: 144-54.

[8] Panno S, Davino S, Caruso AG et al. A review of the most common and economically important diseases that undermine the cultivation of tomato crop in the Mediterranean basin. Agronomy. 2021; 11: 2188.

[9] Chattopadhyay C, Birah A, Jalali BL . Climate change: impact on biotic stresses afflicting crop plants. In: Peshin R, Dhawan A, eds. Natural Resource Management: Ecological Perspectives. Sustainability in Plant and Crop Protection.Springer: Charm, 2019, 133-46.

[10] Rahman A, Sinha KV, Sopory SK et al. Influence of virus-host interactions on plant response to abiotic stress. Plant Cell Rep. 2021; 40: 2225-45.

[11] Prasad A, Sett S, Prasad M . Plant-virus-abiotic stress interactions: a complex interplay. Environ Exp Bot. 2022; 199: 104869.

[12] Yan Z, Wolters AA, Navas-Castillo J et al. The global dimension of tomato yellow leaf curl disease: current status and breeding perspectives. Microorganisms. 2021; 9: 740.

[13] Kheyr-Pour A, Bendahmane M, Matzeit V et al. Tomato yellow leaf curl virus from Sardinia is a whitefly-transmitted monopartite geminivirus. Nucl Acids Res. 1991; 19: 6763-9.

[14] Medina-Puche L, Orìlio AF, Zerbini FM et al. Small but mighty: functional landscape of the versatile geminivirus encoded C4 protein. PLoS Pathog. 2021; 17: e1009915.

[15] De Jupin I, Kouchkovsky F, Jouanneau F et al. Movement of tomato yellow leaf curl geminivirus (TYLCV): involvement of the protein encoded by ORF C4. Virology. 1994; 204: 82-90.

[16] Rojas MR, Jiang H, Salati R et al. Functional analysis of proteins involved in movement of the monopartite begomovirus, tomato yellow leaf curl virus. Virology. 2001; 291: 110-25.

[17] Luna AP, Morilla G, Voinnet O et al. Functional analysis of gene-silencing suppressors from tomato yellow leaf curl disease viruses. Mol Plant-Microbe Interact. 2012; 25: 1294-306.

[18] Rosas-Diaz T, Zhang D, Fan P et al. A virus-targeted plant receptor-like kinase promotes cell-to-cell spread of RNAi. Proc Natl Acad Sci U S A. 2018; 115: 1388-93.

[19] Rigden JE, Krake LR, Rezaian MA et al. ORF C4 of tomato leaf curl geminivirus is a determinant of symptom severity. Virology. 1994; 204: 847-50.

[20] Mei Y, Ma Z, Wang Y et al. Geminivirus C4 antagonizes the HIR-mediated hypersensitive response by inhibiting the HIR1 self-interaction and promoting degradation of the protein. New Phytol. 2020; 225: 1311-26.

[21] Jing C, Li P, Zhang J et al. The Malvastrum yellow vein virus C4 protein promotes disease symptom development and enhances virus accumulation in plants. Front Microbiol. 2019; 10: 2425.

[22] Aguilar E, Cutrona C, del Toro FJ et al. Virulence determines beneficial trade-offs in the response of virus-infected plants to drought via induction of salicylic acid. Plant Cell Environ. 2017; 40: 2909-30.

[23] Carr JP . Exploring how viruses enhance plants’ resilience to drought and the limits to this form of viral payback. Plant Cell Environ. 2017; 40: 2906-8.

[24] Xu P, Chen F, Mannas JP et al. Virus infection improves drought tolerance. New Phytol. 2008; 180: 911-21.

[25] Bostock RM, Pye MF, Roubtsova TV . Predisposition in plant disease: exploiting the nexus in abiotic and biotic stress perception and response. Annu Rev Phytopathol. 2014; 52: 517-49.

[26] Westwood JH, Mccann L, Naish M et al. A viral RNA silencing suppressor interferes with abscisic acid-mediated signalling and induces drought tolerance in Arabidopsis thaliana. Mol Plant Pathol. 2013; 14: 158-70.

[27] Pantaleo V, Vitali M, Boccacci P et al. Novel functional microRNAs from virus-free and infected Vitis vinifera plants under water stress. Sci Rep. 2016; 6: 20167.

[28] Ascencio-Ibáñez JT, Sozzani R, Lee TJ et al. Global analysis of Arabidopsis gene expression uncovers a complex array of changes impacting pathogen response and cell cycle during geminivirus infection. Plant Physiol. 2008; 148: 436-54.

[29] Hanley-Bowdoin L, Bejarano ER, Robertson D et al. Geminiviruses: masters at redirecting and reprogramming plant processes. Nat Rev Microbiol. 2013; 11: 777-88.

[30] Chen T, Lv Y, Zhao T et al. Comparative transcriptome profiling of a resistant vs. susceptible tomato (Solanum lycopersicum) cultivar in response to infection by tomato yellow leaf curl virus. PLoS One. 2013; 8: e80816.

[31] Miozzi L, Napoli C, Sardo L et al. Transcriptomics of the interaction between the monopartite phloem-limited geminivirus tomato yellow leaf curl Sardinia virus and Solanum lycopersicum highlights a role for plant hormones, autophagy and plant immune system fine tuning during infection. PLoS One. 2014; 9: e89951.

[32] Anfoka G, Moshe A, Fridman L et al. Tomato yellow leaf curl virus infection mitigates the heat stress response of plants grown at high temperatures. Sci Rep. 2016; 6: 19715.

[33] Corrales-Gutierrez M, Medina-Puche L, Yu Y et al. The C4 protein from the geminivirus tomato yellow leaf curl virus confers drought tolerance in Arabidopsis through an ABA-independent mechanism. Plant Biotech J. 2019; 18: 1121-3.

[34] Shteinberg M, Mishra R, Anfoka G et al. Tomato yellow leaf curl virus (TYLCV) promotes plant tolerance to drought. Cell. 2021; 10: 2875.

[35] Mishra R, Shteinberg M, Shkolnik D et al. Interplay between abiotic (drought) and biotic (virus) stresses in tomato plants. Mol Plant Pathol. 2022; 23: 475-88.

[36] Chitarra W, Pagliarani C, Maserti B et al. Insights on the impact of arbuscular mycorrhizal symbiosis on tomato tolerance to water stress. Plant Physiol. 2016; 171: 1009-23.

[37] 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. Virus Disease. 2017; 28: 390-400.

[38] Raghavendra AS, Gonugunta VK, Christmann A et al. ABA perception and signalling. Trends Plant Sci. 2010; 15: 395-401.

[39] Visentin I, Pagliarani C, Deva E et al. A novel strigolactone-miR156 module controls stomatal behaviour during drought recovery. Plant Cell Environ. 2020; 43: 1613-24.

[40] Sacco A, Greco B, Di Matteo A et al. Evaluation of tomato genetic resources for response to water deficit. Am J Plant Sci. 2013; 04: 131-45.

[41] Yoshiba Y, Kiyosue T, Nakashima K et al. Regulation of levels of proline as an osmolyte in plants under water stress. Plant Cell Physiol. 1997; 38: 1095-102.

[42] Hu C, Delauney AJ, Verma D . A bifunctional enzyme (delta 1-pyrroline-5-carboxylate synthetase) catalyzes the first two steps in proline biosynthesis in plants. Proc Natl Acad Sci U S A. 1992; 89: 9354-8.

[43] Kiyosue T, Yoshiba Y, Yamaguchi-Shinozaki K et al. A nuclear gene encoding mitochondrial proline dehydrogenase, an enzyme involved in proline metabolism, is upregulated by proline but downregulated by dehydration in Arabidopsis. Plant Cell. 1996; 8: 1323-35.

[44] Bertolino LT, Caine RS, Gray JE . Impact of Stomatal density and morphology on water-use efficiency in a changing world. Front Plant Sci. 2019; 10: 225.

[45] Fan P, Aguilar E, Bradai M et al. The receptor-like kinases BAM1 and BAM2 are required for root xylem patterning. Proc Natl Acad Sci U S A. 2021; 118: e2022547118.

[46] López R, Cano FJ, Martin-StPaul NK et al. Coordination of stem and leaf traits define different strategies to regulate water loss and tolerance ranges to aridity. New Phytol. 2021; 230: 497-509.

[47] Lovisolo C, Schubert A . Effects of water stress on vessel size and xylem hydraulic conductivity in Vitis vinifera L. J Exp Bot. 1998; 49: 693-700.

[48] Zouine M, Fu Y, Chateigner-Boutin AL et al. Characterization of the tomato ARF gene family uncovers a multi-levels post-transcriptional regulation including alternative splicing. PLoS One. 2014; 9: e84203.

[49] Liu S, Zhang Y, Feng Q et al. Tomato AUXIN RESPONSE FACTOR 5 regulates fruit set and development via the mediation of auxin and gibberellin signaling. Sci Rep. 2018; 8: 2971.

[50] Ghosh D, Chakraborty S . Molecular interplay between phytohormones and geminiviruses: a saga of a never-ending arms race. J Exp Bot. 2021; 72: 2903-17.

[51] Wu M, Ding X, Fu X et al. Transcriptional reprogramming caused by the geminivirus tomato yellow leaf curl virus in local or systemic infections in Nicotiana benthamiana. BMC Genomics. 2019; 20: 542.

[52] Vinutha T, Vanchinathan S, Bansal N et al. Tomato auxin biosynthesis/signaling is reprogrammed by the geminivirus to enhance its pathogenicity. Planta. 2020; 252: 51.

[53] Bouzroud S, Gouiaa S, Hu N et al. Auxin response factors (ARFs) are potential mediators of auxin action in tomato response to biotic and abiotic stress (Solanum lycopersicum). PLoS One. 2018; 13: e0193517.

[54] Fraile A, García-Arenal F . The coevolution of plants and viruses: resistance and pathogenicity. Natural and Engineered Resistance to Plant Viruses, Part II. 2010; 76: 1-32.

[55] Dastogeer K. Influence of fungal endophytes on plant physiology is more pronounced under stress than well-watered conditions: a meta-analysis. Planta. 2018; 248: 1403-16.

[56] Conrath U, Pieterse CMJ, Mauch-Mani B . Priming in plant pathogen interactions. Trends Plant Sci. 2002; 7: 210-6.

[57] Brunetti A, Tavazza M, Noris E et al. High expression of truncated viral rep protein confers resistance to tomato yellow leaf curl virus in transgenic tomato plants. Mol Plant-Microbe Interact. 1997; 10: 571-9.

[58] Noris E, Accotto GP, Tavazza R et al. Resistance to tomato yellow leaf curl geminivirus in Nicotiana benthamiana plants transformed with a truncated viral C1 gene. Virology. 1996; 224: 130-8.

[59] Manacorda CA, Gudesblat G, Sutka M et al. TuMV triggers stomatal closure but reduces drought tolerance in Arabidopsis. Plant Cell Environ. 2021; 44: 1399-416.

[60] Hopper DW, Ghan R, Cramer GR . A rapid dehydration leaf assay reveals stomatal response differences in grapevine genotypes. Hortic Res. 2014; 1: 2.

[61] Kardiman R, Ræbild R . Relationship between stomatal density, size and speed of opening in Sumatran rainforest species. Tree Physiol. 2018; 38: 696-705.

[62] Pagliarani C, Gambino G, Ferrandino A et al. Molecular memory of Flavescence dorée phytoplasma in recovering grapevines. Hortic Res. 2020; 7: 126.

[63] Mannino G, Nerva L, Gritli T et al. Effects of different microbial inocula on tomato tolerance to water deficit. Agronomy. 2020; 10: 170.

[64] Callister AN, Arndt SK, Adams MA . Comparison of four methods for measuring osmotic potential of tree leaves. Physiol Plant. 2006; 127: 383-92.

[65] Livak KJ, Schmittgen TD . Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods. 2001; 25: 402-8.