Photosynthetica X:X | DOI: 10.32615/ps.2026.002

Impact of tetraploidization on morphophysiological leaf traits in the drought tolerant 'de Ramellet' tomato landrace

P. CERDÁ-BENNASSER1, M. FULLANA-PERICÀS1, P. AGUILÓ-NICOLAU1, J. PELLICER2, 3, J. GALMÉS1, M.À. CONESA1
1 Agro-Environmental and Water Economics Research Institute (INAGEA) - Research Group on Plant Biology Under Mediterranean Conditions (PlantMed) - Department of Biology, University of the Balearic Islands (UIB), Balearic Complex for Research, Technological Development and Innovation, Blaise Pascal 6, 07121 Palma (Illes Balears), Spain
2 Botanical Institute of Barcelona (IBB), CSIC-CMCNB, Passeig del Migdia s.n., 08038 Barcelona, Spain
3 Royal Botanic Gardens, Kew, Richmond TW9 3AE, United Kingdom

Tetraploidization was induced in the drought-tolerant tomato landrace 'de Ramellet' to evaluate its physiological and anatomical response under well-watered (WW) and water-deficient (WD) conditions. Under WW, tetraploid plants exhibited approximately 40% lower stomatal density and approximately 80% larger stomata than diploids. Net photosynthetic rate (PN), intrinsic water-use efficiency, and intercellular CO2 concentration remained unchanged between diploids and tetraploids. Under WD, both genotypes reduced PN and stomatal conductance by similar proportions; however, only diploids decreased leaf area and adjusted stomatal density and size, whereas the tetraploid maintained stomatal traits similarly to those in WW conditions. However, both genotypes maintained similar photosynthetic capacity under WD despite different stomatal display and total pore area, which suggests the involvement of morphophysiological mechanisms beyond stomatal traits, such as root traits and hydraulic regulation.

Additional key words: drought stress; gas exchange traits; Solanum lycopersicum; stomatal traits; tetraploidization.

Received: October 21, 2025; Revised: December 17, 2025; Accepted: January 7, 2026; Prepublished online: January 20, 2026 

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    References

    1. Aryavand A., Ehdaie B., Tran B. et al.: Stomatal frequency and size differentiate ploidy levels in Aegilops neglecta. - Genet. Resour. Crop Evol. 50: 175-182, 2003. Go to original source...
    2. Assmann S., Zeiger E.: Guard cell bioenergetics. - In: Zeiger E., Farquhar G.D., Cowan I.R. (ed.): Stomatal Function. Pp. 163-193. Stanford University Press, Palo Alto 1987.
    3. Beaulieu J.M., Leitch I.J., Patel S. et al.: Genome size is a strong predictor of cell size and stomatal density in angiosperms. - New Phytol. 179: 975-986, 2008. Go to original source...
    4. Bertolino L.T., Caine R.S., Gray J.E.: Impact of stomatal density and morphology on water-use efficiency in a changing world. -Front. Plant Sci. 10: 225, 2019. Go to original source...
    5. Caine R., Yin X., Sloan J. et al.: Rice with reduced stomatal density conserves water and has improved drought tolerance under future climate conditions. - New Phytol. 221: 371-384, 2019. Go to original source...
    6. Capomaccio S., Veronesi F., Rosellini D.: Polyploidization and gene expression in Medicago sativa. - In: Huyghe C. (ed.): Sustainable Use of Genetic Diversity in Forage and Turf Breeding. Pp. 397-401. Springer, Dordrecht 2010. Go to original source...
    7. Chaves M.M., Costa J.M., Zarrouk O. et al.: Controlling stomatal aperture in semi-arid regions - The dilemma of saving water or being cool? - Plant Sci. 251: 54-64, 2016. Go to original source...
    8. Chen Z.J., Sreedasyam A., Ando A. et al.: Genomic diversifications of five Gossypium alloplyploid species and their impact on cotton improvement. - Nat. Genet. 52: 525-533, 2020. Go to original source...
    9. Conesa M.À., Fullana-Pericàs M., Granell A., Galmés J.: Mediterranean long shelf-life landraces: an untapped genetic resource for tomato improvement. - Front. Plant Sci 10: 1651, 2020. Go to original source...
    10. Dow G.J., Bergmann D.C., Berry J.A.: An integrated model of stomatal development and leaf physiology. - New Phytol. 201: 1218-1226, 2014. Go to original source...
    11. Drake P.L., de Boer H.J., Schymanski S.J., Veneklaas E.J.: Two sides to every leaf: water and CO2 transport in hypostomatous and amphistomatous leaves. - New Phytol. 222: 1179-1187, 2019. Go to original source...
    12. Drake P.L., Froend R.H., Franks P.J.: Smaller, faster stomata: scaling of stomatal size, rate of response, and stomatal conductance. - J. Exp. Bot. 64: 495-505, 2013. Go to original source...
    13. Driesen E., De Proft M., Saeys W.: Drought stress triggers alterations of adaxial and abaxial stomatal development in basil leaves increasing water-use efficiency. - Hortic. Res. 10: uhad075, 2023. Go to original source...
    14. Edger P.P., Poorten T.J., VanBuren R. et al.: Origin and evolution of the octoploid strawberry genome. - Nat. Genet. 51: 541-547, 2019. Go to original source...
    15. Feldman M., Lupton F.G.H., Miller T.E.: Wheats. - In: Smartt J., Simmonds N. (ed.): Evolution of Crop Plants. Pp. 184-192. Wiley-Blackwell, Harlow 1995.
    16. Firon N., Shaked R., Peet M.M. et al.: Pollen grains of heat tolerant tomato cultivars retain higher carbohydrate concentration under heat stress conditions. - Sci. Hortic.-Amsterdam 109: 212-217, 2006. Go to original source...
    17. Flexas J., Ortuño M.F., Ribas-Carbo M. et al.: Mesophyll conductance to CO2 in Arabidopsis thaliana. - New Phytol. 175: 501-511, 2007. Go to original source...
    18. Franks P.J., Beerling D.J.: Maximum leaf conductance driven by CO2 effects on stomatal size and density over geologic time. - PNAS 106: 10343-10347, 2009. Go to original source...
    19. Franks P.J., Farquhar G.D.: The mechanical diversity of stomata and its significance in gas-exchange control. - Plant Physiol. 143: 78-87, 2007. Go to original source...
    20. Fullana-Pericàs M., Conesa M.À., Douthe C. et al.: Tomato landraces as a source to minimize yield losses and improve fruit quality under water deficit conditions. - Agr. Water Manage. 223: 105722, 2019. Go to original source...
    21. Fullana-Pericàs M., Conesa M.À., Gago J. et al.: High-throughput phenotyping of a large tomato collection under water deficit: Combining UAVs' remote sensing with conventional leaf-level physiologic and agronomic measurements. - Agr. Water Manage. 260: 107283, 2022. Go to original source...
    22. Fullana-Pericàs M., Conesa M.À., Soler S. et al.: Variations of leaf morphology, photosynthetic traits and water-use efficiency in Western-Mediterranean tomato landraces. - Photosynthetica 55: 121-133, 2017. Go to original source...
    23. Galmés J., Conesa M.À., Ochogavía J.M. et al.: Physiological and morphological adaptations in relation to water use efficiency in Mediterranean accessions of Solanum lycopersicum. - Plant Cell Environ. 34: 245-260, 2011. Go to original source...
    24. Galmés J., Ochogavía J.M., Gago J. et al.: Leaf responses to drought stress in Mediterranean accessions of Solanum lycopersicum: anatomical adaptations in relation to gas exchange parameters. - Plant Cell Environ. 36: 920-935, 2013. Go to original source...
    25. Haworth M., Marino G., Materassi A. et al.: The functional significance of the stomatal size to density relationship: Interaction with atmospheric [CO2] and role in plant physiological behaviour. - Sci. Total Environ. 863: 160908, 2023. Go to original source...
    26. Haworth M., Scutt C.P., Douthe C. et al.: Allocation of the epidermis to stomata relates to stomatal physiological control: Stomatal factors involved in the evolutionary diversification of the angiosperms and development of amphistomaty. - Environ. Exp. Bot. 151: 55-63, 2018. Go to original source...
    27. Hetherington A.M., Woodward F.I.: The role of stomata in sensing and driving environmental change. - Nature 424: 901-908, 2003. Go to original source...
    28. Lawson T., Blatt M.R.: Stomatal size, speed, and responsiveness impact on photosynthesis and water use efficiency. - Plant Physiol. 164: 1556-1570, 2014. Go to original source...
    29. Lawson T., Vialet-Chabrand S.: Speedy stomata, photosynthesis and plant water use efficiency. - New Phytol. 221: 93-98, 2019. Go to original source...
    30. Levy A.A., Feldman M.: Evolution and origin of bread wheat. - Plant Cell 34: 2549-2567, 2022. Go to original source...
    31. Lindstrom E.W., Humphrey L.M.: Comparative cyto-genetic studies of tetraploid tomatoes from different origins. - Genetics 18: 193-209, 1933. Go to original source...
    32. Loureiro J., Rodriguez E., Dole¾el J., Santos C.: Two new nuclear isolation buffers for plant DNA flow cytometry: a test with 37 species. - Ann. Bot.-London 100: 875-888, 2007. Go to original source...
    33. Lu Y., Duursma R.A., Farrior C.E. et al.: Optimal stomatal drought response shaped by competition for water and hydraulic risk can explain plant trait covariation. - New Phytol. 225: 1206-1217, 2020. Go to original source...
    34. Lüttge U.: Ability of crassulacean acid metabolism plants to overcome interacting stresses in tropical environments. - AoB Plants 2010: plq005, 2010. Go to original source...
    35. Massman W.J.: A review of the molecular diffusivities of H2O, CO2, CH4, CO, O3, SO2, NH3, N2O, NO, and NO2 in air, O2 and N2 near STP. - Atmos. Environ. 32: 1111-1127, 1998. Go to original source...
    36. Masterson J.: Stomatal size in fossil plants: evidence for polyploidy in majority of angiosperms. - Science 264: 421-424, 1994. Go to original source...
    37. McAusland L., Vialet-Chabrand S., Davey P. et al.: Effects of kinetics of light-induced stomatal responses on photosynthesis and water-use efficiency. - New Phytol. 211: 1209-1220, 2016. Go to original source...
    38. McElwain J.C., Yiotis C., Lawson T.: Using modern plant trait relationships between observed and theoretical maximum stomatal conductance and vein density to examine patterns of plant macroevolution. - New Phytol. 209: 94-103, 2016. Go to original source...
    39. Mott K.A.: Leaf hydraulic conductivity and stomatal responses to humidity in amphistomatous leaves. - Plant Cell Environ. 30: 1444-1449, 2007. Go to original source...
    40. Muir C.D.: Making pore choices: repeated regime shifts in stomatal ratio. - Proc. Biol. Sci. 282: 20151498, 2015. Go to original source...
    41. Muir C.D.: Light and growth form interact to shape stomatal ratio among British angiosperms. - New Phytol. 218: 242-252, 2018. Go to original source...
    42. Muir C.D., Conesa M.À., Roldán E.J. et al.: Weak coordination between leaf structure and function among closely related tomato species. - New Phytol. 213: 1642-1653, 2017. Go to original source...
    43. Nilsson E.: Some experiments with tetraploid tomatoes. - Hereditas 36: 181-204, 1950. Go to original source...
    44. OTPTI (One Thousand Plant Transcriptomes Initiative): One thousand plant transcriptomes and the phylogenomics of green plants. - Nature 574: 679-685, 2019. Go to original source...
    45. Pellicer J., Powell R.F., Leitch I.J.: The application of flow cytometry for estimating genome size, ploidy level endopolyploidy, and reproductive modes in plants. - In: Besse P. (ed.): Molecular Plant Taxonomy. Methods in Molecular Biology. Vol. 2222. Pp. 325-361. Humana, New York 2021. Go to original source...
    46. Peppe D.J., Royer D.L., Cariglino B. et al.: Sensitivity of leaf size and shape to climate: global patterns and paleoclimatic applications. - New Phytol. 190: 724-739, 2011. Go to original source...
    47. Pitaloka M.K., Caine R.S., Hepworth C. et al.: Induced genetic variations in stomatal density and size of rice strongly affects water use efficiency and responses to drought stresses. - Front. Plant Sci. 13: 801706, 2022. Go to original source...
    48. Praça M.M., Carvalho C.R., Clarindo W.R.: A practical and reliable procedure for in vitro induction of tetraploid tomato. -Sci. Hortic.-Amsterdam 122: 501-505, 2009. Go to original source...
    49. R Core Team: R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna 2024. Available at: https://www.R-project.org.
    50. Rice A., ©marda P., Novosolov M. et al.: The global biogeography of polyploid plants. - Nat. Ecol. Evol. 3: 265-273, 2019. Go to original source...
    51. Richardson F., Brodribb T.J., Jordan G.J.: Amphistomatic leaf surfaces independently regulate gas exchange in response to variations in evaporative demand. - Tree Physiol. 37: 869-878, 2017. Go to original source...
    52. Russo S.E., Cannon W.L., Elowsky C. et al.: Variation in leaf stomatal traits of 28 tree species in relation to gas exchange along an edaphic gradient in a Bornean rain forest. - Am. J. Bot. 97: 1109-1120, 2010. Go to original source...
    53. Sack L., Buckley T.N.: The developmental basis of stomatal density and flux. - Plant Physiol. 171: 2358-2363, 2016. Go to original source...
    54. Sato S., Peet M.M., Thomas J.F.: Determining critical pre- and post-anthesis periods and physiological processes in Lycopersicon esculentum Mill. exposed to moderately elevated temperatures. - J. Exp. Bot. 53: 1187-1195, 2002. Go to original source...
    55. Sattler M.C., Carvalho C.R., Clarindo W.R.: The polyploidy and its key role in plant breeding. - Planta 243: 281-296, 2016. Go to original source...
    56. Scalabrin S., Toniutti L., Di Gaspero G. et al.: A single polyploidization event at the origin of the tetraploid genome of Coffea arabica is responsible for the extremely low genetic variation in wild and cultivated germplasm. - Sci. Rep.-UK 10: 4642, 2020. Go to original source...
    57. Song L., Ding R., Du T. et al.: Stomatal conductance parameters of tomatoes are regulated by reducing osmotic potential and pre-dawn leaf water potential via increasing ABA under salt stress. - Environ. Exp. Bot. 206: 105176, 2023. Go to original source...
    58. Srivastava A., Lu Z., Zeiger E.: Modification of guard cell properties in advanced lines of Pima cotton bred for higher yields and heat resistance. - Plant Sci. 108: 125-131, 1995. Go to original source...
    59. Suda J., Krahulcová A., Trávníèek P., Krahulec F.: Ploidy level versus DNA ploidy level: an appeal for consistent terminology. - Taxon 55: 447-450, 2006. Go to original source...
    60. Tate J.A., Soltis D.E., Soltis P.S.: Polyploidy in plants. - In: Gregory T.R. (ed.): The Evolution of the Genome. Pp. 371-402. Academic Press, San Diego 2004. Go to original source...
    61. Tomaszewska P., Schwarzacher T., Heslop-Harrison J.S.: Oat chromosome and genome evolution defined by widespread terminal intergenomic translocations in polyploids. - Front. Plant Sci. 13: 1026364, 2022. Go to original source...
    62. Trojak-Goluch A., Kawka-Lipiñska M., Wielgusz K., Praczyk M.: Polyploidy in industrial crops: applications and perspectives in plant breeding. - Agronomy 11: 2574, 2021. Go to original source...
    63. Van de Peer Y., Ashman T.-L., Soltis P.S., Soltis D.E.: Polyploidy: an evolutionary and ecological force in stressful times. - Plant Cell 33: 11-26, 2021. Go to original source...
    64. Van Laere K., França S.C., Vansteenkiste H. et al.: Influence of ploidy level on morphology, growth and drought susceptibility in Spathiphyllum wallisii. - Acta Physiol. Plant. 33: 1149-1156, 2011. Go to original source...
    65. Wang Y., Noguchi K., Terashima I.: Photosynthesis-dependent and -independent responses of stomata to blue, red and green monochromatic light: differences between the normally oriented and inverted leaves of sunflower. - Plant Cell Physiol. 52: 479-489, 2011. Go to original source...
    66. Wilson M.J., Fradera-Soler M., Summers R. et al.: Ploidy influences wheat mesophyll cell geometry, packing and leaf function. - Plant Direct 5: e00314, 2021. Go to original source...
    67. Xiong D., Flexas J.: From one side to two sides: the effects of stomatal distribution on photosynthesis. - New Phytol. 228: 1754-1766, 2020. Go to original source...
    68. Zhang K., Wang X., Cheng F.: Plant polyploidy: origin, evolution, and its influence on crop domestication. - Hortic. Plant J. 5: 231-239, 2019. Go to original source...
    69. Zhang L., Ji H., Jiang M. et al.: Recent insights into molecular breeding for high yield sweet potato cultivars. - Biosci. Methods 16: 23-32, 2025. Go to original source...

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