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Abstract
Degenia velebitica (Degen) Hayek, the most important representative of the Croatian endemic flora, is a heliophyte and xerophyte highly specialized for survival in extreme living conditions characterised by intense solar radiation. In this work, the composition and in situ localization of flavonoids in the epidermal trichomes and leaf tissues of D. velebitica were investigated. Flavonoids were localized in the lumens of stellate trichomes, vacuoles and cell walls of epidermal cells, but only in peripheral parts of spongy and palisade parenchyma cells, on both the abaxial and adaxial sides of the leaf. Quercetin, kaempferol and isorhamnetin were the main flavonoid aglycones in extracts of trichomes and leaves. The results show the important physiological role of epidermal trichomes and flavonoids in protecting D. velebitica against insolation.
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Copyright (c) 2025 Vanja Stamenković, Mirta Tkalec

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References
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- Agati, G., Stefano, G., Biricolti, S., Tattini, M., 2009: Mesophyll distribution of ‘antioxidant’flavonoid glycosides in Ligustrum vulgare leaves under contrasting sunlight irradiance. Annals of Botany 104(5), 853–861. https://doi.org/10.1093/aob/mcp177
- Agati, G., Tattini, M., 2010: Multiple functional roles of flavonoids in photoprotection. New Phytologist 186(4), 786–793. https://doi.org/10.1111/j.1469-8137.2010.03269.x
- Ančev, M., Goranova, V., 2006: Trichome morphology of eleven genera of the tribe Alysseae (Brassicaceae) occurring in Bulgaria. Willdenowia 36(1), 193–204.
- Bucar, F., Plazibat, M., Knauder, E., Maleš, Ž., 2005: LC/PDA/MS/MS analysis of flavonoids in some taxa of Alyssum and Aurinia (Brassicaceae) from Croatia. In: Zorc, B. (ed.), Book of Abstracts of the Third Croatian Congress on Pharmacy with International Participation, 188–188. Faculty of Pharmacy and Biochemistry, Zagreb.
- Damjanović, O., Stevanović, B., 1993: Morpho-anatomical adaptations of endemic species Fibigia triquetra (DC.) Boiss. (Brassicaceae). Bulletin de l’Institut et du Jardin Botaniques de l’Université de Beograd 24-25, 9–19.
- Degen, A., 1909: Bemerkungen über einige orientalische Pflanzarten. LIII. Über die Entdeckung eines Vertreters der Gattung Lesquerella im Velebitgebirge. Magyar Botanikai Lapok 8, 3–24
- Domac, R., 1993: Degenia Hayek. In: Tutin, T. G., Burges, N. A., Chater A. O., Edmondson J. R., Heywood, V., Moore, D. M., Valentine, D. H., Walters, S. M., Webb, D. A. (eds.), Flora Europaea, 359, vol. 1. 2nd ed. Cambridge University Press, Cambridge.
- Dunkić, V., Bezić, N., Mileta, T., 2001: Xeromorphism of trichomes in Lamiaceae species. Acta Botanica Croatica 60(2), 277–283.
- Fahn, A., Cutler, F. D., 1992: Xerophytes. Encyclopedia of Plant Anatomy, vol. 3. Gebrüder Bornträger, Berlin, Stuttgart.
- Hayek, A., 1910: Die systematische Stellung von Lesquerella velebitica Degen. Oesterreichische botanische Zeitschrift 60, 89–93
- Hopewell, T., Selvi, F., Ensikat, H. J., Weigend, M., 2021: Trichome biomineralization and soil chemistry in Brassicaceae from Mediterranean ultramafic and calcareous soils. Plants 10(2), 377. https://doi.org/10.3390/plants10020377
- Horvat, I., 1930: Vegetacijske studije o hrvatskim planinama. Zadruge na planinskim goletima [Vegetation studies on Croa-tian mountains. Communities on mountain bare areas]. Rad Jugoslavenske Akademije Znanosti 238, 1–96.
- Horvat, I, 1931: Vegetacijske studije o hrvatskim planinama. II. Zadruge na planinskim stijenama i točilima [Vegetation studies on Croatian mountains. II. Communities on mountain rocks and screes]. Rad Jugoslavenske Akademije Znanosti 241, 147–206.
- Hutzler, P., Fischbach, R., Heller, W., Jungblut, T., Reuber, S., Schmitz, R., Veit, M., Weissenböck, G., Schnitzler, J., 1998: Tissue localization of phenolic compounds in plants by confocal laser scanning microscopy. Journal of Experimental Botany 49(323), 953–965. https://doi.org/10.1093/jxb/49.323.953
- Karabourniotis, G., Liakopoulos, G., Nikolopoulos, D., Bresta, P., 2020: Protective and defensive roles of non-glandular trichomes against multiple stresses: structure–function coordination. Journal of Forestry Research 31(1), 1-12. https://doi.org/10.1007/s11676-019-01034-4
- Laoué, J., Fernandez, C., Ormeño, E., 2022: Plant flavonoids in mediterranean species: A focus on flavonols as protective metabolites under climate stress. Plants 11(2), 172. https://doi.org/10.3390/plants11020172
- Liakopoulos, G., Stavrianakou, S., Karabourniotis, G., 2006: Trichome layers versus dehaired lamina of Olea europaea leaves: differences in flavonoid distribution, UV-absorbing capacity, and wax yield. Environmental and Experimental Botany 55(3), 294–304. https://doi.org/10.1016/j.envexpbot.2004.11.008
- Liakoura, V., Stefanou, M., Manetas, Y., Cholevas, C., Karabourniotis, G., 1997: Trichome density and its UV-B protective potential are affected by shading and leaf position on the canopy. Environmental and Experimental Botany 38(3), 223–229. http://dx.doi.org/10.1016/S0098-8472(97)00005-1
- Liber, Z., Surina, B., Nikolić, T., Škrtić, D., Šatović, Z., 2020: Spatial distribution, niche ecology and conservation genetics of Degenia velebitica (Brassicaceae), a narrow endemic species of the north-western Dinaric Alps. Plant Systematics and Evolution 306(4), 64. https://doi.org/10.1007/s00606-020-01695-3
- Manetas, Y., 2003: The importance of being hairy: the adverse effects of hair removal on stem photosynthesis of Ver-bascum speciosum are due to solar UV-B radiation. New Phytologist 158, 503–508.
- Mastelić, J., Blažević, I., Kosalec, I., 2010: Chemical composition and antimicrobial activity of volatiles from Degenia vele-bitica, a European stenoendemic plant of the Brassicaceae family. Chemistry & Biodiversity, 7(11), 2755–2765. https://doi.org/10.1002/cbdv.201000053
- Naumovski, D., 2005: Germination ecology of seeds of endemic species Degenia velebitica (Degen) Hayek (Brassicaceae). Acta Botanica Croatica 64(2), 323–330.
- Nikolić, T., Milović, M., Bogdanović, S., Jasprica, N., 2015: Endemi u hrvatskoj flori [Endemics in Croatian flora]. Alfa d.d., Zagreb.
- Nikolić, T. (ed.), 2015-2025: Flora Croatica Database. Retrieved April 5, 2025 from http://hirc.botanic.hr/fcd. Pri-rodoslovno-matematički fakultet, Sveučilište u Zagrebu.
- Official Gazette (Narodne Novine), 2013: Pravilnik o strogo zaštićenim vrstama [Ordinance of strictly protected species]. 144. Croatian Government, Zagreb.
- Rešetnik, I., Satovic, Z., Schneeweiss, G. M., Liber, Z., 2013: Phylogenetic relationships in Brassicaceae tribe Alysseae in-ferred from nuclear ribosomal and chloroplast DNA sequence data. Molecular Phylogenetics and Evolution 69(3), 772–786. https://doi.org/10.1016/j.ympev.2013.06.026
- Shen, N., Wang, T., Gan, Q., Liu, S., Wang, L., Jin, B., 2022: Plant flavonoids: Classification, distribution, biosynthesis, and antioxidant activity. Food Chemistry 383, 132531. http://dx.doi.org/10.1016/j.foodchem.2022.132531
- Singh, P., Arif, Y., Bajguz, A., Hayat, S., 2021: The role of quercetin in plants. Plant Physiology and Biochemistry, 166, 10–19. http://dx.doi.org/10.1016/j.plaphy.2021.05.023
- Skaltsa, H., Verykokidou, E., Harvala, C., Karabourniotis, G., Manetasi, Y., 1994: UV-B protective potential and flavonoid content of leaf hairs of Quercus ilex. Phytochemistry 37(4), 987–990. http://dx.doi.org/10.1016/S0031-9422(00)89514-X
- Stamenković, V., Kovačić, S., Juretić, B., Mihelj, D., 2010: Ex-situ protection program for Croatian endemic plant taxa in the Botanical Garden of the Department of Biology, Faculty of Science. In: Jasprica, N., Pandža, M., Milović, M. (eds.), Abstract Book of the Third Croatian Botanical Congress, 180–181. Croatian Botanical Society, Murter, Zagreb.
- Stamenković, V., Likić, S., Tkalec, M., 2015: Defence response of Degenia velebitica (Brassicaceae) to solar UV radiation. In: Bogdanović, S., Jogan, N. (eds.), Book of Abstracts of the Sixth Balkan Botanical Congress, 109–110. Natural History Museum Rijeka, Croatian Botanical Society, Botanical Society of Slovenia, University of Rijeka, Rijeka.
- Stevanović, B., Vujnović, K., 1990: Morpho‐anatomical adaptations of the endemic species Degenia velebitica (Deg.) Hay. Feddes Repertorium 101(7‐8), 385–389. http://dx.doi.org/10.1002/fedr.19901010710
- Sylvester, A. W., Cande, W. Z., Freeling, M., 1991: Developmental morphology of maize leaves using UV light induced autofluorescence of cell wall compounds. Maize Genetics Cooperation Newsletter 65, 34–35.
- Tattini, M., Gravano, E., Pinelli, P., Mulinacci, N., Romani, A., 2000: Flavonoids accumulate in leaves and glandular tri-chomes of Phillyrea latifolia exposed to excess solar radiation. New Phytologist 148(1), 69–77. http://dx.doi.org/10.1046/j.1469-8137.2000.00743.x
- Tattini, M., Matteini, P., Saracini, E., Traversi, M. L., Giordano, C., Agati, G., 2007: Morphology and biochemistry of non-glandular trichomes in Cistus salvifolius L. leaves growing in extreme habitats of the Mediterranean basin. Plant Biology 9(03), 411–419. http://dx.doi.org/10.1055/s-2006-924662
- Tsiftsoglou, O. S., Lazari, D. M., Stefanakis, M. K., Kokkalou, E. L., 2019: Flavonoids and phenolic acids from the aerial parts of Alyssum alyssoides L. (Brassicaceae). Biochemical Systematics and Ecology 83, 51–53. http://dx.doi.org/10.1016/j.bse.2018.12.004
- Watkins, J. M., Chapman, J. M., Muday, G. K., 2017: Abscisic acid-induced reactive oxygen species are modulated by fla-vonols to control stomata aperture. Plant Physiology 175(4), 1807–1825. http://dx.doi.org/10.1104/pp.17.01010
- Watts, S., Kariyat, R., 2021: Morphological characterization of trichomes shows enormous variation in shape, density and dimensions across the leaves of 14 Solanum species. AoB Plants 13(6), plab071. https://doi.org/10.1093/aobpla/plab071
References
Agati, G., Brunetti, C., Fini, A., Gori, A., Guidi, L., Landi, M., Sebastiani, F., Tattini, M., 2020: Are flavonoids effective anti-oxidants in plants? Twenty years of our investigation. Antioxidants 9(11), 1098. https://doi.org/10.3390/antiox9111098
Agati, G., Stefano, G., Biricolti, S., Tattini, M., 2009: Mesophyll distribution of ‘antioxidant’flavonoid glycosides in Ligustrum vulgare leaves under contrasting sunlight irradiance. Annals of Botany 104(5), 853–861. https://doi.org/10.1093/aob/mcp177
Agati, G., Tattini, M., 2010: Multiple functional roles of flavonoids in photoprotection. New Phytologist 186(4), 786–793. https://doi.org/10.1111/j.1469-8137.2010.03269.x
Ančev, M., Goranova, V., 2006: Trichome morphology of eleven genera of the tribe Alysseae (Brassicaceae) occurring in Bulgaria. Willdenowia 36(1), 193–204.
Bucar, F., Plazibat, M., Knauder, E., Maleš, Ž., 2005: LC/PDA/MS/MS analysis of flavonoids in some taxa of Alyssum and Aurinia (Brassicaceae) from Croatia. In: Zorc, B. (ed.), Book of Abstracts of the Third Croatian Congress on Pharmacy with International Participation, 188–188. Faculty of Pharmacy and Biochemistry, Zagreb.
Damjanović, O., Stevanović, B., 1993: Morpho-anatomical adaptations of endemic species Fibigia triquetra (DC.) Boiss. (Brassicaceae). Bulletin de l’Institut et du Jardin Botaniques de l’Université de Beograd 24-25, 9–19.
Degen, A., 1909: Bemerkungen über einige orientalische Pflanzarten. LIII. Über die Entdeckung eines Vertreters der Gattung Lesquerella im Velebitgebirge. Magyar Botanikai Lapok 8, 3–24
Domac, R., 1993: Degenia Hayek. In: Tutin, T. G., Burges, N. A., Chater A. O., Edmondson J. R., Heywood, V., Moore, D. M., Valentine, D. H., Walters, S. M., Webb, D. A. (eds.), Flora Europaea, 359, vol. 1. 2nd ed. Cambridge University Press, Cambridge.
Dunkić, V., Bezić, N., Mileta, T., 2001: Xeromorphism of trichomes in Lamiaceae species. Acta Botanica Croatica 60(2), 277–283.
Fahn, A., Cutler, F. D., 1992: Xerophytes. Encyclopedia of Plant Anatomy, vol. 3. Gebrüder Bornträger, Berlin, Stuttgart.
Hayek, A., 1910: Die systematische Stellung von Lesquerella velebitica Degen. Oesterreichische botanische Zeitschrift 60, 89–93
Hopewell, T., Selvi, F., Ensikat, H. J., Weigend, M., 2021: Trichome biomineralization and soil chemistry in Brassicaceae from Mediterranean ultramafic and calcareous soils. Plants 10(2), 377. https://doi.org/10.3390/plants10020377
Horvat, I., 1930: Vegetacijske studije o hrvatskim planinama. Zadruge na planinskim goletima [Vegetation studies on Croa-tian mountains. Communities on mountain bare areas]. Rad Jugoslavenske Akademije Znanosti 238, 1–96.
Horvat, I, 1931: Vegetacijske studije o hrvatskim planinama. II. Zadruge na planinskim stijenama i točilima [Vegetation studies on Croatian mountains. II. Communities on mountain rocks and screes]. Rad Jugoslavenske Akademije Znanosti 241, 147–206.
Hutzler, P., Fischbach, R., Heller, W., Jungblut, T., Reuber, S., Schmitz, R., Veit, M., Weissenböck, G., Schnitzler, J., 1998: Tissue localization of phenolic compounds in plants by confocal laser scanning microscopy. Journal of Experimental Botany 49(323), 953–965. https://doi.org/10.1093/jxb/49.323.953
Karabourniotis, G., Liakopoulos, G., Nikolopoulos, D., Bresta, P., 2020: Protective and defensive roles of non-glandular trichomes against multiple stresses: structure–function coordination. Journal of Forestry Research 31(1), 1-12. https://doi.org/10.1007/s11676-019-01034-4
Laoué, J., Fernandez, C., Ormeño, E., 2022: Plant flavonoids in mediterranean species: A focus on flavonols as protective metabolites under climate stress. Plants 11(2), 172. https://doi.org/10.3390/plants11020172
Liakopoulos, G., Stavrianakou, S., Karabourniotis, G., 2006: Trichome layers versus dehaired lamina of Olea europaea leaves: differences in flavonoid distribution, UV-absorbing capacity, and wax yield. Environmental and Experimental Botany 55(3), 294–304. https://doi.org/10.1016/j.envexpbot.2004.11.008
Liakoura, V., Stefanou, M., Manetas, Y., Cholevas, C., Karabourniotis, G., 1997: Trichome density and its UV-B protective potential are affected by shading and leaf position on the canopy. Environmental and Experimental Botany 38(3), 223–229. http://dx.doi.org/10.1016/S0098-8472(97)00005-1
Liber, Z., Surina, B., Nikolić, T., Škrtić, D., Šatović, Z., 2020: Spatial distribution, niche ecology and conservation genetics of Degenia velebitica (Brassicaceae), a narrow endemic species of the north-western Dinaric Alps. Plant Systematics and Evolution 306(4), 64. https://doi.org/10.1007/s00606-020-01695-3
Manetas, Y., 2003: The importance of being hairy: the adverse effects of hair removal on stem photosynthesis of Ver-bascum speciosum are due to solar UV-B radiation. New Phytologist 158, 503–508.
Mastelić, J., Blažević, I., Kosalec, I., 2010: Chemical composition and antimicrobial activity of volatiles from Degenia vele-bitica, a European stenoendemic plant of the Brassicaceae family. Chemistry & Biodiversity, 7(11), 2755–2765. https://doi.org/10.1002/cbdv.201000053
Naumovski, D., 2005: Germination ecology of seeds of endemic species Degenia velebitica (Degen) Hayek (Brassicaceae). Acta Botanica Croatica 64(2), 323–330.
Nikolić, T., Milović, M., Bogdanović, S., Jasprica, N., 2015: Endemi u hrvatskoj flori [Endemics in Croatian flora]. Alfa d.d., Zagreb.
Nikolić, T. (ed.), 2015-2025: Flora Croatica Database. Retrieved April 5, 2025 from http://hirc.botanic.hr/fcd. Pri-rodoslovno-matematički fakultet, Sveučilište u Zagrebu.
Official Gazette (Narodne Novine), 2013: Pravilnik o strogo zaštićenim vrstama [Ordinance of strictly protected species]. 144. Croatian Government, Zagreb.
Rešetnik, I., Satovic, Z., Schneeweiss, G. M., Liber, Z., 2013: Phylogenetic relationships in Brassicaceae tribe Alysseae in-ferred from nuclear ribosomal and chloroplast DNA sequence data. Molecular Phylogenetics and Evolution 69(3), 772–786. https://doi.org/10.1016/j.ympev.2013.06.026
Shen, N., Wang, T., Gan, Q., Liu, S., Wang, L., Jin, B., 2022: Plant flavonoids: Classification, distribution, biosynthesis, and antioxidant activity. Food Chemistry 383, 132531. http://dx.doi.org/10.1016/j.foodchem.2022.132531
Singh, P., Arif, Y., Bajguz, A., Hayat, S., 2021: The role of quercetin in plants. Plant Physiology and Biochemistry, 166, 10–19. http://dx.doi.org/10.1016/j.plaphy.2021.05.023
Skaltsa, H., Verykokidou, E., Harvala, C., Karabourniotis, G., Manetasi, Y., 1994: UV-B protective potential and flavonoid content of leaf hairs of Quercus ilex. Phytochemistry 37(4), 987–990. http://dx.doi.org/10.1016/S0031-9422(00)89514-X
Stamenković, V., Kovačić, S., Juretić, B., Mihelj, D., 2010: Ex-situ protection program for Croatian endemic plant taxa in the Botanical Garden of the Department of Biology, Faculty of Science. In: Jasprica, N., Pandža, M., Milović, M. (eds.), Abstract Book of the Third Croatian Botanical Congress, 180–181. Croatian Botanical Society, Murter, Zagreb.
Stamenković, V., Likić, S., Tkalec, M., 2015: Defence response of Degenia velebitica (Brassicaceae) to solar UV radiation. In: Bogdanović, S., Jogan, N. (eds.), Book of Abstracts of the Sixth Balkan Botanical Congress, 109–110. Natural History Museum Rijeka, Croatian Botanical Society, Botanical Society of Slovenia, University of Rijeka, Rijeka.
Stevanović, B., Vujnović, K., 1990: Morpho‐anatomical adaptations of the endemic species Degenia velebitica (Deg.) Hay. Feddes Repertorium 101(7‐8), 385–389. http://dx.doi.org/10.1002/fedr.19901010710
Sylvester, A. W., Cande, W. Z., Freeling, M., 1991: Developmental morphology of maize leaves using UV light induced autofluorescence of cell wall compounds. Maize Genetics Cooperation Newsletter 65, 34–35.
Tattini, M., Gravano, E., Pinelli, P., Mulinacci, N., Romani, A., 2000: Flavonoids accumulate in leaves and glandular tri-chomes of Phillyrea latifolia exposed to excess solar radiation. New Phytologist 148(1), 69–77. http://dx.doi.org/10.1046/j.1469-8137.2000.00743.x
Tattini, M., Matteini, P., Saracini, E., Traversi, M. L., Giordano, C., Agati, G., 2007: Morphology and biochemistry of non-glandular trichomes in Cistus salvifolius L. leaves growing in extreme habitats of the Mediterranean basin. Plant Biology 9(03), 411–419. http://dx.doi.org/10.1055/s-2006-924662
Tsiftsoglou, O. S., Lazari, D. M., Stefanakis, M. K., Kokkalou, E. L., 2019: Flavonoids and phenolic acids from the aerial parts of Alyssum alyssoides L. (Brassicaceae). Biochemical Systematics and Ecology 83, 51–53. http://dx.doi.org/10.1016/j.bse.2018.12.004
Watkins, J. M., Chapman, J. M., Muday, G. K., 2017: Abscisic acid-induced reactive oxygen species are modulated by fla-vonols to control stomata aperture. Plant Physiology 175(4), 1807–1825. http://dx.doi.org/10.1104/pp.17.01010
Watts, S., Kariyat, R., 2021: Morphological characterization of trichomes shows enormous variation in shape, density and dimensions across the leaves of 14 Solanum species. AoB Plants 13(6), plab071. https://doi.org/10.1093/aobpla/plab071