Abstract
Haberlea rhodopensis Friv. is unique with its ability to survive two extreme environmental stresses—desiccation to air-dry state and subzero temperatures. In contrast to desiccation tolerance, the mechanisms of freezing tolerance of resurrection plants are scarcely investigated. In the present study, the role of antioxidant defense in the acquisition of cold acclimation and freezing tolerance in this resurrection plant was investigated comparing the results of two sets of experiments—short term freezing stress after cold acclimation in controlled conditions and long term freezing stress as a part of seasonal temperature fluctuations in an outdoor ex situ experiment. Significant enhancement in flavonoids and anthocyanin content was observed only as a result of freezing-induced desiccation. The total amount of polyphenols increased upon cold acclimation and it was similar to the control in post freezing stress and freezing-induced desiccation. The main role of phenylethanoid glucoside, myconoside and hispidulin 8-C-(2-O-syringoyl-b-glucopyranoside) in cold acclimation and freezing tolerance was elucidated. The treatments under controlled conditions in a growth chamber showed enhancement in antioxidant enzymes activity upon cold acclimation but it declined after subsequent exposure to −10 °C. Although it varied under ex situ conditions, the activity of antioxidant enzymes was high, indicating their important role in overcoming oxidative stress under all treatments. In addition, the activity of specific isoenzymes was upregulated as compared to the control plants, which could be more useful for stress counteraction compared to changes in the total enzyme activity, due to the action of these isoforms in the specific cellular compartments.
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Alscher RG, Erturk N, Heath LS (2002) Role of superoxide dismutases (SODs) in controlling oxidative stress in plants. J Exp Bot 53:1331–1341. https://doi.org/10.1093/jexbot/53.372.1331
Anderson MD, Prasad TK, Stewart CR (1995) Changes in isozyme profiles of catalase, peroxidase, and glutathione reductase during acclimation to chilling in mesocotyls of maize seedlings. Plant Physiol 109:1247–1257. https://doi.org/10.1104/pp.109.4.1247
Anjum NA, Tantray AY, Khan NA, Ahmad A (2020) Reactive oxygen species detection-approaches in plants: insights into genetically encoded FRET-based sensors. J Biotechnol 308:108–117. https://doi.org/10.1016/j.jbiotec.2019.12.003
Arslan Ö, Eyidoğan F, Ekmekçi Y (2018) Freezing tolerance of chickpea: biochemical and molecular changes at vegetative stage. Biol Plant 62:140–148. https://doi.org/10.1007/s10535-017-0760-5
Azevedo RA, Alas RM, Smith RJ, Lea PJ (1998) Response of antioxidant enzymes to transfer from elevated carbon dioxide to air and ozone fumigation, in the leaves and roots of wild-type and catalase-deficient mutant of barley. Physiol Plant 104:280–292. https://doi.org/10.1034/j.1399-3054.1998.1040217.x
Barrero-Gil J, Huertas R, Rambla J, Granell A, Salinas J (2016) Tomato plants increase their tolerance to low temperature in a chilling acclimation process entailing comprehensive transcriptional and metabolic adjustments. Plant Cell Environ 39:2303–2318. https://doi.org/10.1111/pce.12799
Benina M, Obata T, Mehterov N, Ivanov I, Petrov V, Toneva V, Fernie AR, Gechev TS (2013) Comparative metabolic profiling of Haberlea rhodopensis, Thellungiella halophyla, and Arabidopsis thaliana exposed to low temperature. Front Plant Sci 4:499. https://doi.org/10.3389/fpls.2013.00499
Bhatia C, Pandey A, Gaddam SR, Hoecker U, Trivedi PK (2018) Low temperature-enhanced flavonol synthesis requires light-associated regulatory components in Arabidopsis thaliana. Plant Cell Physiol 59:2099–2112. https://doi.org/10.1093/pcp/pcy132
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3
Chandlee JM, Scandalios JG (1983) Gene expression during early kernel developmental in Zea mays. Dev Genet 4:99–115. https://doi.org/10.1002/dvg.1020040205
Daskalova E, Dontcheva S, Yahubyan G, Minkov I, Toneva V (2011) A strategy for conservation and investigation of the protected resurrection plant Haberlea rhodopensis Friv. BioRisk 6:41–60. https://doi.org/10.3897/biorisk.6.1568
Dinakar C, Bartels D (2013) Desiccation tolerance in resurrection plants: new insights from transcriptome, proteome and metabolome analysis. Front Plant Sci 4:482. https://doi.org/10.3389/fpls.2013.00482
Farrant JM, Vander Willigen C, Loffell DA, Bartsch S, Whittaker A (2003) An investigation into the role of light during desiccation of three angiosperm resurrection plants. Plant Cell Environ 26:1275–1286. https://doi.org/10.1046/j.0016-8025.2003.01052.x
Farrant JM (2007) Mechanisms of desiccation tolerance in angiosperm resurrection plants. In: Jenks MA, Wood AJ (eds) Plant desiccation tolerance. Blackwell Publishing Ltd, Oxford, pp 51–90
Farrant JM, Moore JP (2011) Programming desiccation-tolerance: from plants to seeds to resurrection plants. Curr Opin Plant Biol 14:340–345. https://doi.org/10.1016/j.pbi.2011.03.018
Feierabend J (2005) Catalases in plants: molecular and functional properties and role in stress defense. In: Smirnoff N (ed) Antioxidants and reactive oxygen species in plants. Blackwell Publishing Ltd, Oxford, pp 101–140
Fernández-Marín B, Neuner G, Kuprian E, Laza JM, García-Plazaola JI, Verhoeven A (2018) First evidence of freezing tolerance in a resurrection plant: insights into molecular mobility and zeaxanthin synthesis in the dark. Physiol Plant 163:472–489. https://doi.org/10.1111/ppl.12694
Fernández-Marín B, Nadal M, Gago J, Fernie AR, López-Pozo M, Artetxe U, García-Plazaola JI, Verhoeven A (2020) Born to revive: molecular and physiological mechanisms of double tolerance in a paleotropical and resurrection plant. New Phytol 226:741–759. https://doi.org/10.1111/nph.16464
Foyer CH, Noctor G (2005) Oxidant and antioxidant signaling in plants, a re-evaluation of the concept of oxidative stress in a physiological context. Plant Cell Environ 28:1056–1071. https://doi.org/10.1111/j.1365-3040.2005.01327.x
Georgieva K, Röding A, Büchel C (2009) Changes in some thylakoid membrane proteins and pigments upon desiccation of the resurrection plant Haberlea rhodopensis. J Plant Physiol 166:1520–1528. https://doi.org/10.1016/j.jplph.2009.03.010
Georgieva K, Dagnon S, Gesheva E, Bojilov D, Mihailova G, Doncheva S (2017) Antioxidant defense during desiccation of the resurrection plant Haberlea rhodopensis. Plant Physiol Biochem 114:51–59. https://doi.org/10.1016/j.plaphy.2017.02.021
Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930. https://doi.org/10.1016/j.plaphy.2010.08.016
Gołębiowska-Pikania G, Kopeć P, Surówka E, Krzewska M, Dubas E, Nowicka A, Rapacz M, Wójcik-Jagła M, Malaga S, Żur I (2017) Changes in protein abundance and activity involved in freezing tolerance acquisition in winter barley (Hordeum vulgare L.). J Proteom 169:58–72. https://doi.org/10.1016/j.jprot.2017.08.019
Gould KS, Markham KR, Smith RH, Goris JJ (2000) Functional role of anthocyanins in the leaves of Quintinia serrata A. Cunn J Exp Bot 51:1107–1115. https://doi.org/10.1093/jexbot/51.347.1107
Guo Z, Ou W, Lu S, Zhong Q (2006) Differential responses of antioxidative system to chilling and drought in four rice cultivars differing in sensitivity. Plant Physiol Biochem 44:828–836. https://doi.org/10.1016/j.plaphy.2006.10.024
Hughes NM (2011) Winter leaf reddening in evergreen species. New Phytol 190:573–581. https://doi.org/10.1111/j.1469-8137.2011.03662.x
Janda T, Szalai G, Rios-Gonzalez K, Veisz O, Páldi E (2003) Comparative study of frost tolerance and antioxidant activity in cereals. Plant Sci 164:301–306. https://doi.org/10.1016/S0168-9452(02)00414-4
Janda T, Majláth I, Szalai G (2014) Interaction of temperature and light in the development of freezing tolerance in plants. J Plant Growth Regul 33:460–469. https://doi.org/10.1007/s00344-013-9381-1
Kappen L (1966) Der Einfluß des Wassergehaltes auf die Widerstandsfähigkeit von Pflanzen gegenüber hohen und tiefen Temperaturen, untersucht an Blättern einiger Farne und von Ramonda myconi. Flora Allg Bot Ztg Abt A Physiol Bioch 156:427–445. https://doi.org/10.1016/S0367-1836(17)30278-1
Kolupaev YE, Ryabchun NI, Vayner AA, Yastreb TO, Oboznyi AI (2015) Antioxidant enzyme activity and osmolyte content in winter cereal seedlings under hardening and cryostress. Russ J Plant Physiol 62:499–506. https://doi.org/10.1134/S1021443715030115
Kondeva-Burdina M, Zheleva-Dimitrova D, Nedialkov P, Girreser U, Mitcheva M (2013) Cytoprotective and antioxidant effects of phenolic compounds from Haberlea rhodopensis Friv. (Gesneriaceae). Pharmacognosy Mag 9:294–301. https://doi.org/10.4103/0973-1296.117822
Koonjul PK, Brandt WF, Lindsey GG, Farrant JM (2000) Isolation and characterisation of chloroplasts from Myrothamnus flabellifolius Welw. J Plant Physiol 156:584–594. https://doi.org/10.1016/S0176-1617(00)80217-5
Kosová K, Vítámvás P, Prášil IT (2007) The role of dehydrins in plant response to cold. Biol Plant 51:601–617. https://doi.org/10.1007/s10535-007-0133-6
Kovi M, Ergon A, Rognli O (2016) Freezing tolerance revisited-effects of variable temperatures on gene regulation in temperate grasses and legumes. Curr Opin Plant Biol 33:140–146. https://doi.org/10.1016/j.pbi.2016.07.006
Król A, Amarowicz R, Weidner S (2015) The effects of cold stress on the phenolic compounds and antioxidant capacity of grapevine (Vitis vinifera L.) leaves. J Plant Physiol 189:97–104. https://doi.org/10.1016/j.jplph.2015.10.002
Kumar S, Asif MH, Chakrabarty D, Tripathi RD, Dubey RS, Trivedi PK (2013) Expression of a rice Lambda class of glutathione S-transferase, OsGSTL2, in Arabidopsis provides tolerance to heavy metal and other abiotic stresses. J Hazard Mater 248:228–237. https://doi.org/10.1016/j.jhazmat.2013.01.004
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685. https://doi.org/10.1038/227680a0
Lenné T, Bryant G, Hocart CH, Huang CX, Ball MC (2010) Freeze avoidance: a dehydrating moss gathers no ice. Plant Cell Environ 33:1731–1741. https://doi.org/10.1111/j.1365-3040.2010.02178.x
Li Z, Wakao S, Fischer BB, Niyogi KK (2009) Sensing and responding to excess light. Ann Rev Plant Biol 60:239–260. https://doi.org/10.1146/annurev.arplant.58.032806.103844
Mihailova G, Petkova S, Georgieva K (2009) Changes in some antioxidant enzyme activities in Haberlea rhodopensis during desiccation at high temperature. Biotechnol Biotec Eq 23:561–564. https://doi.org/10.1080/13102818.2009.10818487
Mihailova G, Solti Á, Sárvári É, Keresztes Á, Rapparini F, Velitchkova M, Simova-Stoilova L, Aleksandrov V, Georgieva K (2020) Freezing tolerance of photosynthetic apparatus in the homoiochlorophyllous resurrection plant Haberlea rhodopensis. Environ Exp Bot 178:104157. https://doi.org/10.1016/j.envexpbot.2020.104157
Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410. https://doi.org/10.1016/S1360-1385(02)02312-9
Mittova V, Volokita M, Guy M, Tal M (2000) Activities of SOD and the ascorbate-glutathione cycle enzymes in subcellular compartments in leaves and roots of the cultivated tomato and its wild salt-tolerant relative Lycopersicon pennellii. Physiol Plant 110:42–51. https://doi.org/10.1034/j.1399-3054.2000.110106.x
Mladenov P, Zasheva D, Djilianov D, Tchorbadjieva M (2015) Towards proteomics of desiccation tolerance in the resurrection plant Haberlea rhodopensis. C R Acad Bulg Sci 68:59–64
Moore JP, Farrant JM, Lindsey GG, Brandt WF (2005) The South African and Namibian populations of the resurrection plant Myrothamnus flabellifolius are genetically distinct and display variation in their galloylquinic acid composition. J Chem Ecol 31:2823–2834. https://doi.org/10.1007/s10886-005-8396-x
Moore JP, Le NT, Brandt WF, Driouich A, Farrant JM (2009) Towards a systems-based understanding of plant desiccation tolerance. Trends Plant Sci 14:110–117. https://doi.org/10.1016/j.tplants.2008.11.007
Morse M, Rafudeen MS, Farrant JM (2011) An overview of the current understanding of desiccation tolerance in the vegetative tissues of higher plants. Adv Bot Res 57:319–347. https://doi.org/10.1016/B978-0-12-387692-8.00009-6
Moyankova D, Mladenov P, Berkov S, Peshev D, Georgieva D, Djilianov D (2014) Metabolic profiling of the resurrection plant Haberlea rhodopensis during desiccation and recovery. Physiol Plant 152:675–687. https://doi.org/10.1111/ppl.12212
Mundree SG, Baker B, Mowla S, Peters S, Marais S, Vander Willigen C, Govender K, Maredza A, Muyanga S, Farrant JM, Thomson JA (2002) Physiological and molecular insights into drought tolerance. Afr J Biotechnol 1:28–38. https://doi.org/10.5897/AJB2002.000-006
Murray JR, Hackett WP (1991) Dihydroflavonol reductase activity in relation to differential anthocyanin accumulation in juvenile and mature phase Hedera helix L. Plant Physiol 97:343–351. https://doi.org/10.1104/pp.97.1.343
Noctor G, Foyer CH (1998) Ascorbate and glutathione: keeping active oxygen under control. Ann Rev Plant Physiol Plant Mol Biol 49:249–279. https://doi.org/10.1146/annurev.arplant.49.1.249
Ouellet F, Charron JB (2013) Cold acclimation and freezing tolerance in plants. John Wiley & Sons Ltd, Chichester, pp 1–10
Posmyk MM, Bailly C, Szafrańska K, Janas KM, Corbineau F (2005) Antioxidant enzymes and isoflavonoids in chilled soybean (Glycine max (L.) Merr.) seedlings. J Plant Physiol 162:403–412. https://doi.org/10.1016/j.jplph.2004.08.004
Rakić T, Lazarević M, Jovanović ŽS, Radović S, Siljak-Yakovlev S, Stevanović B, Stevanović V (2014) Resurrection plants of the genus Ramonda: prospective survival strategies–unlock further capacity of adaptation, or embark on the path of evolution? Front Plant Sci 4:550. https://doi.org/10.3389/fpls.2013.00550
Ricci G, Bello ML, Caccuri AM, Galiazzo F, Federici G (1984) Detection of glutathione transferase activity on polyacrylamide gels. Anal Biochem 143:226–230. https://doi.org/10.1016/0003-2697(84)90657-2
Rudikovskaya EG, Fedorova GA, Dudareva LV, Makarova LE, Rudikovskii AV (2008) Effect of growth temperature on the composition of phenols in pea roots. Russ J Plant Physiol 55:712–715. https://doi.org/10.1134/S1021443708050178
Ruelland E, Vaultier M-N, Zachowski A, Hurry V (2009) Cold signaling and cold acclimation in plants. Adv Bot Res 49:35–150. https://doi.org/10.1016/S0065-2296(08)00602-2
Russell R, Lei T, Nilsen E (2009) Freezing induced leaf movements and their potential implications to early spring carbon gain: Rhododendron maximum as exemplar. Funct Ecol 23:463–471. https://doi.org/10.1111/j.1365-2435.2008.01534.x
Schulz E, Tohge T, Zuther E, Fernie AR, Hincha DK (2015) Natural variation in flavonol and anthocyanin metabolism during cold acclimation in Arabidopsis thaliana accessions. Plant Cell Environ 38:1658–1672. https://doi.org/10.1111/pce.12518
Schulz E, Tohge T, Zuther E, Fernie AR, Hincha DK (2016) Flavonoids are determinants of freezing tolerance and cold acclimation in Arabidopsis thaliana. Sci Rep 6:34027. https://doi.org/10.1038/srep34027
Sutinen ML, Arora R, Wisniewski M, Ashworth E, Strimbeck R, Palta J (2001) Mechanisms of frost survival and freeze-damage in nature. In: Bigras FJ, Colombo SJ (eds) Conifer cold hardiness, Tree Physiology, vol 1. Springer, Dordrecht, pp 89–120
Suzuki N, Mittler R (2006) Reactive oxygen species and temperature stresses: a delicate balance between signaling and destruction. Physiol Plant 126:45–51. https://doi.org/10.1111/j.0031-9317.2005.00582.x
Thomashow MF (1998) Role of cold-responsive genes in plant freezing tolerance. Plant Physiol 118:1–8. https://doi.org/10.1104/pp.118.1.1
Thomashow MF (1999) Plant cold acclimation: freezing tolerance genes and regulatory mechanisms. Ann Rev Plant Biol 50:571–599. https://doi.org/10.1146/annurev.arplant.50.1.571
Thomashow MF (2010) Molecular basis of plant cold acclimation: insights gained from studying the CBF cold response pathway. Plant Physiol 154:571–577. https://doi.org/10.1104/pp.110.161794
Veljovic-Jovanovic S, Kukavica B, Navari-Izzo F (2008) Characterization of polyphenol oxidase changes induced by desiccation of Ramonda serbica leaves. Physiol Plant 132:407–416. https://doi.org/10.1111/j.1399-3054.2007.01040.x
Verhoeven AS, Swanberg A, Thao M, Whiteman J (2005) Seasonal changes in leaf antioxidant systems and xanthophyll cycle characteristics in Taxus x media growing in sun and shade environments. Physiol Plant 123:428–434. https://doi.org/10.1111/j.1399-3054.2005.00471.x
Verhoeven A, García-Plazaola JI, Fernández-Marín B (2018) Shared mechanisms of photoprotection in photosynthetic organisms tolerant to desiccation or to low temperature. Environ Exp Bot 154:66–79. https://doi.org/10.1016/j.envexpbot.2017.09.012
Wang X, Peng Y, Singer JW, Fessehaie A, Krebs SL, Arora R (2009) Seasonal changes in photosynthesis, antioxidant systems and ELIP expression in a thermonastic and non-thermonastic Rhododendron species: a comparison of photoprotective strategies in overwintering plants. Plant Sci 177:607–617. https://doi.org/10.1016/j.plantsci.2009.08.009
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K.G. designed the research, performed statistical data analysis and wrote the manuscript with input from other authors. G.M. was responsible for plant treatments and sampling. L.S. and L.G. did protein extraction and analysis of antioxidant enzymes activity. S.D. measured the changes in polyphenols and M.V. determined the content of antocyanins and flavonoids. All authors analyzed the data, revised and edited the manuscript.
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Georgieva, K., Mihailova, G., Gigova, L. et al. The role of antioxidant defense in freezing tolerance of resurrection plant Haberlea rhodopensis. Physiol Mol Biol Plants 27, 1119–1133 (2021). https://doi.org/10.1007/s12298-021-00998-0
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DOI: https://doi.org/10.1007/s12298-021-00998-0