Abstract
A simple method set for assessing biochemical changes associated with osmotic stress responses was developed using coffee (Coffea arabica L.) leaf disks. Stress was induced by polyethylene glycol (PEG) exposure. Quantitative evaluation of tissue physiological stress parameters was carried out using analytical methods to validate the conversion of classic qualitative histochemical tests for localizing lipid peroxidation, hydrogen peroxide, and total xanthine alkaloids into semi-quantitative assays. Relative electrolyte leakage (EL%) and chlorophyll content (SPAD index) were also recorded. EL% levels of treated disks were higher than those of control ones, whereas SPAD indexes were comparable. Histochemical localization indicated that levels of lipid peroxidation, H2O2, and total xanthines were also higher under osmotic stress than in control conditions. Semi-quantitative data obtained by image processing of histochemical staining consistently matched quantitative evaluations. Chromatographic analyses revealed that theophylline and caffeine concentrations increased in the presence of PEG, whereas theobromine remained constant in relation to the control. The methods herein described can be useful to rapidly acquire initial data regarding biochemical osmotic stress responses in coffee tissues based on simple staining and imaging steps. Moreover, it is likely that the same method may be applicable to other types of stresses and plant species upon minor adjustments.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data Availability
The data supporting the findings of this study are available within the article and/or its supplementary materials.
Abbreviations
- FAO:
-
Food and Agriculture Organization
- ICO:
-
International Coffee Organization
- EL:
-
Electrolyte leakage
- EL%:
-
Relative electrolyte leakage
- MS:
-
Culture medium (Murashige and Skoog culture medium)
- PEG:
-
Polyethylene glycol 6000
- H2O2 :
-
Hydrogen peroxide
- SA:
-
Stained area
- Ci:
-
Initial electrical conductivity
- Cf:
-
Final electrical conductivity
- SPAD:
-
Soil Plant Analysis Development (chlorophyll index)
- TCA:
-
Trichloroacetic acid
- TBA:
-
Thiobarbituric acid
- MDA:
-
Malondialdehyde
- FW:
-
Fresh weight
- HPLC:
-
High-performance liquid chromatography
- TB:
-
Theobromine
- TP:
-
Theophylline
- CF:
-
Caffeine
- ROS:
-
Reactive oxygen species
References
Alexieva V, Sergiev I, Mapelli S, Karanov E (2001) The effect of drought and ultraviolet radiation on growth and stress markers in pea and wheat. Plant Cell Environ 24:1337–1344. https://doi.org/10.1046/j.1365-3040.2001.00778.x
Awasthi JP, Saha B, Chowardhara B, Devi SS, Borgohain P, Panda SK (2018) Qualitative analysis of lipid peroxidation in plants under multiple stresses through Schiff’s reagent: a histochemical approach. BioProtoc 8(8):e2807. https://doi.org/10.21769/BioProtoc.2807
Bajji M, Kinet J, Lutts S (2002) The use of the electrolyte leakage method for assessing cell membrane stability as a water stress tolerance test in durum wheat. Plant Growth Reg 36:61–70. https://doi.org/10.1023/A:1014732714549
Bruggink H, Kraak HL, Dijkema MHGE, Bekendam J (2008) Some factors influencing electrolyte leakage from maize (Zea mays L.) kernels. Seed Sci Res 1:15–20. https://doi.org/10.1017/S0960258500000581
Choinski JS, Gould KS (2010) Immature leaves of Weinmannia racemosa are more heat tolerant than mature leaves based on differences in chlorophyll a fluorescence and solute leakage. N Z J Bot 48(3):163–177. https://doi.org/10.1080/0028825X.2010.505945
Chowdhury SR, Maleque M, Shihan MH (2012) Development and validation of a simple RP-HPLC method for determination of caffeine in pharmaceutical dosage forms. Asian J Pharm Ana 2:1–4. https://doi.org/10.5958/2231-5675
DaMatta FM (2004) Ecophysiological constraints on the production of shaded and unshaded coffee: a review. Field Crops Res 86:99–114. https://doi.org/10.1016/j.fcr.2003.09.001
DaMatta FM, Ramalho JDC (2006) Impacts of drought and temperature stress on coffee physiology and production: a review. Braz J Plant Physiol 18:55–81. https://doi.org/10.1590/S1677-04202006000100006
Demidchik V, Straltsova D, Medvedev SS, Pozhvanov GA, Sokolik A, Yurin V (2014) Stress-induced electrolyte leakage: the role of K+-permeable channels and involvement in programmed cell death and metabolic adjustment. J Exp Bot 65:1259–1270. https://doi.org/10.1093/jxb/eru004
Eira MTS, Silva EAA, Castro RD, Dussert S, Walters C, Bewley JD, Hilhost HWM (2006) Coffee seed physiology. Braz J Plant Physiol 18:149–163. https://doi.org/10.1590/S1677-04202006000100011
Emanuil N, Akram MS, Ali S, Majrashi A, Iqbal M, El-Esawi MA, Ditta A, Alharby HF (2022) Exogenous caffeine (1,3,7-trimethylxanthine) application diminishes cadmium toxicity by modulating physio-biochemical attributes and improving the growth of Spinach (Spinacia oleracea L.). Sustainability 14(2806):1–19. https://doi.org/10.3390/su14052806
Fahad S, Bajwa AA, Nazir U, Anjum SA, Farooq A, Zohaib A, Sadia S, Nasim W, Adkins S, Saud S, Ihsan MZ, Alharby H, Wu C, Wang D, Huang J (2017) Crop production under drought and heat stress: plant responses and management options. Front Plant Sci 29:1–16. https://doi.org/10.3389/fpls.2017.01147
FAO, 2023. Coffee. https://www.fao.org/markets-and-trade/commodities/coffee/en/. Accessed 23 Jun 2023.
Fernandes CN (2011). Identification and validation of differentially expressed genes related to drought response in Siriema Coffea arabica plants. Dissertation. University of Lavras (Brazil). http://repositorio.ufla.br/. Accessed 16 May 2023
Frischknecht PM, Baumann TW (1985) Stress induced formation of purine alkaloids in plant tissue culture of Coffea arabica. Phytochem 24(10):2255–2257. https://doi.org/10.1016/S0031-9422(00)83020-4
Gong M, Chen B, Li Z, Guo L (2001) Heat-shock-induced cross adaptation to heat, chilling, drought and salt stress in maize seedlings and involvement of H2O2. J Plant Physiol 158:1125–1130. https://doi.org/10.1078/0176-1617-00327
Guimarães SF (2015). Physiological responses in postharvest of basil leaves (Ocimum basilicum L.). Dissertation, Federal University of Viçosa, Minas Gerais (Brazil). https://locus.ufv.br//handle/123456789/6636. Accessed 10 Apr 2023
Guo R, Hao W, Gong D (2012) Effects of water stress on germination and growth of linseed seedlings (Linum usitatissimum L), photosynthetic efficiency and accumulation of metabolites. J Agric Sci 4(10):253–265. https://doi.org/10.1186/s40064-016-2348-5
Hniličková H, Hnilička F, Orsák M, Hejnák V (2019) Effect of salt stress on growth, electrolyte leakage, Na+ and K+ content in selected plant species. Plant Soil Environ 65(2):90–96. https://doi.org/10.17221/620/2018-PSE
ICO, 2023. Trade statistics tables. http://www.ico.org/trade_statistics.asp?section=Statistics. Accessed 23 Jun 2023.
Kinghorn AD, Falk H, Gibbons S, Kobayashi J (2017) Progress in the chemistry of organic natural products. Springer, Switzerland
Kumar A, Naik GK, Simmi PSS, Giridhar P (2015) Salinity and drought response alleviate caffeine content of young leaves of Coffea canephora var Robusta cv S274. J Appl Biol Biotechnol 3(3):50–60. https://doi.org/10.7324/JABB.2015.3310
Lindén L, (2002). Measuring cold hardiness in woody plants. Dissertation. University of Helsinki (Finland). https://helda.helsinki.fi/. Accessed 22 May 2023
Liu J, Wen B, Liu X, Yang Y, Li M, Wang X (2022) Molecular and metabolic changes under environmental stresses: the biosynthesis of quality components in preharvest tea shoots. Horticulturae 8(173):1–20. https://doi.org/10.3390/horticulturae8020173
Matsuura HN, Rau MR, Fett-Neto AG (2014) Oxidative stress and production of bioactive monoterpene indole alkaloids: biotechnological implications. Biotechnol Lett 36:191–200. https://doi.org/10.1007/s10529-013-1348-6
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Sano H, Kim Y, Choi Y (2013) Like cures like: caffeine immunizes plants against biotic stresses. Adv Bot Res 68:273–300. https://doi.org/10.1016/B978-0-12-408061-4.00010-9
Shi J, Fu XZ, Peng T, Huang XS, Fan QJ, Liu JH (2010) Spermine pretreatment confers dehydration tolerance of citrus in vitro plants via modulation of antioxidative capacity and stomatal response. Tree Physiol 30(7):914–922. https://doi.org/10.1093/treephys/tpq030
Smirnoff N, Arnaud D (2019) Hydrogen peroxide metabolism and functions in plant. New Phytol 221(3):1173–1664. https://doi.org/10.1111/nph.15488
Velikova V, Yordanov I, Edreva A (2000) Oxidative stress and some antioxidant systems in acid rain-treated bean plants: protective role of exogenous polyamines. Plant Sci 151:59–66. https://doi.org/10.1016/S0168-9452(99)00197-1
Vilasboa J, Costa CT, Ransan LG, Mariath JEA, Fett-Neto AG (2022) Microcutting redox profile and anatomy in Eucalyptus spp. with distinct adventitious rooting competence. Front Plant Sci 11:1–12. https://doi.org/10.3389/fpls.2020.620832
Watanabe S, Matsumoto M, Hakomori Y, Takagi H, Shimada H, Sakamoto A (2014) The purine metabolite allantoin enhances abiotic stress tolerance through synergistic activation of abscisic acid metabolism. Plant, Cell Environ 37:1022–1036. https://doi.org/10.1111/pce.12218
Whitlow TH, Bassuk NL, Ranney TG, Reichert DL (1992) An improved method for using electrolyte leakage to assess membrane competence in plant tissues. Plant Physiol 98:198–205. https://doi.org/10.1104/pp.98.1.198
Yadav V, Arif N, Singh VP, Guerriero G, Berni R, Shinde S, Raturi G, Deshmukh R, Sandalio LM, Chauhan DK, Tripathi DK (2021) Histochemical techniques in plant science: more than meets the eye. Plant Cell Physiol 62(10):1509–1527. https://doi.org/10.1093/pcp/pcab022
Zwiewka M, Nodzyński T, Robert S, Vanneste S, Friml J (2015) Osmotic stress modulates the balance between exocytosis and clathrin-mediated endocytosis in Arabidopsis thaliana. Mol Plant 8(8):1175–1187. https://doi.org/10.1016/j.molp.2015.03.007
Acknowledgements
We thank Professor Dr. Jorge Ernesto de Araújo Mariath (Plant Anatomy Laboratory-LAVeg, UFRGS) for access to the stereoscopic microscope used in the analyses.
Funding
This work was supported by the Research Support Foundation of the State of Rio Grande do Sul (FAPERGS, Project 19/2551–0001709-0), the National Council for Scientific and Technological Development (CNPq, Project 310775/2021–3), the Coordination for the Improvement of Higher Education Personnel (CAPES, Finance code 001), and the Organization of American States (OAS).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing interests.
Additional information
Handling Editor: Peter Nick
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
De Palma, N., Fett-Neto, A.G. A semi-quantitative histochemical method for assessment of biochemical responses to osmotic stress in Coffea arabica leaf disks. Protoplasma (2024). https://doi.org/10.1007/s00709-024-01941-2
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00709-024-01941-2