Determination of lipid ordering in chloroplast microdomains of wheat leaves under soil drought conditions

The degree of lipid matrix ordering in the chloroplast microdomains of leaves from two wheat cultivars – a drought-tolerant Buryatskaya Ostistaya and a moisture-sensitive Diamant 2 – was studied under control conditions and under prolonged water deficit (10 days). Lipid ordering was assessed based on generalized polarization values obtained using the Laurdan lipophilic probe. Stress intensity was evaluated according to the level of reactive oxygen species. Under control conditions, both cultivars showed an increase in the generalized polarization values in rafts of zone 1 (R1). In contrast, in rafts of zone 2 (R2), this parameter remained unchanged in the drought-tolerant cultivar, whereas in the moisture-sensitive cultivar it decreased on days 4 and 10 relative to day 1. Under drought conditions, the generalized polarization values in R1 of the tolerant cultivar decreased on days 4 and 10. A decrease in generalized polarization was also observed in R2, with the most pronounced reduction occurring on day 4. An increase in the generalized polarization values was observed in the microdomains of both zones in Diamant 2, except on day 7, when a decrease in microviscosity was recorded. Hence, the microviscosity changes detected in the lipids of the studied microdomains, under both control and stress conditions, suggest that chloroplast membrane microdomains in wheat leaves may not only be involved in cellular responses to stress, but may also participate in other physiological and biochemical processes.

1. Plotnikova L.Ya., Glushakov D.A., Yusov V.S. Results of the study of drought resistance of durum wheat and its components in Western Siberia. Vestnik of Omsk SAU. 2022;4(48):56-70. (In Russian). DOI: 10.48136/2222-0364_2022_4_56. EDN: KRNNCU.

2. Huseynova I.M., Suleymanov S.Y., Rustamova S.M., Aliyev J.A. Drought-induced changes in photosynthetic membranes of two wheat (Triticum aestivum L.) cultivars. Biochemistry (Moscow). 2009;74(8):903-909. DOI: 10.1134/S0006297909080124. EDN: XMCBUB.

3. Valitova J.N., Khabibrakhmanova V.R., Belkina A.V., Renkova A.G., Minibayeva F.V. The lipid profile of wheat roots treated with membranotropic agents. Biologicheskie membrany. 2020;37(6):466-476. (In Russian). DOI: 10.31857/S0233475520060080. EDN: EIFYJM.

4. Zhukov A., Popov V. Eukaryotic cell membranes: structure, composition, research methods and computational modelling. International Journal of Molecular Sciences. 2023;24(13):11226. DOI: 10.3390/ijms241311226.

5. Nurminsky V.N., Nesterov V.N., Rosentsvet O.A., Rakevich A.L., Bukin Yu.S., Kapustina I.S., Ozolina N.V. Analysis of lipid order of raft structures in mitochondrial membranes of halophites with the aid of fluorescent microscopy. Biologicheskie membrany. 2021;38(4):306-314. (In Russian). DOI: 10.31857/S0233475521040083.

6. Regen S.L. The Origin of lipid rafts. Biochemistry. 2020;59(49):4617-4621. DOI: 10.1021/acs.biochem.0c00851.

7. Radyukhin V.A., Baratova L.A. Molecular mechanisms of raft organization in biological membranes. Russian Journal of Bioorganic Chemistry. 2020;46(3):269-279. DOI: 10.1134/S1068162020030164. EDN: ICMPEN.

8. Petrov N.Yu., Onishchenko N.S. Photosynthetic activity and productivity of winter wheat varieties depending on applied biological products. Bulletin of Altai State Agricultural University. 2012;10:23-25. (In Russian). EDN: PDFOST.

9. Gajate C., Mollinedo F. Lipid raft isolation by sucrose gradient centrifugation and visualization of raft-located proteins by fluorescence microscopy: the use of combined techniques to assess Fas/CD95 location in rafts during apoptosis triggering. In: Bieberich E. (ed.). Lipid Rafts. Methods in Molecular Biology. New York: Humana; 2021, vol. 2187, p. 147-186. DOI: 10.1007/978-1-0716-0814-2_9.

10. Kapustina I.S., Gurina V.V., Spiridonova E.V., Ozolina N.V. Characteristics of membrane structures of wheat chloroplasts. Biologicheskie membrany. 2023;40(6):472-478. (In Russian). DOI: 10.31857/S0233475523060051. EDN: FMXCTT.

11. Rozentsvet O.A., Bogdanova E.S., Nesterov V.N., Kosobryukhov A.A. Daytime dynamics of the photosynthetic apparatus’ structural and functional parameters in wild halophytes. Russian Journal of Plant Physiology. 2019;66(6):901-910. DOI: 10.1134/S1021443719060104. EDN: FNDLNG.

12. Korsukova A.V., Grabelnych O.I., Pobezhimova T.P., Koroleva N.A., Fedoseeva I.V., Pavlovskaya N.S., et al. Cold hardening prevents H₂O₂-induced programmed cell death in maize coleoptiles. Journal of Stress Physiology & Biochemistry. 2013;9(1):246-257. EDN: PNGTDX.

13. Bradford M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry. 1976;72:248-254. DOI: 10.1016/0003-2697(76)90527-3.

14. Belugin B.V., Zhestkova I.M., Trofimova M.S. Affinity of PIP-aquaporins to sterol-enriched domains in plasma membrane of the cells of etiolated pea seedlings. Biologicheskie membrany. 2010;27(5):394-403. (In Russian). EDN: MVSKSF.

15. Glantz S.A. Primer of biostatistics; 1992, 472 p. (Russ. ed.: Mediko-biologicheskaya statistika. Moscow: Praktika; 1999, 462 p.).

16. Yurina N.P., Odintsova M.S. Chloroplast retrograde signaling system. Russian Journal of Plant Physiology. 2019;66(4):509-520. DOI: 10.1134/S1021443719040149. EDN: IATOLZ.

17. Osipova S.V., Ostrovskaya R.M., Tret’yakova A.V. Genetic aspects of drought resistance in polyploid plants by the example of wheat Triticum aestivum L. Russian Journal of Plant Physiology. 2022;69(3):44. DOI: 10.1134/S1021443722020145. EDN: QJKNKC.

18. Kuryanchyk T.G., Kozel N.V. Photosynthetic apparatus of barley plants treated with 5-aminolevulinic acid: mechanisms of adaptation to drought. Experimental Biology and Biotechnology. 2022;3:26-38. (In Russian). DOI: 10.33581/2957-5060-2022-3-26-38. EDN: EKULWY.

19. Nesterenko T.V., Tikhomirov A.A., Shikhov V.N. Induction of chlorophyll fluorescence and assessment of plant resistance to adverse effects. Zhurnal obshchei biologii. 2007;68(6):444-458. (In Russian). EDN: IBGYDH.

20. Vershubskii A.V., Tikhonov A.N. Structural and functional aspects of electron transport thermoregulation and ATP synthesis in chloroplasts. Biochemistry (Moscow). 2021;86(1):92-104. DOI: 10.1134/S0006297921010090. EDN: KFCDOC.

21. Kabashnikova L.F. Photosynthetic apparatus and stress in plants. Minsk: Belorusskaya nauka; 2014, 272 p. (In Russian). EDN: UGOKDB.

22. Maslova T.G., Markovskaya E.F., Slemnev N.N. Functions of carotenoids in leaves of higher plants (an overview). Zhurnal obshchei biologii.2020;81(4):297-310. (In Russian). DOI: 10.31857/S0044459620040065. EDN: XSLDGB.

23. Szilágyi А., Selstam E., Åkerlund H.-E. Laurdan fluorescence spectroscopy in the thylakoid bilayer: the effect of violaxanthin to zeaxanthin conversion on the galactolipid dominated lipid environment. Biochimica et Biophysica Acta (BBA) – Biomembranes. 2008;1778(1):348-355. DOI: 10.1016/j.bbamem.2007.10.006.

24. Zhukov A.V. On qualitative composition of membrane lipids in plant cells. Russian Journal of Plant Physiology. 2021;68(2):367-383. DOI: 10.31857/S001533032101022X. EDN: UDXOWC.

25. Rozentsvet O.A., Bogdanova E.S., Nurminsky V.N., Nesterov V.N., Chernyshov M.Yu. Detergent-resistant membranes in chloroplasts and mitochondria of the halophyte salicornia perennans under salt stress. Plants. 2023;12(6):1265. DOI: 10.3390/plants12061265.