vuzbiochemiИзвестия вузов. Прикладная химия и биотехнологияProceedings of Universities. Applied Chemistry and Biotechnology2227-29252500-1558ИРНИТУ10.21285/2227-2925-2021-11-2-251-259vuzbiochemi-602Research ArticleФИЗИКО-ХИМИЧЕСКАЯ БИОЛОГИЯPHYSICOCHEMICAL BIOLOGYИсследование роли фоторецепторов у Arabidopsis thaliana в регуляции процесса state transitionsThe role of Arabidopsis thaliana photoreceptors in regulating the process of state transitionsБельковВ. И.BelkovV. I.

Бельков Вадим Игоревич - кандидат биологических наук, младший научный сотрудник.

664033, Иркутск, ул. Лермонтова, 132.

Vadim I. Belkov - Cand. Sci. (Biology), Junior Researcher, Siberian Institute of Plant Physiology and Biochemistry SB RAS.

132, Lermontov St., Irkutsk, 664033.

anvad.irk@rambler.ruБелогубК. Е.BelogubK. E.

Белогуб Кристина Евгеньевна – студентка.

664033, Иркутск, ул. К. Маркса, 1.

Kristina E. Belogub - Student, Irkutsk State University.

1, K. Marx St., Irkutsk, 664033.

kristina200298@yandex.ruГарникЕ. Ю.GarnikE. Yu.

Гарник Елена Юрьевна - кандидат биологических наук, старший научный сотрудник.

664033, Иркутск, ул. Лермонтова, 132.

Elena Yu. Garnik - Cand. Sci. (Biology), Senior Scientist, Siberian Institute of Plant Physiology and Biochemistry SB RAS.

132, Lermontov St., Irkutsk, 664033.

elga74@yandex.ruТарасенкоВ. И.TarasenkoV. I.

Тарасенко Владислав Игоревич – кандидат биологических наук, старший научный сотрудник.

664033, Иркутск, ул. Лермонтова, 132.

Vladislav I. Tarasenko - Cand. Sci. (Biology), Senior Scientist, Siberian Institute of Plant Physiology and Biochemistry SB RAS.

132, Lermontov St., Irkutsk, 664033.

vslav@inbox.ruКонстантиновЮ. М.KonstantinovYu. M.

Константинов Юрий Михайлович – доктор биологических наук, профессор, заведующий лабораторией генетической инженерии растений, СИФИБР СО РАН; профессор, ИГУ.

664033, Иркутск, ул. Лермонтова, 132; 664033, Иркутск, ул. К. Маркса, 1.

Yuri M. Konstantinov - Dr. Sci. (Biology), Professor, Head of the laboratory of Plant Genetic Engineering, Siberian Institute of Plant Physiology and Biochemistry SB RAS; Professor, Irkutsk State University

132, Lermontov St., Irkutsk, 664033; 1, K. Marx St., Irkutsk, 664033.

yukon@sifibr.irk.ruСибирский институт физиологии и биохимии растений СО РАНРоссияSiberian Institute of Plant Physiology and Biochemistry SB RASRussian FederationИркутский государственный университетРоссияIrkutsk State UniversityRussian FederationСибирский институт физиологии и биохимии растений СО РАН; Иркутский государственный университетРоссияSiberian Institute of Plant Physiology and Biochemistry SB RAS; Irkutsk State UniversityRussian Federation202104072021112251259Copyright © Бельков В.И., Белогуб К.Е., Гарник Е.Ю., Тарасенко В.И., Константинов Ю.М., 20212021Бельков В.И., Белогуб К.Е., Гарник Е.Ю., Тарасенко В.И., Константинов Ю.М.Belkov V.I., Belogub K.E., Garnik E.Y., Tarasenko V.I., Konstantinov Y.M.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vuzbiochemi.elpub.ru/jour/article/view/602

Первичное построение фотосинтетического аппарата растений происходит в процессе фотоморфогенеза. Важнейшую роль в инициации и регуляции фотоморфогенеза играют белки-фоторецепторы красного/дальнекрасного (фитохромы) и синего (криптохромы) света. Возбужденные молекулы фитохромов и криптохромов способны взаимодействовать с транскрипционными факторами, изменяя экспрессию ядерных генов, кодирующих белки фотосинтетического аппарата растений. Поскольку свет является непостоянным, вариабельным фактором, растения выработали соответствующие адаптационные механизмы, в частотности, для защиты своего фотосинтетического аппарата. Механизм state transitions обеспечивает быструю адаптацию фотосинтетического аппарата, направленную на повышение эффективности адсорбции света при имеющихся условиях освещенности, и предотвращение усиленной генерации активных форм кислорода в хлоропластах, способной приводить к фотоокислению и даже гибели клеток. Цель данной работы - выявить роль фоторецепторов - фитохромов А и B, а также криптохромов 1 и 2 в регуляции процесса state transitions у модельного растения Arabidopsis thaliana. В качестве исследуемых объектов использовали мутанты арабидопсиса, несущие дефекты по генам фитохромов А и В, криптохромов 1 и 2. В качестве основного метода исследования был использован «голубой нативный» электрофорез в полиакриламидном геле, который позволяет визуализировать state transitions. В ходе исследований было установлено, что данные фоторецепторы не оказывают прямого влияния на редокс-регуляцию механизма state transitions у арабидопсиса. По-видимому, роль этих фоторецепторов в защите фотосинтетического аппарата от избыточной освещенности заключается не в регуляции state transitions, а реализуется более опосредованно, через регуляцию содержания хлорофиллов, каротиноидов и компонентов антиоксидантной системы.

 

The initial formation of the photosynthetic apparatus in plants occurs during photomorphogenesis. The red/far-red (phytochromes) and blue (cryptochrome) light protein-photoreceptors play the most important role in photomorphogenesis initiation and regulation. The exited phytochrome and cryptochrome molecules can interact with transcription factors, changing the expression of nuclear genes, which encode the proteins of the plant photosynthetic apparatus. Since light is a variable factor, plants have developed appropriate adaptation mechanisms, including their photosynthetic apparatus protection. The mechanism of state transitions ensures a rapid adaptation of the photosynthetic apparatus. This adaptation mechanism increases the adsorption efficiency under current light conditions and prevents intensive generation of active forms of oxygen in chloroplasts, which leads to photo-oxidation and even cell death. This work aims to determine the role of photoreceptors - phytochromes A and B, as well as cryptochrome 1 and 2 - in regulating the process of state transitions in the Arabidopsis thaliana model plant. Arabidopsis mutants with the defects on A and B phytochromes and cryptochrome 1 and 2 genes were used as the research objects. The blue native electrophoresis in polyacrylamide gel was used to visualise state transitions. It was found that these photoreceptors had no direct effect on the redox-regulation of the state transitions mechanism in Arabidopsis. Presumably, these photoreceptors protect the photosynthetic apparatus from excessive light not by regulating the state transitions but indirectly, through regulating the chlorophyll, carotenoid and antioxidant components content.

Arabidopsis thalianaфитохромыкриптохромыбелки светособирающего комплексафотосинтетический аппаратArabidopsis thalianaphytochromescryptochromelight-harvesting complex proteinphotosynthetic apparatusРабота выполнена при финансовой поддержке гранта РФФИ № 18-34-00800. В работе использовано оборудование ЦКП «Биоаналитика» СИФИБР СО РАН (г. Иркутск). Авторы благодарны М.В. Кулинченко, к.б.н., старшему научному сотруднику лаборатории генетической инженерии растений СИФИБР СО РАН, за консультации при выполнении методической части работы.This work was supported by the RFBR no. 18-34-00800. Equipment of the Bioanalitika Center for Collective Use, Siberian Institute of Plant Physiology and Biochemistry, Siberian Branch of the Russian Academy of Sciences, Irkutsk, was used in this study. The authors are grateful to Senior Researcher of Plant Genetic Engineering Laboratory, Siberian Institute of Plant Physiology and Biochemistry SB RAS M.V. Koulinchenko for consultations when performing the methodological part of the work.ReferencesHan X., Tohge T., Lalor P., Dockery P., Devaney N., Esteves-Ferreira A.A., et al. Phytochrome A and B regulate primary metabolism in Arabidopsis leaves in response to light // Frontiers in Plant Science. 2017. Vol. 8. P. 1394. https://doi.org/10.3389/fpls.2017.01394Han X, Tohge T, Lalor P, Dockery P, Devaney N, Esteves-Ferreira AA, et al. Phytochrome A and B regulate primary metabolism in Arabidopsis leaves in response to light. Frontiers in Plant Science. 2017;8:1394. https://doi.org/10.3389/fpls.2017.01394Paik I., Huq E. Plant photoreceptors: Multifunctional sensory proteins and their signaling networks // Seminars in Cell and Developmental Biology. 2019. Vol. 92. P. 114-121. https://doi.org/10.1016/j.semcdb.2019.03.007Paik I, Huq E. Plant photoreceptors: Multifunctional sensory proteins and their signaling networks. Seminars in Cell and Developmental Biology. 2019;92:114-121. https://doi.org/10.1016/j.semcdb.2019.03.007Ma D., Li X., Guo Y., Chu J., Fang S., Yan C., et al. Cryptochrome 1 interacts with PIF4 to regulate high temperature-mediated hypocotyl elongation in response to blue light // PNAS. 2016. Vol. 113. Issue 1. P. 224-229. https://doi.org/10.1073/pnas.1511437113Ma D, Li X, Guo Y, Chu J, Fang S, Yan C, et al. Cryptochrome 1 interacts with PIF4 to regulate high temperature-mediated hypocotyl elongation in response to blue light. PNAS. 2016;113(1):224-229. https://doi.org/10.1073/pnas.1511437113Kreslavski V.D., Los D.A., Schmitt F.-J., Zharmukhamedov S.K., Kuznetsov V.V., Allakhverdiev S.I. The impact of the phytochromes on photosynthetic processes // Biochimica et Biophysica Acta (BBA) - Bioenergetics. 2018. Vol. 1859. Issue 5. P. 400-408. https://doi.org/10.1016/j.bbabio.2018.03.003Kreslavski VD, Los DA, Schmitt F-J, Zharmu-khamedov SK, Kuznetsov VV, Allakhverdiev SI. The impact of the phytochromes on photosynthetic processes. Biochimica et Biophysica Acta (BBA) - Bioenergetics. 2018;1859(5):400-408. https://doi.org/10.1016/j.bbabio.2018.03.003Colombo M., Suorsa M., Rossi F., Ferreri R., Tadini L., Barbato R., et al. Photosynthesis control: an underrated short-term regulatory mechanism essential for plant viability // Plant Signaling & Behavior. 2016. Vol. 11. Issue 4. e1165382 (6 p.). https://doi.org/10.1080/15592324.2016.1165382Colombo M, Suorsa M, Rossi F, Ferreri R, Tadini L, Barbato R, et al. Photosynthesis control: an underrated short-term regulatory mechanism essential for plant viability. Plant Signaling & Behavior. 2016;11(4):e1165382 (6 p.). https://doi.org/10.1080/15592324.2016.1165382Dietzel L., Brautigam K., Pfannschmidt T. Photosynthetic acclimation: state transitions and adjustment of photosystem stoichiometry-functional relationships between short-term and long-term light quality acclimation in plants // FEBS Journal. 2008. Vol. 275. Issue 6. P. 1080-1088. https://doi.org/10.1111/j.1742-4658.2008.06264.xDietzel L, Brautigam K, Pfannschmidt T. Photosynthetic acclimation: state transitions and adjustment of photosystem stoichiometry-functional relationships between short-term and long-term light quality acclimation in plants. FEBS Journal. 2008; 275(6):1080-1088. https://doi.org/10.1111/j.1742-4658.2008.06264.xCaffarii S.A., Tibiletti T., Jennings R., Santabarbara S. A comparison between plant photosystem I and photosystem II architecture and functionning // Current Protein and Peptide Science. 2014. Vol. 15. Issue 4. P. 296-331. https://doi.org/10.2174/1389203715666140327102218Caffarii SA, Tibiletti T, Jennings R, Santabarba-ra S. A comparison between plant photosystem I and photosystem II architecture and functioning. Current Protein and Peptide Science. 2014;15(4):296-331. https://doi.org/10.2174/1389203715666140327102218Wunder T., Liu Q., Aseeva E., Bonardi V., Leister D., Pribil M. Control of STN7 transcript abundance and transient STN7 dimerisation are involved in the regulation of STN7 activity // Planta. 2013. Vol. 237. Issue 2. P. 541-558. https://doi.org/10.1007/s00425-012-1775-yWunder T, Liu Q, Aseeva E, Bonardi V, Leister D, Pribil M. Control of STN7 transcript abundance and transient STN7 dimerisation are involved in the regulation of STN7 activity. Planta. 2013;237(2):541-558. https://doi.org/10.1007/s00425-012-1775-yRochaix J.-D., Lemeille S., Shapiguzov A., Samol I., Fucile G., Willig A., et al. Protein kinases and phosphatases involved in the acclimation of the photosynthetic apparatus to a changing light environment // Philosophical Transactions of the Royal Society B. Biological Sciences. 2012. Vol. 367. Issue 1608. P. 3466-3474. https://doi.org/10.1098/rstb.2012.0064Rochaix J-D, Lemeille S, Shapiguzov A, Sa-mol I, Fucile G, Willig A, et al. Protein kinases and phosphatases involved in the acclimation of the photosynthetic apparatus to a changing light environment. Philosophical Transactions of the Royal Society B. Biological Sciences. 2012;367(1608):3466-3474. https://doi.org/10.1098/rstb.2012.0064Verdaguer D., Jansen M.A.K., Llorens L., Morales L.O., Neugart S. UV-A radiation effects on higher plants: exploring the known unknown // Plant Science. 2017. Vol. 255. P. 72-81. https://doi.org/10.1016/j.plantsci.2016.11.014Verdaguer D., Jansen M.A.K., Llorens L., Morales L.O., Neugart S. UV-A radiation effects on higher plants: exploring the known unknown // Plant Science. 2017. Vol. 255. P. 72-81. https://doi.org/10.1016/j.plantsci.2016.11.014Kreslavski V.D., Lyubimov V.Yu., Shirshiko-va G.N., Shmarev A.N., Kosobryukhov A.A., Schmitt F.J., et al. Preillumination of lettuce seedlings with red light enhances the resistance of photosynthetic apparatus to UV-A // Journal of Photochemistry and Photobiology B: Biology. 2013. Vol. 122. P. 1-6. https://doi.org/10.1016/j.jphotobiol.2013.02.016Kreslavski VD, Lyubimov VYu, Shirshikova GN, Shmarev AN, Kosobryukhov AA, Schmitt FJ, et al. Preillumination of lettuce seedlings with red light enhances the resistance of photosynthetic apparatus to UV-A. Journal of Photochemistry and Photobiology B: Biology. 2013;122:1-6. https://doi.org/10.1016/j.jphotobiol.2013.02.016Khudyakova A.Yu., Kreslavski V.D., Shirshikova G.N., Zarmukhamedov S.K., Kosobryukhov A.A., Allakhverdiev S.I. Resistance of Arabidopsis thaliana L. photosynthetic apparatus to UV-B is reduced by deficit of phytochromes B and A // Journal of Photochemistry and Photobiology B: Biology. 2017. Vol. 169. P. 41-46. https://doi.org/10.1016/j.photobiol.2017.02.024Khudyakova AYu, Kreslavski VD, Shirshikova GN, Zarmukhamedov SK, Kosobryukhov AA, Allakhverdiev SI. Resistance of Arabidopsis thaliana L. photosynthetic apparatus to UV-B is reduced by deficit of phytochromes B and A. Journal of Photochemistry and Photobiology B: Biology. 2017;169:41-46. https://doi.org/10.1016/j.photobiol.2017.02.024Aronsson H., Jarvis P. A simple method for isolating import-competent Arabidopsis chloroplasts // FEBS Letters. 2002. Vol. 529. Issue 2-3. P. 215220. https://doi.org/10.1016/s0014-5793(02)03342-2Aronsson H, Jarvis P. A simple method for isolating import-competent Arabidopsis chloroplasts. FEBS Letters. 2002;529(2-3):215-220. https://doi.org/10.1016/s0014-5793(02)03342-2Pribil M., Pesaresi P., Hertle A., Barbato R., Leister D. Role of plastid protein phosphatase TAP38 in LHCII dephosphorylation and thylakoid electron flow // PLoS Biology. 2010. Vol. 8. Issue 1. e1000288 (12 p.). https://doi.org/10.1371/journal.pbio.1000288Pribil M, Pesaresi P, Hertle A, Barbato R, Leister D. Role of plastid protein phosphatase TAP38 in LHCII dephosphorylation and thylakoid electron flow. PLoS Biology. 2010;8(1):e1000288 (12 p.). https://doi.org/10.1371/journal.pbio.1000288Nijtmans G.J., Henderson N.S., Holt I.J. Blue native electrophoresis to study mitochondrial and other protein complexes // Methods. 2002. Vol. 26. Issue 4. P. 327-334. https://doi.org/10.1016/S1046-2023(02)00038-5Nijtmans GJ, Henderson NS, Holt IJ. Blue native electrophoresis to study mitochondrial and other protein complexes. Methods. 2002;26(4):327-334. https://doi.org/10.1016/S1046-2023(02)00038-5Ni Z., Kim E.-D., Ha M., Lackey E., Liu J., Zhang Y., et al. Altered circadian Rhythms regulate growth vigour in hybrids and allopolyploids // Nature. 2009. Vol. 457. Issue 7227. P. 327-331. https://doi.org/10.1038/nature07523Ni Z, Kim E-D, Ha M, Lackey E, Liu J, Zhang Y, et al. Altered circadian Rhythms regulate growth vigour in hybrids and allopolyploids. Nature. 2009;457 (7227):327-331. https://doi.org/10.1038/nature07523Wunder T., Liu Q., Aseeva E., Bonardi V., Leister D., Pribil M. Control of STN7 transcript abundance and transient STN7 dimerisation are involved in the regulation of STN7 activity // Planta. 2013. Vol. 237. P. 541-558. https://doi.org/10.1007/s00425-012-1775-yWunder T, Liu Q, Aseeva E, Bonardi V, Leister D, Pribil M. Control of STN7 transcript abundance and transient STN7 dimerisation are involved in the regulation of STN7 activity. Planta. 2013;237: 541-558. https://doi.org/10.1007/s00425-012-1775-yFey V., Wagner R., Brautigam K., Wirtz M., Hell R., Dietzmann A., et al. Retrograde plastid redox signals in the expression of nuclear genes for chloroplast proteins of Arabidopsis thaliana // The Journal of Biological Chemistry. 2005. Vol. 280. Issue 7. P. 5318-5328. https://doi.org/10.1074/jbc.M406358200Fey V, Wagner R, Brautigam K, Wirtz M, Hell R, Dietzmann A, et al. Retrograde plastid redox signals in the expression of nuclear genes for chloroplast proteins of Arabidopsis thaliana. The Journal of Biological Chemistry. 2005;280(7):5318-5328. https://doi.org/10.1074/jbc.M406358200Carvalho R.F., Campos M.L., Azevedo R.A. The role of phytochrome in stress tolerance // Journal of Integrative Plant Biology. 2011. Vol. 53. Issue 12. P. 920-929. https://doi.org/10.1111/j.1744-79 09.2011.01081.xCarvalho RF, Campos ML, Azevedo RA. The role of phytochrome in stress tolerance. Journal of Integrative Plant Biology. 2011;53(12):920-929. https://doi.org/10.1111/j.1744-7909.2011.01081.xKreslavski V.D., Kosobryukhov A.A., Schmitt F.-J., Semenova G.A., Shirshikova G.N., Khudyako-va A.Y., et al. Photochemical activity and the structure of chloroplasts in Arabidopsis thaliana L. mutants deficient in phytochrome A and B // Protoplasma. 2017. Vol. 254. Issue 3. P. 1283-1293. https://doi.org/10.1007/s00709-016-1020-9Kreslavski VD, Kosobryukhov AA, Schmitt F-J, Semenova GA, Shirshikova GN, Khudyakova AY, et al. Photochemical activity and the structure of chloroplasts in Arabidopsis thaliana L. mutants deficient in phytochrome A and B. Protoplasma. 2017; 254(3):1283-1293. https://doi.org/10.1007/s00709-016-1020-9Casal J.J. Photoreceptor signaling networks in plant responses to shade // Annual Review of Plant Biology. 2013. Vol. 64. P. 403-427. https://doi.org/10.1146/annurev-arplant-050312-120221Casal JJ. Photoreceptor signaling networks in plant responses to shade. Annual Review of Plant Biology. 2013;64:403-427. https://doi.org/10.1146/annurev-arplant-050312-120221Rusaczonek A., Czarnocka W., Kacprzak S., Witon D., Slesak I., Szechynska-Hebda M., et al. Role of phytochromes A and B in the regulation of cell death and acclimatory responses to UV stress in Arabidopsis thaliana // Journal of Experimental Botany. 2015. Vol. 66. Issue 21. P. 6679-6695. https://doi.org/10.1093/jxb/erv375Rusaczonek A, Czarnocka W, Kacprzak S, Witon D, Slesak I, Szechynska-Hebda M, et al. Role of phytochromes A and B in the regulation of cell death and acclimatory responses to UV stress in Arabidopsis thaliana. Journal of Experimental Botany. 2015;66(21):6679-6695. https://doi.org/10.1093/jxb/erv375

The authors declare that there are no conflicts of interest present.