PLANT BIOLOGY
ANIMAL BIOLOGY
SUBSCRIPTION
E-SUBSCRIPTION
 
MAP
MAIN PAGE

 

 

 

 

Malyukova L.S., Nechaeva T.L., Zubova M.Yu., Gvasalia M.V., Koninskaya N.G., Zagoskina N.V. Physiological and biochemical characterization of tea (Camellia sinensis L.) microshoots in vitro: the norm, osmotic stress, and effects of calcium. Sel'skokhozyaistvennaya Biologiya [Agricultural Biology], 2020, Vol. 55, № 5, p. 970-980.
(how to cite this article)

doi: 10.15389/agrobiology.2020.5.970eng

UDC: 633.72:581.1:58.085

Supported financially from the Russian Foundation for Basic Research and the Administration of the Krasnodar Territory, grant No. 19-416-230049, and from the Ministry of Education and Science of the Russian Federation within the framework of the state tasks of the FRC SSC RAS No. 0683-2019-0003 and the Timiryazev Institute of Plant Physiology RAS No. AAAA-A-19-11904189005-8

 

PHYSIOLOGICAL AND BIOCHEMICAL CHARACTERIZATION OF TEA (Camellia sinensis L.) MICROSHOOTS in vitro: THE NORM, OSMOTIC STRESS, AND EFFECTS OF CALCIUM

L.S. Malyukova1 ✉, T.L. Nechaeva2, M.Yu. Zubova2,
M.V. Gvasalia1, N.G. Koninskaya1, N.V. Zagoskina2 ✉

1Federal Research Centre the Subtropical Scientific Centre RAS, 2/28, ul. Yana Fabriciusa, Sochi, 354002 Russia, e-mail MalukovaLS@mail.ru (corresponding author ✉), m.v.gvasaliya@mail.ru, natakoninskaya@mail.ru;
2Timiryazev Institute of Plant Physiology RAS, 35, ul. Botanicheskaya, Moscow, 127276 Russia, e-mail nechaevatatyana.07@yandex.ru, mariaz1809@gmail.com, nzagoskina@mail.ru (corresponding author ✉)

ORCID:
Malyukova L.S. orcid.org/0000-0003-1531-5745
Gvasalia M.V. orcid.org/0000-0001-7394-4377
Nechaeva T.L. orcid.org/0000-0003-3341-4763
Koninskaya N.G. orcid.org/0000-0002-2126-5863
Zubova M.Yu. orcid.org/0000-0001-7704-8537
Zagoskina N.V. orcid.org/0000-0002-1457-9450

Received June 10, 2020

 

Stress tolerance is an important trait, that determines the productivity of plants under drought, hypothermia, mineral deficiency, and salinity. Numerous studies of various agricultural crops (J.K. Zhu, 2016; E. Fleta-Soriano, S. Munné-Bosch, 2016), including tea crop (Camellia sinensis L.), were aimed at solving this problem due to the global aridization of the climate. (T.K. Maritim et al., 2015; L.S. Samarina et al., 2019). Along with the sufficiently detailed physiological, biochemical and molecular studies of tea drought tolerance, the exogenous regulation of tolerance by using of chemical and biological substances is still not investigated. In addition, the important role of calcium ions (Ca2+) in the cell recognition of an external stressor by the triggering signal transduction has been shown in many crops (M.C. Kim, 2009; E.G. Rikhvanov et al., 2014). In these studies, tissue culture media supplemented with the osmotically active substances (R.M. Pérez-Clemente et al., 2012; M.K. Rai et al., 2011) and artificial biosystems (microshoots and tissues in vitro), are often used as “drought models” to reveal cellular adaptation mechanisms. However, just a few studies were conducted aimed at deciphering the biochemical and molecular responses of tea plant to stress using tissue culture tool (L.S. Samarina et al., 2018; M.V. Gvasaliya et al., 2019). In this article, for the first time, we investigated the role of calcium in plant adaptation to long-term osmotic stress based on earlier published protocols of tea tissue culture (M.V. Gvasaliya, 2013) and osmotic stress induction protocols.  We also demonstrated the prospect of studying the role of exogenous inducers in increasing plant tolerance using “drought models”. This work aimed to identify the effect of different concentrations of calcium (Ca2+) in the culture medium on the functional state of tea microshoots grown under mannitol-induced osmotic stress in vitro comparing with control. The changes in morphophysiological state of the leaves, leaves water content, cells membrane permeability, malondialdehyde, proline, and photosynthetic pigments were analyzed. It was found that increased Ca2+content in the nutrient medium (from 440 to 880 mg/l) resulted the slower leaves development and significant decrease of malondialdehyde and cell membranes permeability of tea microshoots (by 50 %, р ≤ 0.05) during the long-term cultivation of tea microshoots in vitro (4 months), indicating inhibition of lipid peroxidation processes. The addition of mannitol (40 g/l) to the culture medium reduced the water content of the shoots (on average by 2 %, р ≤ 0.05), thereby forming light osmotic stress, which led to the accumulation of proline (an increase of 30-40 %, р ≤ 0.05), as well as to the structural and functional rearrangement of the photosynthetic apparatus (a decrease in the amount of photosynthetic pigments by an average of 35-40 %). In addition, a significant decrease of malondialdehyde (by 50-70 %, p ≤ 0.05) and the intensity of electrolyte leakage from leaf tissues (on average by 50 %, p ≤ 0.05) were observed, indicating a less pronounced oxidative stress in comparison with control (without mannitol). An increase in the Ca2+ concentration in the nutrient medium (from 440 to 880 mg/l) (in the presence of mannitol) did not significantly affect the water content in the leaves and the photosynthetic apparatus (content and ratio of chlorophylls/carotenoids). An insignificant effect of calcium (in the presence of mannitol) manifested itself in a significant decrease in malondialdehyde by 20 μmol/g dry weight. Consequently, the increased concentration of calcium (660-880 mg/l) in the nutrient medium provides an improvement in the functional state of long-term cultivated tea microshoots in vitro (4 months) by reducing the activity of lipid peroxidation in membranes and increasing their stability. The revealed patterns confirm the positive role of calcium ions in the reduction of combined oxidative stress caused by long-term cultivation of plants in vitro in combination with osmotic stress.

Keywords: tea plants, Camellia sinensis L., in vitro microshoots, calcium, mannitol, osmotic stress, pigments, proline, malondialdehyde.

 

REFERENCES

  1. Zhu J.K. Abiotic stress signaling and responses in plants. Cell, 2016, 167(2): 313-324 CrossRef
  2. Fleta-Soriano E., Munné-Bosch S. Stress memory and the inevitable effects of drought: a physiological perspective. Frontiers in Plant Science, 2016, 7: 143 CrossRef
  3. Marcińska I., Czyczyło-Mysza I., Skrzypek E., Filek M., Grzesiak S., Grzesiak M.T., Janowiak F., Hura T., Dziurka M., Dziurka K., Nowakowska A., Quarrie S.A. Impact of osmotic stress on physiological and biochemical characteristics in drought-susceptible and drought-resistant wheat genotypes. Acta Physiologiae Plantarum, 2013, 35(2): 451-461 CrossRef
  4. Samarina L.S., Ryndin A.V., Malyukova L.S., Gvasaliya M.V., Malyarovskaya V.I. Physiological mechanisms and genetic factors of the tea plant Camellia sinensis (L.) Kuntze response to drought (review). Sel'skokhozyaistvennaya biologiya [Agricultural Biology], 2019, 54(3): 458-468 CrossRef
  5. Fayez K.A., Bazaid S.A. Improving drought and salinity tolerance in barley by application of salicylic acid and potassium nitrate. Journal of the Saudi Society of Agricultural Sciences, 2014, 13(1): 45-55 CrossRef
  6. Yang Y., Guo Y. Unraveling salt stress signaling in plants. Journal of Integrative Plant Biology, 2018, 60(9): 796-804 CrossRef
  7. Medvedev S.S. Fiziologiya rastenii, 2005, 52(2): 282-305 (in Russ.).
  8. Song W.Y., Zhang Z.B., Shao H.B., Guo X.L., Cao H.X., Zhao H.B., Fu Z.Y., Hu X.J. Relationship between calcium decoding elements and plant abiotic-stress resistance. International Journal of Biological Sciences, 2008, 4(2): 116-125 CrossRef
  9. Maritim T.K., Kamunya S.M., Mireji P., Wendia C.M., Muoki R.C., Cheruiyot E.K., Wachira F.N. Physiological and biochemical response of tea (Camellia sinensis (L.) O. Kuntze) to water-deficit stress. The Journal of Horticultural Science and Biotechnology, 2015, 90(4): 395-400 CrossRef
  10. Hetherington A.M., Brownlee C. The generation of Ca2+ signals in plants. Annual Review of Plant Biology, 2004, 55: 401-427 CrossRef 
  11. Kim M.C. Calcium and calmodulin-mediated regulation of gene expression in plant. Molecular Plant, 2009, 2(1): 13-21 CrossRef
  12. Saidi Y., Finka A., Muriset M., Bromberg Z., Weiss Y. G., Maathuis F.J., Goloubinoff P. The heat shock response in moss plants is regulated by specific calcium-permeable channels in the plasma membrane. Plant Cell, 2009, 21: 2829-2843 CrossRef
  13. Rikhvanov E.G., Fedoseeva I.V., Pyatrikas D.V., Borovskii G.B., Voinikov V.K. Fiziologiya rastenii, 2014, 61(2): 155-169 CrossRef (in Russ.).
  14. Shu M.Y., Fan M.Q. Effect of osmotic stress and calcium on membrane-lipid peroxidation and the activity of defense enzymes in fir seedling. Forest Research, 2000, 4: 391-396.
  15. Upadhyaya H., Panda S.K., Dutta B.K. CaCl2 improves post-drought recovery potential in Camellia sinensis (L) O. Kuntze. Plant Cell Reports, 2011, 30(4): 495-503 CrossRef 
  16. Bhagat R.M., Baruah R.D., Cacigue S. Climate and tea [Camellia sinensis (L.) O. Kuntze] production with special reference to north eastern India: a review. Journal of Environmental Research and Development, 2010, 4(4): 1017-1028.
  17. Malyukova L.S., Kozlova N.V., Rogozhina E.V., Strukova D.V., Kerimzade V.V., Velikii A.V. Cultivating subtropical crops on the Black Sea coast of Russia: ecological and agrochemical aspects. Sel'skokhozyaistvennaya biologiya [Agricultural Biology], 2014, 3: 24-31 CrossRef (in Russ.).
  18. Baruah R.D., Bhagat R.M. Climate trends of Northeastern India: a longterm pragmatic analysis for tea production. Two and a Bud, 2012, 59(2): 46-49.
  19. Osmolovskaya N., Shumilina J., Kim A., Didio A., Grishina T., Bilova T., Frolov A. Methodology of drought stress research: experimental setup and physiological characterization. International Journal of Molecular Sciences, 2018, 19(12): 4089-4114 CrossRef
  20. Rai M.K., Kalia R.K., Singh R., Gangola M.P., Dhawan A.K. Developing stress tolerant plants through in vitro selection — an overview of the recent progress. Environmental and Experimental Botany, 2011, 71(1): 89-98 CrossRef
  21. Pérez-Clemente R.M., Gómez-Cadenas A. In vitro tissue culture, a tool for the study and breeding of plants subjected to abiotic stress conditions. In: Recent advances in plant in vitro culture. A. Leva, L.M.R. Rinaldi (eds.). IntechOpen Limited, London, 2012: 91-108 CrossRef
  22. Sunaina N.A., Singh N.B. PEG imposed water deficit and physiological alterations in hydroponic cabbage. Iranian Journal of Plant Physiology, 2016, 6(2): 1651-1658.
  23. Gelmesa D., Dechassa N., Mohammed W., Gebre E., Monneveux P., Bündig C., Winkelmann T. In vitro screening of potato genotypes for osmotic stress tolerance. Open Agriculture, 2017, 2(1): 308-316 CrossRef
  24. Piwowarczyk B., Kamińska I., Rybiński W. Influence of PEG generated osmotic stress on shoot regeneration and some biochemical parameters in Lathyrus culture. Czech Journal of Genetics and Plant Breeding, 2014, 50(2): 77-83 CrossRef
  25. Abu-Romman S., Suwwan M., Al-Shadiadeh A., Hasan H. Effects of osmotic stress on cucumber (Cucumis sativus l.) microshoots cultured on proliferation medium. World Applied Sciences Journal, 2012, 20(2): 177-181 CrossRef
  26. Tejavathi D.H., Devaraj V.R., Murthy S.M., Nijagunaiah R., Shobha K. Effect of PEG induced osmotic stress on proline, protein and relative water content in vitro plants of Macrotyloma uniflorum (Lam.) Verdc. Acta Hortic., 2010, 865: 87-93 CrossRef
  27. Gvasaliya M.V., Samarina L.S., Malyukova L.S., Malyarovskaya V.I., Rakhmangulov R.S., Koninskaya N.G., Platonova N.B., Pashchenko O.I. Vestnik Michurinskogo gosudarstvennogo agrarnogo universiteta, 2019, 4(59): 49-53 (in Russ.).
  28. Gvasaliya M.V. Novye tekhnologii, 2020, 3: 117-124 CrossRef (in Russ.). 
  29. Gvasaliya M.V. Sadovodstvo i vinogradarstvo, 2013, 4: 20-22 (in Russ.).
  30. Zubova M.Yu., Nikolaeva T.N., Nechaeva T.L., Malyukova L.S., Zagoskina N.V. Khimiya rastitel'nogo syr'ya, 2019, 4: 249-257 CrossRef (in Russ.).
  31. Yoshida K., Matsuo K. A simple method of evaluating the freezing resistance of tea plants (Camellia sinensis (L.) Kuntze) by measuring electrolyte leakage from low-temperature-treated overwintering buds and leaves. Chagyo Kenkyu Hokoku (Tea Research Journal), 2012, 113: 63-69 CrossRef
  32. Tsypurskaya E.V., Kazantseva V.V., Fesenko A.N., Zagoskina N.V. Growth of buckwheat (Fagopyrum esculentum Moench) seed-lings and the accumulation of primary and secondary metabolites under various mineral nutrition conditions. Sel'skokhozyaistvennaya biologiya [Agricultural Biology], 2019, 54(5): 946-957 CrossRef
  33. Shlyk A.A. V sbornike: Biokhimicheskie metody v fiziologii rastenii [In: Biochemical methods in plant physiology]. Moscow, 1971: 154-170 (in Russ.).
  34. Tholakalabavi A., Zwiazek J.J., Thorpe T.A. Effect of mannitol and glucose-induced osmotic stress on growth, water relations, and solute composition of cell suspension cultures of poplar (Populus deltoides var. occidentalis) in relation to anthocyanin accumulation. In Vitro Cellular & Developmental Biology-Plant, 1994, 30(3): 164-170 CrossRef
  35. Xu C., Li X., Zhang L. The effect of calcium chloride on growth, photosynthesis, and antioxidant responses of Zoysia japonica under drought conditions. PloS ONE, 2013, 8(7): e68214 CrossRef
  36. Li Z., Tan X.F., Lu K., Liu Z.M., Wu L.L. The effect of CaCl2 on calcium content, photosynthesis, and chlorophyll fluorescence of tung tree seedlings under drought conditions. Photosynthetica, 2017, 55(3): 553-560 CrossRef
  37. Kaczmarek M., Fedorowicz-Stronska O., Głowacka K., Waskiewicz A., Sadowski J.  CaCl2 treatment im-proves drought stress tolerance in barley (Hordeum vulgare L.). Acta Physiologiae Plantarum, 2017, 39(1): 41-52 CrossRef
  38. Farmer E.E., Mueller M.J. ROS-mediated lipid peroxidation and RES-activated signaling. Annual Review of Plant Biology, 2013, 64: 429-450 CrossRef
  39. Noctor G., Mhamdi A., Foyer C.H. The roles of reactive oxygen metabolism in drought: not so cut and dried. Plant Physiology, 2014, 164(4): 1636-1648 CrossRef
  40. Madany M., Khalil R. Seed priming with ascorbic acid or calcium chloride mitigates the adverse effects of drought stress in sunflower (Helianthus annuus L.) seedlings. The Egyptian Journal of Experimental Biology (Botany), 2017, 13(1): 119-133 CrossRef
  41. Demidchik V., Straltsova D., Medvedev S.S., Pozhvanov G.A., Sokolik A., Yurin V. Stress-induced electrolyte leakage: the role of K+-permeable channels and involvement in programmed cell death and metabolic adjustment. Journal of Experimental Botany, 2014, 65(5): 1259-1270 CrossRef
  42. Hu W., Tian S.B., Di Q., Duan S.H., Dai K. Effects of exogenous calcium on mesophyll cell ultrastructure, gas exchange, and photosystem II in tobacco (Nicotiana tabacum Linn.) under drought stress. Photosynthetica, 2018, 56(4): 1204-1211 CrossRef
  43. Sofronova V.E., Chepalov V.A., Dymova O.V., Golovko T.K. Fiziologiya rastenii, 2014, 61(2): 266-274 CrossRef (in Russ.).
  44. Fathi A., Tari D.B. Effect of drought stress and its mechanism in plant. International Journal of Life Sciences, 2016, 10(1): 1-6 CrossRef
  45. Sun T., Yuan H., Cao H., Yazdani M., Tadmor Y., Li L. Carotenoid metabolism in plants: the role of plastids. Molecular Plant, 2018, 11(1): 58-74 CrossRef

 

back

 


CONTENTS

 

 

Full article PDF (Rus)

Full article PDF (Eng)