Abstract
Brassinosteroids (BRs), a class of plant steroid hormones, play a significant role in the amelioration of various biotic and abiotic stresses. In order to further explore and elaborate their roles in plants subjected to chilling stress, suspension cultured cells of Chorispora bungeana with or without 24-epibrassinolide (EBR) application were exposed to 4 and 0°C for 5 days. The EBR treated cells exhibited higher viability after exposure to low temperatures compared with the control. Under chilling stress, reactive oxygen species (ROS) levels and lipid peroxidation were increased in the cultured cells, which were significantly inhibited by EBR application. The activities of antioxidative enzymes such as ascorbate peroxidase (APX), catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD) were increased during chilling treatments, and these increases were more significant in the EBR applied suspension cells. The EBR treatment also greatly enhanced contents of ascorbic acid (AsA) and reduced glutathione (GSH) under chilling stress. From these results, it can be concluded that EBR could play the positive roles in the alleviation of oxidative damage caused by ROS overproduction through enhancing antioxidant defense system, resulting in improving the tolerance of C. bungeana suspension cultures to chilling stress.
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Abbreviations
- APX:
-
Ascorbate peroxidase
- AsA:
-
Ascorbic acid or ascorbate
- BRs:
-
Brassinosteroids
- CAT:
-
Catalase
- EBR:
-
24-Epibrassinolide
- GSH:
-
Reduced glutathione
- H2O2 :
-
Hydrogen peroxide
- MDA:
-
Malondialdehyde
- NBT:
-
Nitroblue tetrazolium
- OH− :
-
Hydroxyl radical
- O2 − :
-
Superoxide radical
- POD:
-
Peroxidase
- ROS:
-
Reactive oxygen species
- SOD:
-
Superoxide dismutase
- TBA:
-
Thiobarbituric acid
- TCA:
-
Trichloroacetic acid
- TTC:
-
2, 3, 5-Triphenyltetrazolium chloride
References
Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126
Ali B, Hasan SA, Hayat S, Hayat Q, Yadav S, Fariduddin Q, Ahmad A (2008) A role for brassinosteroids in the amelioration of aluminium stress through antioxidant system in mung bean (Vigna radiata L. Wilczek). Environ Exp Bot 62:153–159. doi:10.1016/j.envexpbot.2007.07.014
Anuradha S, Rao SSR (2001) Effect of brassinosteroids on salinity stress induced inhibition of seed germination and seedlings growth of rice (Oryza sativa L). Plant Growth Regul 33:151–153
Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assay and an assay applicable to acrylamide gels. Anal Biochem 44:276–287
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
Cao S, Xu Q, Cao Y, Qian K, An K, Zhu Y, Binzeng H, Zhao H, Kuai B (2005) Loss of function mutations in DET2 gene lead to an enhanced resistance to oxidative stress in Arabidopsis. Physiol Plant 123:57–66. doi:10.1111/j.1399-3054.2004.00432.x
Chance B, Maehly AC (1955) Assay of catalase and peroxidases. Methods Enzymol 2:764–775
Dhaubhadel S, Chaundhary S, Dobinson KF, Krishna P (1999) Treatment with 24-epibrassinolide, a brassinosteroid, increases the basic thermotolerance of Brassica napus and tomato seedlings. Plant Mol Biol 40:333–342
Ershova A, Khripach V (1996) Effects of epibrassinolide on lipid peroxidation in Pisum sativum at normal aeration and under oxygen deficiency. Russ J Plant Physiol 43:750–752
Fu XY, Chang JF, An LZ, Zhang MX, Xu SJ, Chen T, Liu YH, Xing H, Wang JH (2006) Association of the cold-hardness of Chorispora bungeana with the distribution and accumulation of calcium in the cells and tissues. Environ Exp Bot 55:282–293. doi:10.1016/j.envexpbot.2004.11.009
Griffith OW (1980) Determination of glutathione and glutathione disulphide using glutathione reductase and 2-vinylpyridine. Anal Biochem 106:207–212
Guo FX, Zhang MX, Chen Y, Zhang WH, Xu SJ, Wang JH, An LZ (2006) Relation of several antioxidant enzymes to rapid freezing resistance in suspension cultured cells from alpine Chorispora bungeana. Cryobiology 52:241–250. doi:10.1016/j.cryobiol.2005.12.001
Halliwell B, Grootveld M, Gutteridge JMC (1988) Methods for the measurement of hydroxyl radicals in biochemical system: dioxyribose degradation and aromatic hydroxylation. Methods Biochem Anal 33:59–90
Hariyadi P, Parkin KL (1991) Chilling-induced oxidative stress in cucumber fruits. Postharvest Biol Technol 1:33–45
Hasan SA, Hayat S, Ali B, Ahmad A (2008) 28-Homobrassinolide protects chickpea (Cicer arietinum) from cadmium toxicity by stimulating antioxidants. Environ Pollut 151:60–66. doi:10.1016/j.envpol.2007.03.006
Hidey E, Bjorn LO (1996) Ultraweak light emission, free radicals chilling and light sensitivity. Physiol Plant 98:223–228
Hirai K, Fujii S, Honjo K (1991) The effect of brassinolide on ripening of rice plants under low temperature condition. Jpn J Crop Sci 60(1):29–35
Hodges DM, Delong JM, Forney CF, Prange RK (1999) Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta 207:604–611
Ishikawa M, Robertson AJ, Gusta LV (1995) Comparison of viability tests for assessing cross-adaptation to freezing heat and salt stresses induced by abscisic acid in bromegrass (Bromus inermis Leyss) suspension cultured cells. Plant Sci 107:83–93
Kang YY, Guo SR, Li J, Duan JJ (2007) Effects of 24-epibrassinolide on antioxidant system in cucumber seedling roots under hypoxia stress. China Agric Sci 6(3):281–289
Kim KS, Sa JG (1989) Effects of plant growth regulator, brassinolide, on seedling growth in rice (Oryza sativa L). Res Rep Rural Dev Adm Rice 31(1):49–53
Kuk YI, Shin JS, Burgos NR, Hwang TE, Han O, Cho BH, Jung S, Guh JO (2003) Antioxidative enzymes offer protection from chilling damage in rice plants. Crop Sci 43:2109–2117
Law MY, Charles SA, Halliwell B (1983) Glutathione and ascorbic acid in spinach (Spinacia oleracea) chloroplasts. The effect of hydrogen peroxide and of paraquat. Biochem J 210:899–903
Li L, van Staden J (1998) Effect of plant growth regulators on the antioxidative system in callus of two maize cultivars subjected to water stress. Plant Growth Regul 24:55–66
Mandava NB (1988) Plant growth-promoting brassinosteroids. Ann Rev Plant Physiol Plant Mol Biol 39:23–52
Mazzora LM, Núñez M, Hechararria M, Coll F, Sanchez-Blanco MJ (2002) Influence of brassinosteroids on antioxidant enzyme activity in tomato under different temperatures. Biol Plant 45:593–596
Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplast. Plant Cell Physiol 22:867–880
Nakashita H, Yasuda M, Nitta T, Asami T, Fujikoa S, Arai Y, Sekimata K, Takatsuto S, Yamaguchi I, Yoshida S (2003) Brassinosteroids functions in a broad range of disease resistance in tobacco and rice. Plant J 33:887–898
Noctor G, Foyer CH (1998) Ascorbate and glutathione: keeping active oxygen under control. Annu Rev Plant Physiol Plant Mol Biol 49:249–279
Núñez M, Mazzafera P, Mazzora LM, Siqueira WJ, Zullo MAT (2003) Influence of a brassinosteroid analogue on antioxidant enzymes in rice grown in culture medium with NaCl. Biol Plant 47:67–70
Ogweno JO, Song XS, Shi K, Hu WH, Mao WH, Zhou YH, Yu JQ, Nogués S (2008) Brassinosteroids alleviate heat-induced inhibition of photosynthesis by increasing carboxylation efficiency and enhancing antioxidant systems in Lycopersicon esculentum. J Plant Growth Regul 27:49–57. doi:10.1007/s00344-007-9030-7
Ozdemir F, Bor M, Demiral T, Turkan I (2004) Effects of 24-epibrassinolide on seed germination, seedling growth, lipid peroxidation, proline content and antioxidative system of rice (Oryza sativa L) under salinity stress. Plant Growth Regul 42:203–211
Patterson BD, Mackae EA, Mackae I (1984) Estimation of hydrogen peroxide in plants extracts using titanium (ıv). Anal Biochem 139:487–492
Posmyk MM, Bailly C, Szafrańska K, Jana KM, Corbineau F (2005) Antioxidant enzymes and isoflavonoids in chilled soybean (Glycine max (L.) Merr.) seedling. J Plant Physiol 162:403–412. doi:10.1016/j.jplph.2004.08.004
Sala JM, Lafuente MT (2004) Antioxidant enzymes activities and rindstaining in ‘Naveline’ oranges as affected by storage relative humidity and ethylene conditioning. Postharvest Biol Technol 31:277–285. doi:10.1016/j.postharvbio.2003.10.002
Sasse JM, Smith R, Hudson I (1995) Effect of 24-epibrassinolide on germination of seeds of Eucalyptus camaldulensis in saline conditions. Proc Plant Growth Regul Soc Am 22:136–141
von Tiedemann A (1997) Evidence for a primary role of active oxygen species in induction of host cell death during infection of bean leaves with Botrytis cinerea. Physiol Mol Plant Pathol 50:151–166
Wang B, Zeng G (1993) Effect of epibrassinolide on the resistance of rice seedlings to chilling injury. J Plant Physiol 19:53–60
Wilen RW, Sacco M, Gusta LV, Krishna P (1995) Effects of 24-epibrassinolide on freezing and thermotolerance of bromegrass (Bromus inermis) cell cultures. Physiol Plant 95:195–202
Yu JQ, Zhou YH, Ye SF, Huang LF (2002) 24-Epibrassinolide and abscisic acid protect cucumber seedlings from chilling injury. J Hortic Sci Biotechnol 77:470–473
Zhang TG, Liu YB, Xue LG, Xu SJ, Chen T, Yang TW, Zhang LJ, An LZ (2006) Molecular cloning and characterization of a novel MAP kinase gene in Chorispora bungeana. Plant Physiol Biochem 44:78–84. doi:10.1016/j.plaphy.2006.01.001
Zhao L, He JX, Wang XM, Zhang LX (2008) Nitric oxide protects against polyethyleneglycol-induced oxidative damage in two ecotypes of reed suspension cultures. J Plant Physiol 165:182–191. doi:10.1016/j.jplph.2007.03.002
Zhou BY, Guo ZF, Liu ZL (2005) Effects of abscisic acid on antioxidant systems of Stylosanthe guianensis (Aublet) Sw. under chilling stress. Crop Sci 45:599–605
Acknowledgments
This study has been supported by the National Outstanding Youth Foundation of China (No. 30625008), the National High Technology Research and Development Program of China (863 Program) (No. 2007AA021401) and the National Basic Research Program of China (973 Program) (No. 2007CB108902).
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Liu, Y., Zhao, Z., Si, J. et al. Brassinosteroids alleviate chilling-induced oxidative damage by enhancing antioxidant defense system in suspension cultured cells of Chorispora bungeana . Plant Growth Regul 59, 207–214 (2009). https://doi.org/10.1007/s10725-009-9405-9
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DOI: https://doi.org/10.1007/s10725-009-9405-9