Skip to main content
SpringerLink
Log in

Brassinosteroids alleviate chilling-induced oxidative damage by enhancing antioxidant defense system in suspension cultured cells of Chorispora bungeana

  • Original Paper
  • Published:
Plant Growth Regulation Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

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

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assay and an assay applicable to acrylamide gels. Anal Biochem 44:276–287

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • Chance B, Maehly AC (1955) Assay of catalase and peroxidases. Methods Enzymol 2:764–775

    Article  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • 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

    CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • Griffith OW (1980) Determination of glutathione and glutathione disulphide using glutathione reductase and 2-vinylpyridine. Anal Biochem 106:207–212

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Hariyadi P, Parkin KL (1991) Chilling-induced oxidative stress in cucumber fruits. Postharvest Biol Technol 1:33–45

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Hidey E, Bjorn LO (1996) Ultraweak light emission, free radicals chilling and light sensitivity. Physiol Plant 98:223–228

    Article  Google Scholar 

  • 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

    CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    CAS  Google Scholar 

  • 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

    Google Scholar 

  • 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

    CAS  Google Scholar 

  • 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

    CAS  PubMed  Google Scholar 

  • 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

    Article  Google Scholar 

  • Mandava NB (1988) Plant growth-promoting brassinosteroids. Ann Rev Plant Physiol Plant Mol Biol 39:23–52

    Article  CAS  Google Scholar 

  • 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

    Article  Google Scholar 

  • Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplast. Plant Cell Physiol 22:867–880

    CAS  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Noctor G, Foyer CH (1998) Ascorbate and glutathione: keeping active oxygen under control. Annu Rev Plant Physiol Plant Mol Biol 49:249–279

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  Google Scholar 

  • Patterson BD, Mackae EA, Mackae I (1984) Estimation of hydrogen peroxide in plants extracts using titanium (ıv). Anal Biochem 139:487–492

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • Wang B, Zeng G (1993) Effect of epibrassinolide on the resistance of rice seedlings to chilling injury. J Plant Physiol 19:53–60

    Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    CAS  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

Download references

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).

Author information

Authors and Affiliations

  1. State Key Laboratory of Arid Agroecology, Lanzhou University, 730000, Lanzhou, People’s Republic of China

    Yajie Liu, Zhiguang Zhao, Jing Si, Cuixia Di, Jin Han & Lizhe An

Authors
  1. Yajie Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhiguang Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jing Si
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Cuixia Di
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jin Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Lizhe An
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lizhe An.

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10725-009-9405-9

Keywords

Search

Navigation