Skip to main content
SpringerLink
Log in

In vitro rice shoot apices as simple model to study the effect of NaCl and the potential of exogenous proline and glutathione in mitigating salinity stress

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

Abstract

This study was conducted using in vitro rice shoot apices cultures of two Malaysian rice cultivars—MR 220 and MR 253 to investigate the effect of NaCl and the exogenous application of proline and glutathione in mitigating the salt-induced damages. The results showed that high NaCl concentrations (150, 200, 250 and 300) mM significantly impeded plant growth resulted in reduction in plant height, root length, biomass and chlorophyll content. Results showed that the supplementation of proline and glutathione effectively ameliorates salt stress induced damages. The plant height recorded at 150 mM NaCl (control) were 9.8 and 10.3 cm for MR 220 and MR 253. With the supplementation of 5 mM proline, the plant height of MR 220 and MR 253 increased to 14.8 and 15.0 cm respectively. Similarly, the plant height of MR 220 and MR 253 was further increased to 20.3 and 21.3 cm when 10 mM glutathione was added exogenously. Fresh weight was recorded as 0.06 g in 150 mM NaCl media for both cultivars and increased to 0.13, 0.26, 0.16, 0.23 g (MR 220) and 0.11, 0.14, 0.27, 0.32 (MR 253) with the supplementation of 5, 10, 15 and 20 mM proline respectively. It was noted that supplementation of 5 mM proline successfully increase the endogenous proline content from 9.05 to 58.4 and 15.8 to 70.5 μmol/g for both MR 220 and MR 253 respectively. In addition, supplementation of 5 and 10 mM glutathione increased chlorophyll content from 7.0 mg/g (NaCl) for both cultivars to 13.09, 17.06 mg/g for MR 220 and 14.7, 12.6 mg/g for MR 253 respectively. These results highlighted the potential role of exogenously applied proline and glutathione in mitigating the detrimental effect of salt 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.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Abogadallah GM (2010) Antioxidative defense under salt stress. Plant Signal Behav 5:369–374

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Abu-Romman S, Suwwan M (2008) Influence of NaCl salinity on growth and physiology of cucumber microshoots grown on rooting medium. Dirasat: Agric Sci 35:73–80

    Google Scholar 

  • Ahmad P, Prasad MNV (2012) Abiotic stress responses in plants: metabolism, productivity and sustainability. Springer Science Business Media LLC, New York

    Book  Google Scholar 

  • Ahmad P, Sharma S (2008) Salt stress and phyto-biochemical responses of plants. Plant Soil Environ 54:89–99

    Google Scholar 

  • Ahmad MSA, Javed F, Ashraf M (2007) Iso-osmotic effect of NaCl and PEG on growth, cations and free proline accumulation in callus tissue of two indica rice (Oryza sativa. L) genotypes. Plant Growth Regul 53:53–63

    Article  Google Scholar 

  • Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress and signal transduction. Annu Rev Plant Biol 55:373–399

    Article  CAS  PubMed  Google Scholar 

  • Arnon DI (1949) Copper enzymes in isolated chloroplasts Polyphenoloxidase in Beta vulgaris. Plant Physiol 24:1–15

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Asch F, Dingkuhn M, Sow A, Audebert A (2005) Drought-induced changes in rooting patterns and assimilate partitioning between root and shoot in upland rice. Field Crop Res 93:223–236

    Article  Google Scholar 

  • Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205–207

    Article  CAS  Google Scholar 

  • Benderradji L, Brini F, Kellou K, Ykhelf N, Djekoun A, Masmoudi K, Bouzerour H (2012) Callus induction, proliferation, and plantlets regeneration of two bread wheat (Triticum aestivum L.) genotypes under saline and heat stress conditions. ISRN Agron Article ID 367851

  • Cano EA, Perez A, Moreno V, Caro M, Bolarin M (1998) Evaluation of salt tolerance in cultivated and wild tomato species through in vitro shoot apex culture. Plant Cell Tissue Organ Cult 53(1):19–26

    Article  Google Scholar 

  • Cao F, Liu L, Ibrahim W, Cai Y, Wu F (2013) Alleviating effects of exogenous glutathione, glycinebetaine, brassinosteroids and salicylic acid on cadmium toxicity in rice seedlings (Oryza sativa). Agrotechnol 2:1

    Article  Google Scholar 

  • Chaves MM, Flexas J, Pinheiro C (2009) Photosynthesis under drought and salt stress: regulation mechanisms from whole plant to cell. Ann Bot 103:551–560

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Chutipaijit S, Cha-um S, Sompornpailin K (2011) High contents of proline and anthocyanin increase protective response to salinity in Oryza sativa L. spp. indica. Aus J crop Sci 5(10):1191–1198

    CAS  Google Scholar 

  • Deivanai S, Xavier R, Vinod V, Timalata K, Lim OF (2011) Role of exogenous proline in ameliorating salt stress at early stage in two rice cultivars. J Stress Physiol Biochem 7(4):157–174

    Google Scholar 

  • Ehsanpour AA, Fatahian N (2003) Effects of salt and proline on Medicago sativa callus. Plant Cell Tissue Organ Cult 73:53–56

    Article  CAS  Google Scholar 

  • Fadzilla NM, Finch RP, Burdon RH (1997) Salinity, oxidative stress and antioxidant responses in shoot cultures of rice. J Exp Bot 48:325–331

    Article  CAS  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–168

    Article  CAS  PubMed  Google Scholar 

  • Gossett DR, Banks SW, Millhollon EP, Lucas MC (1996) Antioxidant response to NaCl stress in a control and a NaCl-tolerant cotton cell line grown in the presence of paraquat, buthionine sulfoximine and exogenous glutathione. Plant Physiol 112:803–809

    PubMed Central  PubMed  Google Scholar 

  • Hare PD, Cress WA, van Staden J (2002) Disruptive effects of exogenous proline on chloroplast and mitochondrial ultrastructure in Arabidopsis leaves. S Afr J Bot 68:393–396

    CAS  Google Scholar 

  • Hasanuzzaman M, Nahar K, Fujita M (2013) Plant response to salt stress and role of exogenous protectants to mitigate salt-induced damages. In: Ahmad P, Azooz MM, Prasad MNV (eds) Ecophysiology and Responses of Plants under Salt Stress. Springer, New York, pp 25–87

    Chapter  Google Scholar 

  • Hasanuzzaman M, Alam MM, Rahan A, Hasanuzzaman M, Nahar K. Fujita M (2014) Exogenous proline and glycine betaine mediated upregulation of antioxidant defense and glyoxylase systems provides better protection against salt-induced oxidative stress in two rice (Oryza sativa L.) varieties. Biomed Res Int 2014, Article ID 757219

  • Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophy 125:189–198

    Article  CAS  Google Scholar 

  • Heyneke E, Luschin-Ebengreuth N, Krajcer I, Wolkinger V, Müller M, Zechmann B (2013) Dynamic compartment specific changes in glutathione and ascorbate levels in Arabidopsis plants exposed to different light intensities. BMC Plant Biol 13:104

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Hmida-Sayari A, Gargouri-Bouzid R, Bidani A, Jaoua L, Savoure A, Jaoua S (2005) Overexpression of D1-pyrroline-5-carboxylate synthetase increases proline production and confers salt tolerance in transgenic potato plants. Plant Sci 169:746–752

    Article  CAS  Google Scholar 

  • Horie T, Karahana I, Katsuhara M (2012) Salinity tolerance mechanisms in glycophytes: an overview with the central focus on rice plants. Rice 5(1):11

    Article  Google Scholar 

  • Jose MP, Francisco JQ (2002) Plants and sodium ions: keeping company with the enemy. Genome Biol 3(6):1017.1–1017.4

    Google Scholar 

  • Kattab H (2007) Role of glutathione and polyadenylic acid on the oxidative defense systems of two cultivars of canola seedlings grown under saline condition. Aust J Basic Appl Sci 1:323–334

    CAS  Google Scholar 

  • Kavi Kishor PB, Sangam S, Amrutha RN, Laxmi PS, Naidu NR, Rao KRSS, Rao Sreenath, Reddy KJ, Theriappan P, Sreenivasulu N (2005) Regulation of proline biosynthesis, degradation, uptake and transport in higher plants: its implications in plant growth and abiotic stress tolerance. Curr Sci 88:424–438

    Google Scholar 

  • Kaya C, Tuna AL, Dikilitas M, Cullu MA (2010) Responses of some enzymes and key growth parameters of salt stressed maize plants to foliar and seed applications of kinetin and indole acetic acid. J Plant Nutr 59:1–6

    Google Scholar 

  • Khush GS (2005) What it will take to feed 5.0 billion rice consumers in 2030? Plant Mol Biol 59:1–6

    Article  CAS  PubMed  Google Scholar 

  • Matysik J, Bhalu AB, Mohanty P (2002) Molecular mechanisms of quenching of reactive oxygen species by proline under stress in plants. Curr Sci 82:525–532

    CAS  Google Scholar 

  • Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410

    Article  CAS  PubMed  Google Scholar 

  • Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu Rev Plant Biol 59:651–681

    Article  CAS  PubMed  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nanjo T, Fujita M, Seki M, Kato T, Tabata S, Shinozaki K (2003) Toxicity of free proline revealed in an Arabidopsis T-DNA-tagged mutant deficient in proline dehydrogenase. Plant Cell Physiol 44:541–548

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Noctor G, Gomez L, Vanacker H, Foyer CH (2002) Interactions between biosynthesis, compartmentation and transport in the control of glutathione homeostasis and signalling. J Exp Bot 53:1283–1304

    Article  CAS  PubMed  Google Scholar 

  • Noctor G, Mhamdi A, Chaouch S, Han Y, Neukermans J, Marquez-Garcia B, Queval G, Foyer CH (2012) Glutathione in plants: an integrated overview. Plant, Cell Environ 35:454–484

    Article  CAS  Google Scholar 

  • Nounjan N, Theerakulpisut P (2012) Effects of exogenous proline and trehalose on physiological responses in rice seedlings during salt-stress and after recovery. Plant Soil Environ 58(7):309–315

    CAS  Google Scholar 

  • Ogawa K (2005) Glutathione-associated regulation of plant growth and stress responses. Antioxid Redox Signal 7:973–981

    Article  CAS  PubMed  Google Scholar 

  • Okuma E, Soeda K, Tada M, Murata Y (2000) Exogenous proline mitigates the inhibition of growth of Nicotiana tabacum cultured cells under saline conditions. Soil Sci Plant Nutr 46:257–263

    Article  CAS  Google Scholar 

  • Parida AK, Das AB (2005) Salt tolerance and salinity effects on plants: a review. Ecotox Environ Saf 60:324–349

    Article  CAS  Google Scholar 

  • Parida AK, Das AB, Mohanty P (2004) Investigations on the antioxidative defense responses to stress in a mangrove, Bruguiera parviflora differential regulations of isoforms of some enzymes. Plant Growth Regul 42:213–226

    Article  CAS  Google Scholar 

  • Parida AK, Dagaonkar VS, Phalak MS, Aurangabadkar LP (2008) Differential responses of the enzymes involved in proline biosynthesis and degradation in drought tolerant and sensitive cotton genotypes during drought stress and recovery. Acta Physiol Plant 30:619–662

    Article  CAS  Google Scholar 

  • Pastori GM, Foyer CH (2002) Common components, networks, and pathways of cross-tolerance to stress. The central role of “Redox” and abscisic acid-mediated controls. Plant Physiol 129:460–468

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Pérez-Clemente RM, Gómez-cadenas A (2012) In vitro tissue culture, a tool for the study and breeding of plants subjected to abiotic stress conditions. In Leva A, Rinaldi MRR (eds), Recent advances in plant in vitro culture (pp 92–108). ISBN: 978-953-51-0787-3

  • Rengasamy P (2002) Transient salinity and subsoil constraints to dryland farming in Australian sodic soils: an overview. Aust J Exp Agr 42:351–361

    Article  Google Scholar 

  • Roy D, Basu N, Bhunia A, Banerjee SK (1993) Counteraction of exogenous l-proline with NaCl in salt-sensitive cultivar of rice. Biol Plant 35:69–72

    Article  CAS  Google Scholar 

  • Shao HB, Chu LY, Lu ZH, Kang CM (2008) Primary antioxidant free radical scavenging and redox signalling pathways in higher plant cells. Int J Biol Sci 4:8–14

    Article  PubMed Central  CAS  Google Scholar 

  • Sharma N, Gupta NK, Gupta S, Hasegawa H (2005) Effect of NaCl on photosynthetic rate, transpiration rate and oxidative stress tolerance in contrasting wheat genotypes. Photosynthetic 43:609–613

    Article  Google Scholar 

  • Shibli RA, Al-Juboory K (2002) Comparative responses of ‘Nabali’ olive microshoot, callus and suspension cell culture to salinity and water deflicit. J Plant Nutr 25:61–74

    Article  CAS  Google Scholar 

  • Srivalli S, Khanna-chopra R (2008) Role of glutathione in abiotic stress tolerance. In: Khan NA, Singh S, S Umar (eds) Sulfur assimilation and abiotic stress in plants. Springer, Berlin, pp 207–225

    Chapter  Google Scholar 

  • Subbarao GV, Ito O, Berry WL, Wheeler RM (2003) Sodium- a functional plant nutrient. Crit Rev Plant Sci 22(5):391–416

    Google Scholar 

  • Sun HY, Wang XY, Dai HX, Zhang GP, Wu FB (2012) Effect of exogenous glutathione and selenium on cadmium-induced changes in cadmium and mineral concentrations and antioxidative metabolism in maize seedlings. Asian J Chem 25(6):2970–2976

    Google Scholar 

  • Türkan I, Demiral T (2009) Recent developments in understanding salinity tolerance. Environ Exp Bot 67:2–9

    Article  Google Scholar 

  • Verbruggen N, Hermans C (2008) Proline accumulation in plants: a review. Amino Acids 35(753):759

    Google Scholar 

  • Wang WB, Vinocur Altman A (2003) Plant response to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta 218:1–14

    Article  CAS  PubMed  Google Scholar 

  • Wang RL, Liu SH, Zhou F, Ding CX, Hua C (2014) Exogenous ascorbic acid and glutathione alleviate oxidative stress induced by salt stress in the chloroplast of Oryza sativa L. Z Naturforsch 69c:226–236

    Google Scholar 

  • Yadav SK, Singla-Pareek SL, Reddy MK, Sopory SK (2005) Transgenic tobacco plants overexpressing glyoxalase enzymes resist an increase in methylglyoxal and maintain higher reduced glutathione levels under salinity stress. FEBS Lett 579:6265–6271

    Article  CAS  PubMed  Google Scholar 

  • Yamaguchi T, Blumwald E (2005) Developing salt-tolerant crop plants: challenges and opportunities. Trends Plant Sci 10:615–620

    Article  CAS  PubMed  Google Scholar 

  • Zhu JK (2002) Salt and drought stress signal transduction in plants. Annu Rev Plant Biol 53:247–273

    Article  PubMed Central  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We acknowledge LRGS (Long Term Research Grant Scheme) in Food Security: Enhancing Sustainable Rice Production through Innovative Research for providing the research fund. The author would like to acknowledge MyBrain15 for the scholarship.

Author information

Authors and Affiliations

  1. Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Chui Yao Teh, Maziah Mahmood & Noor Azmi Shaharuddin

  2. Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Chai Ling Ho

  3. Institute of Tropical Agriculture, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Maziah Mahmood

Authors
  1. Chui Yao Teh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Maziah Mahmood
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Noor Azmi Shaharuddin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chai Ling Ho
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Maziah Mahmood.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Teh, C.Y., Mahmood, M., Shaharuddin, N.A. et al. In vitro rice shoot apices as simple model to study the effect of NaCl and the potential of exogenous proline and glutathione in mitigating salinity stress. Plant Growth Regul 75, 771–781 (2015). https://doi.org/10.1007/s10725-014-9980-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10725-014-9980-2

Keywords

Search

Navigation