Acta Physiologiae Plantarum

, 41:23 | Cite as

Proline-mediated changes in antioxidant enzymatic activities and the physiology of sugar beet under drought stress

  • Hamideh Ghaffari
  • Mahmoud Reza TadayonEmail author
  • Muhammad Nadeem
  • Mumtaz Cheema
  • Jamshid Razmjoo
Original Article


Drought stress (DS) is a major concern in the agricultural sector and, in particular, for sugar beet production and sugar content. As such several agricultural practices have been used to minimize yield losses from DS, and foliar application of proline is considered one such approach to improve drought tolerance in growing plants. Hence, the current study examined the proline-related improvements to induce drought tolerance in sugar beet plants. A field experiment was conducted at two locations (Shahrekord and Shalamzar) in Chaharmahal-Bakhtiari province, Iran. Experimental treatments comprised of three DS levels (well water: 100%; mild stress: 75%; severe stress: 50% water requirement of plant), and three proline applications (control: 0; low: 5 mM; high: 10 mM). DS caused a significant up-regulation in leaf proline content, malondialdehyde (MDA) content, hydrogen peroxide (H2O2) content, ascorbate peroxidase, catalase, and peroxidase enzymatic activities. This increase was more pronounced under proline application with concomitant down-regulation of MDA and H2O2 contents. DS also caused a decrease in leaf photosynthetic pigments, leaf relative water contents, membrane stability index and sugar beet root production; however, proline application mitigated these adverse DS effects. The study results suggest beneficial effects of proline applications, which is crucial to mitigation of the detrimental effects of DS in sugar beet by enhancing antioxidant enzymatic activities with concomitant reduction in MDA and H2O2 contents.


Leaf photosynthetic pigments Enzymes activities Drought stress Root yield 



Ascorbate peroxidase






Drought stress


Hydrogen peroxide




Membrane stability index




Relative water contents


Drought stress




Mild stress


Severe stress



  1. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126PubMedCrossRefGoogle Scholar
  2. Ahmed CB, Rouina BB, Sensoy S, Boukhrissm Abdullah FB (2010) Exogenous proline effects on photosynthetic performance and antioxidant defense system of young olive trees. J Agric Food Chem 58:4216–4222PubMedCrossRefGoogle Scholar
  3. Ali Q, Ashraf M, Athar HUR (2007) Exogenously applied proline at different growth stages enhances growth of two maize cultivars grown under water deficit conditions. Pak J Bot 39(4):1133–1144Google Scholar
  4. AL-Jbawi E, Abbas F (2013) The effect of length during water stress on sugar beet (Beta vulgaris L.) yield and quality. Persian Gulf Crop Protect 2(1):35–43Google Scholar
  5. Allen RG, Pereira LS, Raes D, Smith M (2000) FAO irrigation and drainage paper. Crop evapotranspiration 56:1–326Google Scholar
  6. Al-Shaheen MR, Soh A (2016) Effect of proline and gibberellic acid on the qualities and qualitative of corn (Zea maize L.) under the influence of different levels of the water stress. Int J Sci Res 6(5):752–756Google Scholar
  7. Arnon I (1996) Crop production in dry regions. Leonard Hill, London, 650 ppGoogle Scholar
  8. Ashraf M, Foolad MR (2007) Roles of glycinebetaine and proline in improving plant abiotic stress tolerance. Environ Exp Bot 59:206–216CrossRefGoogle Scholar
  9. Bajji M, Lutts S, Kinet JM (2000) Physiological changes after exposure to and recovery from polyethylene glycol-induced water deficit in callus cultures issued from durum wheat (Triticum durum Desf.) cultivars differing in drought resistance. J Plant Physiol 156:75–83CrossRefGoogle Scholar
  10. Baker NR, Harbinson J, Kramer DM (2007) Determining the limitations and regulation of photosynthetic energy transduction in leaves. Plant Cell Environ 30:1107–1125PubMedCrossRefGoogle Scholar
  11. Banchio E, Bogino PC, Zygadlo J, Giordano W (2008) Plant growth promoting rhizobacteria improves growth and essential oil yield in Origanum majorana L. Biochem Syst Ecol 36:766–771CrossRefGoogle Scholar
  12. Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39(1):205–207CrossRefGoogle Scholar
  13. Ben AC, Ben Rouina B, Sensoy S, Boukhriss M, Ben Abdullah F (2010) Exogenous proline effects on photosynthetic performance and antioxidant defense system of young olive tree. J Agric Food Chem 58:4216–4222CrossRefGoogle Scholar
  14. Boaretto LF, Carvalho G, Borgo L, Creste S, Landell MG, Mazzafera P, Azevedo RA (2014) “Water stress reveals differential antioxidant responses of tolerant and non-tolerant sugarcane genotypes. Plant Physiol Biochem 74:165–175PubMedCrossRefGoogle Scholar
  15. 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 72:248–254CrossRefGoogle Scholar
  16. Chen CT, Chen L, Lin CC, Kao CH (2001) Regulation of proline accumulation in detached rice leaves exposed to excess copper. Plant Sci 160:283–290PubMedCrossRefGoogle Scholar
  17. Chrominski A, Halls S, Weber DJ, Smith BN (1989) Proline affects ACC to ethylene conversion under salt and water stresses in the halophyte Allenrolfea occidentalis. Environ Exp Bot 29:359–363CrossRefGoogle Scholar
  18. Dadkhah AR, Grrifiths H (2006) The effect of salinity on growth, inorganic ions and dry matter partitioning in sugar beet cultivars. J Agric Sci Tech 8:199–210Google Scholar
  19. Dawood MG (2016) Influence of osmoregulators on plant tolerance to water stress. Sci Agric 13(1):42–58Google Scholar
  20. Delauney AJ, Verma DPS (1993) Proline biosynthesis and osmoregulation in plants. Plant J 4:215–223CrossRefGoogle Scholar
  21. Farissi M, Bouizgaren A, Faghire M, Bargaz A, Ghoulam C (2013) Agrophysiological and biochemical properties associated with adaptation of Medicago sativa populations to water deficit. Turk J Bot 37:1166–1175CrossRefGoogle Scholar
  22. Foroozesh P, Majidi Heravan E, Bihamta MR, Fatollah Taleghani D, Habibi D (2012) Physiological evaluation of sugar beet genotypes under water stress. Am Eurasian J Agric Environ Sci 12:820–826Google Scholar
  23. Handa S, Handa AK, Hasegawa PM, Bressan RA (1986) Proline accumulation and the adaptation of cultured plant cells to water stress. Plant Physiol 80:938–945PubMedPubMedCentralCrossRefGoogle Scholar
  24. Hare PD. Cress WA (1997) Metabolic implications of stress induced proline accumulation in plants. Plants Growth Regul 21:79–102CrossRefGoogle Scholar
  25. Hare PD, Cress WA, Van Staden J (2003) A regulatory role for proline metabolism in stimulating Arabidopsis thaliana seed germination. Plant Growth Regul 39:41–50CrossRefGoogle Scholar
  26. Hasanuzzaman M, Alam MM, Rahman A, Hasanuzzaman M, Nahar K, Fujita M (2014) Exogenous proline and glycine betaine mediated upregulation of antioxidant defense and glyoxalase systems provides better protection against salt-induced oxidative stress in two rice (Oryza sativa L.) varieties. BioMed Res Int 757219:1–17Google Scholar
  27. Hayat S, Hayat Q, Alyemeni MN, Wani AS, Pichtel J, Ahmad A (2012) Role of proline under changing environments. A review. Plant Signal Behav 7:1456–1466PubMedPubMedCentralCrossRefGoogle Scholar
  28. Herzog V, Fahimi H (1973) Determination of the activity of peroxidase. Anal Biochem 55:554–562PubMedCrossRefGoogle Scholar
  29. Hong Z, Lakkineni K, Zhang Z, Verma DP (2000) Removal of feedback inhibition of delta(1)-pyrroline-5-carboxylate synthetase results in increased proline accumulation and protection of plants from osmotic stress. Plant Physiol 122:1129–1136PubMedPubMedCentralCrossRefGoogle Scholar
  30. Hoque MA, Banu MN, Okuma E, Amako K, Nakamura Y, Shimoishi Y, Murata Y (2007) Exogenous proline and glycinebetaine increase NaCl-induced ascorbate–glutathione cycle enzyme activities, and proline improves salt tolerance more than glycinebetaine in tobacco Bright Yellow-2 suspension-cultured cells. J Plant Physiol 164:1457–1468PubMedCrossRefGoogle Scholar
  31. Hossain MA, Fujita M (2010) Evidence for a role of exogenous glycinebetaine and proline in antioxidant defense and methylglyoxal detoxification systems in mung bean seedlings under salt stress. Physiol Mol Biol Plants 16(1):19–29PubMedPubMedCentralCrossRefGoogle Scholar
  32. Imlay JA (2002) Cellular defense against superoxide and hydrogen peroxide. Annl Rev Biochem 77:755–776CrossRefGoogle Scholar
  33. Irigoyen JJ, Emerich DW, Sanchez-Diaz M (1992) Water stress induce changes in concentration of proline and total soluble sugar in nodulated alfalfa (Medicago sativa) plants. Physiol Plant 84:55–60CrossRefGoogle Scholar
  34. Irshad M, Honna T, Eneji AE, Yamamoto S (2002) Wheat response to nitrogen source under saline conditions. J Plant Nutr 2512:2603–2612CrossRefGoogle Scholar
  35. Islam MM, Hoque MA, Okuma E, Banu MN, Shimoishi Y, Nakamura Y, Murata Y (2009) Exogenous proline and glycinebetaine increase antioxidant enzyme activities and confer tolerance to cadmium stress in cultured tobacco cells. J Plant Physiol 166:1587–1597PubMedCrossRefGoogle Scholar
  36. Jinyou D, Xiaoyang C, Wei L, Qiong G (2004) Osmoregulation mechanism of drought stress and genetic engineering strategies for improving drought resistance in plants. For Stud China 6(2):56–62CrossRefGoogle Scholar
  37. Kadkhodaie A, Zahedi M, Razmjoo J, Pessarakli M (2014) Changes in some anti-oxidative enzymes and physiological indices among sesame genotypes (Sesamum indicum L.) in response to soil water deficits under field conditions. Acta Physiol Plant 36:641–650CrossRefGoogle Scholar
  38. Kibria MG, Farzana K, Abdul Matin M, Anamul Hoque MA (2016) Mitigating water stress in wheat (BARI Gom-26) by exogenous application of proline. Fundam Appl Agric 1(3):118–123Google Scholar
  39. Kishor PBK, Sangam S, Amrutha RN, Sri Laxmi P, Naidu KR, Rao KRSS, Rao S, 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. Review Curr Sci 88(3):424–438Google Scholar
  40. Kumar NS, Zhu W, Liang X, Zhang L, Demers AJ, Zimmerman MC, Simpson MA, Becker DF (2012) Proline dehydrogenase is essential for proline protection against hydrogen peroxide-induced cell death. Free Radic Biol Med 53:1181–1191CrossRefGoogle Scholar
  41. Liang X, Zhang L, Natarajan SK, Becker D (2013) Proline mechanisms of stress survival. Antioxid Redox Signal 19(9):998–1011PubMedPubMedCentralCrossRefGoogle Scholar
  42. Liu C, Liu Y, Guo K, Fan D, Li G, Zheng Y, Yu L, Yang R (2011) Effect of drought on pigments, osmotic adjustment and antioxidant enzymes in six woody plant species in karst habitats of southwestern China. Environ Exp Bot 71:174–183CrossRefGoogle Scholar
  43. Madhava KV, Sresty TVS (2000) Antioxidative parameters in the seedlings of pigeonpea (Cajanus cajan (L.) Millspaugh) in response to Zn and Ni stresses. Plant Sci 157:113–128CrossRefGoogle Scholar
  44. Mafakheri A, Siosemardeh A, Bahramnejad B, Struik PC, Sohrabi Y (2010) Effect of drought stress on yield, proline and chlorophyll contents in three chickpea cultivars. Aus J Crop Sci 4(8):580–585Google Scholar
  45. Mansour MMF (2000) Nitrogen containing compound and adaptation of plants to salinity stress. Biol Plant 43:491–500CrossRefGoogle Scholar
  46. Mansuri M, Babazadeh H, Emdad MR, Taleghani D (2017) effect of deficit irrigation management on qualitative and quantitative yield of sugar beet (Beta vulgaris L.) in Karaj, Iran. Appl Ecol Env Res 16(1):455–466CrossRefGoogle Scholar
  47. Marcińska I, Czyczyło-Mysza I, Skrzypek E, Grzesiak MT, Janowiak F, Filek M, Dziurka M, Dziurka K, Waligórski P (2013) Alleviation of osmotic drought effects by exogenous application of salicylic or abscisic acid on wheat seedlings. Int J Mol Sci 14:13171–13193PubMedPubMedCentralCrossRefGoogle Scholar
  48. Matysik J, Alia BB, Monthy P (2002) Molecular mechanisms of quenching of reactive oxygen species by proline under stress in plants. Curr Sci 82:525–532Google Scholar
  49. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22:867–880Google Scholar
  50. Ober ES, Rajabi A (2010) Abiotic stress in sugar beet. Sugar Tech 12:294–298CrossRefGoogle Scholar
  51. Ozturk L, Demir Y (2002) In vivo and in vitro protective role of proline. Plant Growth Regul 38:259–264CrossRefGoogle Scholar
  52. Phang JM, Liu W, Zabirnyk O (2010) Proline metabolism and microenvironmental stress. Annu Rev Nutr 30:441–463PubMedCrossRefGoogle Scholar
  53. Raymond MJ, Smirnoff N (2002) Proline metabolism and transport in maize seedlings at low water potential. Annals Bot 89:813–823CrossRefGoogle Scholar
  54. Refay YA (2010) Root yield and quality traits of three sugar beet (Beta vulgaris L.) varieties in relation to sowing date and stand densities. World J Agri Sci 6(5):589–594Google Scholar
  55. Reza S, Heidari R, Zare S, Norastehnia A (2006) Antioxidant response of two salt-stressed barley varieties in the presence or absence of exogenous proline. Gen App Plant Physiol 32:233–251Google Scholar
  56. Rontein D, Basset G, Hanson AD (2002) Metabolic engineering of osmoprotectant accumulation in plants. Metab Eng 4:49–56PubMedCrossRefGoogle Scholar
  57. SAS Institute (2002) The SAS system for windows. In: Released 9.1. SAS Inst., Cary, NCGoogle Scholar
  58. Sims DA, Gamon JA (2002) Relationships between leaf pigment content and spectral reflectance across a wide range of species, leaf structures and developmental stages. Remote Sens Environ 81:337–354CrossRefGoogle Scholar
  59. Srivastava AK, Suprasanna P, Srivastava S, D’Souza SF (2010) Thiourea mediated regulation in the expression profile of aquaporins and its impact on water homeostasis under salinity stress in Brassica juncea roots. Plant Sci 178:517–522CrossRefGoogle Scholar
  60. Valentovic P, Luxova M, Kolarovic L, Gasparikova O (2006) Effect of osmotic stress on compatible solutes content, membrane stability and water relations in two maize cultivars. Plant Soil Environ 52(4):186–191CrossRefGoogle Scholar
  61. Verbruggen N, Hermans C (2008) Proline accumulation in plants: a review. Amino Acids 35:753–759PubMedCrossRefGoogle Scholar
  62. Yang SL, Lan SS, Gong M (2009) Hydrogen peroxide-induced proline and metabolic pathway of its accumulation in maize seedlings. J Plant Physiol 166:1694–1699PubMedCrossRefGoogle Scholar
  63. Yang Y, Zhang Y, Wei X, You J, Wang W, Lu J, Shi R (2011) Comparative antioxidative responses and proline metabolism in two wheat cultivars under short term lead stress. Ecotoxicol Environ Saf 74:733–740PubMedCrossRefGoogle Scholar
  64. Yeilaghi H, Arzani A, Ghaderian M, Fotovat R, Feizi M, Pourdad SS (2012) Effect of salinity on seed oil content and fatty acid composition of safflower (Carthamus tinctorius L.) genotypes. Food Chem 130:618–625CrossRefGoogle Scholar
  65. Yordanov I, Velikova V, Tsonev T (2000) Plant responses to drought, acclimation, and stress tolerance. Photosynthetica 38:171–186CrossRefGoogle Scholar
  66. Yu CW, Murphy TM, Lin CH (2003) Hydrogen peroxide induces chilling tolerance in mung beans mediated through ABA-independent glutathione accumulation. Funct Plant Biol 30:955–963CrossRefGoogle Scholar
  67. Zali AG, Ehsanzadeh P (2018) Exogenous proline improves osmoregulation, physiological functions, essential oil, and seed yield of fennel. Ind Crops Prod 111:133–140CrossRefGoogle Scholar

Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2019

Authors and Affiliations

  • Hamideh Ghaffari
    • 1
  • Mahmoud Reza Tadayon
    • 1
    Email author
  • Muhammad Nadeem
    • 2
    • 4
  • Mumtaz Cheema
    • 2
  • Jamshid Razmjoo
    • 3
  1. 1.Department of Agronomy, Faculty of AgricultureShahrekord UniversityShahrekordIran
  2. 2.School of Science and the Environment, Grenfell CampusMemorial University of NewfoundlandCorner BrookCanada
  3. 3.Department of Agronomy and Plant Breeding, Faculty of AgricultureIsfahan University of TechnologyIsfahanIran
  4. 4.Department of Environmental SciencesCOMSATS University of IslamabadIslamabadPakistan

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