Acta Physiologiae Plantarum

, 33:2091 | Cite as

Heat-stress induced inhibition in growth and chlorosis in mungbean (Phaseolus aureus Roxb.) is partly mitigated by ascorbic acid application and is related to reduction in oxidative stress

  • Sanjeev Kumar
  • Ramanpreet Kaur
  • Navneet Kaur
  • Kalpna Bhandhari
  • Neeru Kaushal
  • Kriti Gupta
  • T. S. Bains
  • Harsh Nayyar
Original Paper


The rising temperatures (>35°C) are proving detrimental to summer-sown mungbean genotypes that experience inhibition of vegetative and reproductive growth. In the present study, the mungbean plants growing hydroponically at varying temperatures of 30/20°C (control), 35/25, 40/30, and 45/35°C (as day/night 12 h/12 h) with (50 μM) or without ascorbic acid (ASC) were investigated for effects on growth, membrane damage, chlorophyll loss, leaf water status, components of oxidative stress, and antioxidants. The ASC-treated plants showed significant improvement in germination and seedling growth especially at 40/30 and 45/35°C. The damage to membranes, loss of water, decrease in cellular respiration, and chlorophyll were significantly prevented by ASC treatment to plants growing at these temperatures. The oxidative stress measured as malondialdehyde and hydrogen peroxide content was observed to be significantly lower at high temperatures with ASC application. The activities of superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase increased at 40/30°C but decreased at 45/35°C in the absence of ASC while with its application, the activities of these enzymes were appreciably resorted. Among all the antioxidants, the endogenous ASC content decreased to the greatest extent at 45/35°C grown plants indicating its vital role in affecting the response of mungbean to heat stress. Exogenously applied ASC raised its endogenous content along with that of glutathione and proline at 45/35°C. The findings indicated that heat stress-induced inhibition in growth and chlorosis was associated with decrease in leaf water status and elevation of oxidative stress, which could partly be prevented by exogenous application of ASC. Its role in imparting protection against heat stress is discussed.


Ascorbic acid High temperature Mungbean Oxidative stress 


  1. Almeselmani M, Deshmukh PS, Sairam RK (2009) High temperature stress tolerance in wheat genotypes: role of antioxidant defence enzymes. Acta Agron Hung 57:1–14CrossRefGoogle Scholar
  2. Arnon DI (1949) Copper enzyme in isolated chloroplasts: polyphenol oxidase in Beta vulgaris. Plant Physiol 24:1–15PubMedCrossRefGoogle Scholar
  3. Balla K, Bedő Z, Veisz O (2007) Heat stress induced changes in the activity of antioxidant enzymes in wheat. Cereal Res Comm 35:197–200CrossRefGoogle Scholar
  4. Barrs HD, Weatherley PE (1962) A re-examination of the relative turgidity techniques for estimating water deficits in leaves. Aust J Biol Sci 15:413–428Google Scholar
  5. Basra RK, Basra AS, Malik CP, Grover IS (1997) Are polyamines involved in the heat-shock protection of mung bean seedlings? Bot Bull Acad Sin 38:165–169Google Scholar
  6. Bates LS, Woldren RP, Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205–208CrossRefGoogle Scholar
  7. Camejo D, Torres W (2001) High temperature effect on tomato (Lycopersicon esculentum) pigment and protein content and cellular viability. Cultivos Trop 22:13–17Google Scholar
  8. Cao YY, Duan H, Yang L, Wang Z, Zhou SC, Yang JC (2008) Effect of heat-stress during meiosis on grain yield of rice cultivars differing in heat-tolerance and its physiological mechanism. Acta Agron Sin 34:2134–2142CrossRefGoogle Scholar
  9. Chaitanya V, Sundar D, Reddy AR (2001) Mulberry leaf metabolism under high temperature stress. Biol Plant 44:379–384CrossRefGoogle Scholar
  10. Chang YC, Lee TM (1999) High temperature-induced free proline accumulation in Gracilaria tenuistipitata (Rhodophyta). Bot Bull Acad Sin 40:289–294Google Scholar
  11. Change B, Maehly AC (1955) Assay of catalases and peroxidase. Methods Enzymol 2:764–775CrossRefGoogle Scholar
  12. Chen HH, Shen ZY, Li PH (1982) Adaptability of crop plants to high temperature stress. Crop Sci 22:719–725CrossRefGoogle Scholar
  13. Coria NA, Sarquís JI, Peñalosa I, Urzúa M (1998) Heat-induced damage in potato (Solanum tuberosum) tubers: membrane stability, tissue viability, and accumulation of glycoalkaloids. J Agric Food Chem 46:4524–4528Google Scholar
  14. Dash S, Mohanty N (2002) Response of seedlings to heat-stress in cultivars of wheat: Growth temperature-dependent differential modulation of photosystem 1 and 2 activity, and foliar antioxidant defense capacity. J Plant Physiol 159:49–59CrossRefGoogle Scholar
  15. Dionisio-Sese ML, Shono M, Tobita S (1999) Effects of proline and betaine on heat inactivation of ribulose-1,5-bisphosphate carboxylase/oxygenase in crude extracts of rice seedlings. Photosynthetica 36:557–563CrossRefGoogle Scholar
  16. Dolatabadian A, Sanavy SAMM, Chashmi NA (2008) The effects of foliar application of ascorbic acid (Vitamin C) on antioxidant enzymes activities lipid peroxidation and proline accumulation of Canola (Brassica napus L.) under conditions of salt stress. J Agron Crop Sci. 194:206–213CrossRefGoogle Scholar
  17. Dolatabadian A, Sanavy SAMM, Sharifi M (2009) Alleviation of water deficit stress effects by foliar application of ascorbic acid on Zea mays L. J Agron Crop Sci 195:347–355CrossRefGoogle Scholar
  18. Foyer CH, Halliwell B (1976) The presence of glutathione and glutathione reductase in chloroplasts: a proposed role in ascorbic acid metabolism. Planta 133:21–25CrossRefGoogle Scholar
  19. Giannopolities CN, Ries SK (1977) Superoxide dismutase. I. Occurrence in higher plants. Plant Physiol 59:309–314CrossRefGoogle Scholar
  20. Giaveno C, Ferrero J (2003) Introduction of tropical maize genotypes to increase silage production in the central area of Santa Fe, Argentina. Crop Breed Appl Biotechnol 3:89–94Google Scholar
  21. Griffth OW (1980) Determination of glutathione and glutathione disulfide using glutathione reductase and 2 vinyl pyridine. Anal Biochem 106:207–212CrossRefGoogle Scholar
  22. Gulen H, Eris A (2004) Effect of heat stress on peroxidase activity and total protein content in strawberry plants. Plant Sci 166:739–744CrossRefGoogle Scholar
  23. Guo YP, Zhou HF, Zhang LC (2006) Photosynthetic characteristics and protective mechanisms against photooxidation during high temperature stress in two citrus species. Sci Hortic 108:260–267CrossRefGoogle Scholar
  24. Gür A, Demirel U, Özden M, Kahraman A, Çopur O (2010) Diurnal gradual heat stress affects antioxidant enzymes, proline accumulation and some physiological components in cotton (Gossypium hirsutum L.). Afr J Biotechnol 9:1008–1015Google Scholar
  25. Hall AE (2001) Breeding for heat tolerance. Plant Breed Rev 10:129–168Google Scholar
  26. Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplast. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198PubMedCrossRefGoogle Scholar
  27. Howarth CJ (2005) Genetic improvements of tolerance to high temperatures. In: Ashraf M, Harris PJC (eds) Abiotic stresses: plant resistance through Breeding and molecular approaches. Howarth Press Inc., New YorkGoogle Scholar
  28. Ibrahim MHA, Quick JS (2001) Heritability of heat tolerance in winter and spring wheat. Crop Sci 41:1401–1405CrossRefGoogle Scholar
  29. Jiang Y, Huang B (2001) Drought and heat stress injury to two cool-season turfgrasses in relation to antioxidant metabolism and lipid peroxidation. Crop Sci 41:436–442CrossRefGoogle Scholar
  30. Kepova KD, Hozler R, Stoilova LS, Feller U (2005) Heat stress effects on ribulose-1,5,bisphosphate carboxylase/oxygenase, Rubisco binding protein and Rubisco activase in wheat leaves. Biol Plant 49:521–525CrossRefGoogle Scholar
  31. Kumar RA, Deshmukh TP, Singh SR, Kushwaha SR, Bhagat K (2008) Superoxide dismutase in chickpea genotypes under high temperature. Ind J Plant Physiol 13:88–90Google Scholar
  32. Lechno S, Zamski E, Tel-Or E (1997) Salt stress-induced responses in cucumber plants. J Plant Physiol 150:206–211Google Scholar
  33. Liu X, Huang B (2000) Heat stress injury in relation to membrane lipid peroxidation in creeping bentgrass. Crop Sci 40:503–510CrossRefGoogle Scholar
  34. Liu J, Xie J, Du J, Sun J, Bai X (2008) Effects of simultaneous drought and heat stress on Kentucky bluegrass. Sci Hortic 115:190–195CrossRefGoogle Scholar
  35. Ma YH, Ma FW, Zhang JK, Li MJ, Wang YH, Liang D (2008) Effects of high temperature on activities and gene expression of enzymes involved in ascorbate–glutathione cycle in apple leaves. Plant Sci 175:761–766CrossRefGoogle Scholar
  36. Mahan JR, Mauget SA (2005) Antioxidant metabolism in cotton seedlings exposed to temperature stress in the field. Crop Sci 45:2337–2345CrossRefGoogle Scholar
  37. Mishra S, Heckathorn SA, Barua D, Wang D, Joshi P, Hamilton EW III, Frantz J (2008) Interactive effects of elevated CO2 and ozone on leaf thermotolerance in field-grown Glycine max (plants and global environmental change: a special issue highlighting younger scientists.). J Integr Plant Biol 50:1396–1405PubMedCrossRefGoogle Scholar
  38. Morales D, Rodriguez P, Dell’amico J, Nicolas E, Torrecillas A, Sanchez-Blanco MJ (2003) High temperature pre-conditioning and thermal shock imposition affects water relations, gas exchange and root hydraulic conductivity in tomato. Biol Plant 47:203–208CrossRefGoogle Scholar
  39. Mukherjee SP, Choudhuri MA (1983) Implications of water stress induced changes in the levels of endogenous ascorbic acid and hydrogen peroxide in Vigna seedlings. Physiol Plant 58:166–170CrossRefGoogle Scholar
  40. Nagesh Babu R, Devaraj VR (2008) High temperature and salt stress response in french bean (Phaseolus vulgaris). Aust J Crop Sci 2:40–48Google Scholar
  41. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22:867–880Google Scholar
  42. Nayyar H, Gupta D (2006) Differential sensitivity of C3 and C4 plants to water deficit stress: association with oxidative stress and antioxidants. Environ Exp Bot 58:106–113CrossRefGoogle Scholar
  43. Nayyar H, Kaushal SK (2002) Chilling induced oxidative stress in germinating wheat grains as affected by water stress and calcium. Biol Plant 45:601–604CrossRefGoogle Scholar
  44. Ortiz C, Cardemil L (2001) Heat-shock responses in two leguminous plants: a comparative study. J Exp Bot 52:1711–1719PubMedCrossRefGoogle Scholar
  45. Premchandra GS, Sameoka H, Ogata S (1990) Cell osmotic membrane-stability, an indication of drought tolerance, as affected by applied nitrogen in soil. J Agric Res 115:63–66Google Scholar
  46. Rivero RM, Ruiz JM, Garcia PC, Lopez-Lefebre LR, Sanchez E, Romero L (2001) Resistance to cold and heat stress: accumulation of phenolic compounds in tomato and water melon plants. Plant Sci 160:315–321PubMedCrossRefGoogle Scholar
  47. Sairam RK, Srivastava GC, Saxena DC (2000) Increased antioxidant activity under elevated temperatures: a mechanism of heat stress tolerance in wheat genotypes. Biol Plant 43:245–251CrossRefGoogle Scholar
  48. Shalata A, Neumann PM (2001) Exogenous ascorbic acid (vitamin C) increases resistance to salt stress and reduces lipid peroxidation. J Exp Bot 52:2207–2211PubMedGoogle Scholar
  49. Singh DP, Ahlawat IPS (2005) Greengram and blackgram improvement in India: past, present and future prospects. Indian J Agric Sci 75:243–250Google Scholar
  50. Smirnoff N (1996) The function and metabolism of ascorbic acid in plants. Ann Bot 78:661–669CrossRefGoogle Scholar
  51. Sohn SO, Back K (2007) Transgenic rice tolerant to high temperature with elevated contents of dienoic fatty acids. Biol Plant 51:340–342CrossRefGoogle Scholar
  52. Stasolla C, Yeung EC (1999) Ascorbic acid improves conversion of white spruce somatic embryos. In Vitro Cell Dev Biol Plant 35:316–319CrossRefGoogle Scholar
  53. Steponkus PL, Lanphear FO (1967) Refinement of the triphenyl tetrazolium chloride method of determining cold injury. Plant Physiol 42:1423–1426PubMedCrossRefGoogle Scholar
  54. Sung DY, Kaplan F, Lee KJ, Guy CL (2003) Acquired tolerance to temperature extremes. Trends Plant Sci 8:179–187PubMedCrossRefGoogle Scholar
  55. Thind SK, Chanpreet, Mridula (1997) Effect of fluridone on free sugar level in heat stressed mungbean seedlings. Plant Growth Regul 22:19–22CrossRefGoogle Scholar
  56. Upadhyaya HCP, Akula N, Young KE, Chun SC, Kim DH, Park SW (2010) Enhanced ascorbic acid accumulation in transgenic potato confers tolerance to various abiotic stresses. Biotechnol Lett 32:321–330PubMedCrossRefGoogle Scholar
  57. Vollenweider P, Gunhardt-Goerg MS (2005) Diagnosis of abiotic and biotic stress factors using the visible symptoms in foliage. Environ Pollut 137:455–465PubMedCrossRefGoogle Scholar
  58. Wahid A, Gelani S, Ashraf M, Foolad MR (2007) Heat tolerance in plants: an overview. Environ Exp Bot 61:199–223CrossRefGoogle Scholar
  59. Wang WC, Nguyen HT (1989) Thermal stress evaluation of suspension cell cultures in winter wheat. Plant Cell Rep 8:108–111CrossRefGoogle Scholar
  60. Wang SY, Zheng W (2001) Effect of plant growth temperature on antioxidant capacity in strawberry. J Agric Food Chem 49:4977–4982PubMedCrossRefGoogle Scholar
  61. Yin H, Chen Q, Yi M (2008) Effects of short-term heat stress on oxidative damage and responses of antioxidant system in Lilium longiflorum. Plant Growth Regul 54:45–54CrossRefGoogle Scholar
  62. Younis ME, Hasaneen MNA, Kazamel AMS (2010) Exogenously applied ascorbic acid ameliorates detrimental effects of NaCl and mannitol stress in Vicia faba seedlings. Protoplasma 239:39–48PubMedCrossRefGoogle Scholar
  63. Zhang J, Kirkham MB (1996) Sorghum and sunflower seedlings as affected by ascorbic acid, benzoic acid and propyl gallate. J Plant Physiol 149:489–493Google Scholar

Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2011

Authors and Affiliations

  • Sanjeev Kumar
    • 1
  • Ramanpreet Kaur
    • 1
  • Navneet Kaur
    • 1
  • Kalpna Bhandhari
    • 1
  • Neeru Kaushal
    • 1
  • Kriti Gupta
    • 1
  • T. S. Bains
    • 2
  • Harsh Nayyar
    • 1
  1. 1.Department of BotanyPanjab UniversityChandigarhIndia
  2. 2.Department of Plant BreedingP.A.U.LudhianaIndia

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