Water, Air, & Soil Pollution

, 228:376 | Cite as

Physiological Characteristics of Medicago sativa L. in Response to Acid Deposition and Freeze-Thaw Stress

  • Guozhang BaoEmail author
  • Qi Ao
  • Qiqi Li
  • Yishu Bao
  • Yue Zheng
  • Xiaoxia Feng
  • Xuemei Ding


Acid deposition and temperature variation could lead to changes of physiological characteristics of plants in response to stress. In this paper, Medicago sativa CV. Dongmu–1 was investigated to test the effects of freeze-thaw circle and acid deposition upon the changes of osmotic adjustment substances, biological membrane permeability, and antioxidant enzymes. The experiment was conducted under laboratory conditions, and the seedlings were divided into four groups (group I: no treatment, group II: acid stressed only, group III: freeze-thaw stressed only, group IV: both freeze-thaw and acid stressed). Results indicated that under freeze-thaw circle and acid deposition, the contents of malondialdehyde (MDA) and proline increased respectively by 0.6~203.4 and 19.3~68.8% when compared with group I, while protein content declined by 4.1~31.7%, and the effects were even significant than freeze-thaw-only stressed groups. In the freeze-thaw process, superoxide dismutase (SOD) activity dropped at first and then increased with the increase of temperature, peaking at − 3 °C by 1118.45 U g−1; peroxidase (POD) activity showed a brief rise and declined rapidly below 0 °C. By increasing the potentials of antioxidant enzymes and MDA, the membrane lipid peroxidation inside alfalfa was prevented; meanwhile, several indexes changed adaptively in resisting hurts. Variation of SOD and POD was induced by the defense mechanism, which showed alfalfa’s satisfactory cold resistance and acid tolerance. Further research on acid deposition and freeze-thaw circle would be beneficial for the global cultivation of forage grass.


Medicago sativa LFreeze-thaw Acid deposition Physical characteristics 



This work was sponsored by the National Natural Science Foundation of China (Grant Nos. 31772669 and 31270367) and Natural Science Foundation of Jilin Province of China (Grant No. 20150101089JC).


  1. Adem, G., Metin, T., Nurgul, K., Sefik Tufenkci, M., Kerim, M. C., Ertan, Y., & Sezai, E. (2016). Effects of bio-bor fertilizer applications on fruit yield, antioxidant enzyme activity and freeze injury of strawberry. Erwerbs Obstbau, 58, 177.CrossRefGoogle Scholar
  2. Chi, C., Ding, G. H., & Lian, Y. Q. (2007). Effect of low temperature stress on proline content and membrane permeability in three kind cold-season turfgrass. Chinese Agricultural Science Bulletin, 23(01), 101–104.Google Scholar
  3. Fan, H. B. (2002). On worldwide acid rain research. Journal of Fujian College of Forestry, 22(4), 371–375.Google Scholar
  4. Feng, Z. W. (2000). Impacts and control strategies of acid deposition on terrestrial ecosystems in China. Engineering Science, 2(9), 5–11.Google Scholar
  5. Feng, C. J., Luo, X. Y., & Sha, W. (2005). Effect of low temperature stress on SOD, POD activity and proline content of alfalfa. Pratacultural Science, 22(6), 29–32.Google Scholar
  6. Fleck, R. A., Day, J. G., & Clarke, K. J. (1990). Elucidation of the metabolic and structural basis for the cryopreservation recalcitrance of Vaucheria sessilis. CryoLetters, 20, 271–282.Google Scholar
  7. Gao, Y., Qi, X. H., Yang, J. H., & Zheng, M. F. (2007). The response mechanism of cold stress in higher plants. Northern Horticulture, 10, 58–61.Google Scholar
  8. Guy, C. L. (2003). Freezing tolerance of plants: current understanding and selected emerging concepts. Canadian Journal of Botany, 81, 1216–1223.CrossRefGoogle Scholar
  9. Jiang, X. M., Qin, Y., & Guo, G. Z. (2013). Effects of exogenous substances on seed germinationand chilling resistance of pepper under low temperature stress. Xinjiang Agricultural University, 50(12), 2266–2273. Google Scholar
  10. Jing, Y., Jing, C., Li, Y. P., Huang, R., Lu, Q. W., Wang, X. T., Liu, L. J., Xu, Q. H., & Zhang, K. J. (2016). Salicylic acid-induced antioxidant protection against low temperature in cold-hardy winter wheat. Acta Physiologiae Plantarum, 38, 261.CrossRefGoogle Scholar
  11. Li, H. (1988). The state of acid rain in China. World Environment, (04):14–15. Google Scholar
  12. Li, J. H., Yu, Q., & Dou, Z. J. (1985). The effect of free froline treated under low temperature during reproductive period of rice varieties. Journal of Central China Normal University, 3, 1–84. Google Scholar
  13. Li, D., Tang, L., & Zhou, Q. (2008). Effect of the simulated acid rain on the soluble protein and proline contents of rice seeds during the germination. Journal of Safety and Environment, 08(5), 12–15.Google Scholar
  14. Li, S. L., Jia, X. Y., & Li, Y. P. (2009). Research on the characteristics of leaves and cold tolerance of alfalfa by treating with Ca(NO3)2. Journal of Heilongjiang Bayi Agricultural University, 21(3), 9–11.Google Scholar
  15. Liang, J., Mai, B. R., & Zheng, Y. F. (2008). Effects of simulated acid rain on the growth yield and quality of rape. Acta Ecologica Sinica, 28(1), 274–283.Google Scholar
  16. Lin, L. D. (2008). Research on resistance to coldness of lawn grasses of cold season in chilly zones of the northeast. Journal of Qiqihar University, 24(3), 83–85.Google Scholar
  17. Lin, S. Z., Li, X. P., & Zhang, Z. Y. (2002). The effects of cold acclimation on the freezing resistance and total soluble protein in Populus tomentosa seedlings. Scientia Silvae Sinicae, 18(6), 137–141.Google Scholar
  18. Liu, X. H., & Hao, M. D. (2001). Effects of long-term plant Medicago sative Linn. on soil nitrogen nutrient. Chinese Journal of Ecology Agriculture, 24(4), 479–479.Google Scholar
  19. Liu, Y. Q., Jiang, L., & Sun, L. H. (2005). Changes in some defensive enzyme activity induced by the piercing-sucking of brown plant hopper in rice. Journal of Plant Physiology and Molecular Biology, 31(6), 643–650.Google Scholar
  20. Luo, X. Y., Feng, C. J., & Li, H. (2004). Study on changes of SOD and proline activity during low temperature stress on Medicago sativa L. cv. Zhaodong. Grassland of China, 26(04), 79–80.Google Scholar
  21. Mao, G. L., & Xu, X. (2005). Studies on in vivo selection of salt-tolerant mutant of Lycium barbarum L. and its physiological and biochemical characteristics. Acta Bot. Boreal. -Occident.Sin, 25(2), 275–280.Google Scholar
  22. McKay, H. M., & Mason, W. L. (1991). Physiological indicators of tolerance to cold storage in Sitka spruce and Douglas-fir seedlings. Canadian Journal of Forest Research, 21(6), 890–901.Google Scholar
  23. Mohammad, H. S. M., Nematollah, E., Mohammad, M. A., Mostafa, A., Mostafa, A., & Mohammad, P. (2017). Molecular and physiological responses of Iranian Perennial ryegrass as affected by Trinexapac ethyl, Paclobutrazol and Abscisic acid under drought stress. Plant Physiology and Biochemistry, 111, 129–143.CrossRefGoogle Scholar
  24. Rasouli, S., Farkhondeh, M. M., Jafari, H. R., Suffling, R., Kiabi, B., & Yavari, A. R. (2012). Assessment of Ecological integrity in a landscape context using the Miankale peninsula of Northern Iran. International Journal of Environmental Research, 6(2), 443–450.Google Scholar
  25. Shao, Y. R., Xu, J. X., & Xue, L. (2013). Effects of low temperature stress on physiological-biochemical indexes and photosynthetic characteristics of seedlings of four plant species. Acta Ecologica Sinica, 33(14), 4237–4247.CrossRefGoogle Scholar
  26. Shi, X. H., & Chen, Z. Y. (1996). Low temperature stress on SOD and its isoenzyme in detached citrus leaves. Bulletin of Botanical Researvh, 4, 384–386.Google Scholar
  27. Shi, Y. H., Zhang, W. Y., & Yu, X. D. (2009). Effect of photoperiod on SOD and POD activities in alfalfa. Grassland and Turf, 01, 74–77.Google Scholar
  28. Shi, X. H., Chen, Z. Y., Yang, H. Q., & Chen, A. (1996). Low temperature stress on SOD and its isoenzyme in detached citrus leaves. Acta Horticulturae Sinica, 23(4), 386–398.Google Scholar
  29. Wang, X. K. (2010). Experiment principles and techniques of plantphysiology and biochemistry. Beijing: Higher Education Press. Google Scholar
  30. Wang, W., & Li, Z. J. (2009). The study on changing regularity of several cold resistance indexes of alfalfa under low temperature stress, 6, 23-26. Northeast Normal University. (doctoral thesis) Google Scholar
  31. Wang, R. Y., Ren, Y. S., & Yue, W. B. (2006). Effect of low temperature stress on the survival and physiological and biochemical indexes of alfalfa seedlings. ACIA Laser Biology Snice, 15(4), 342–348.Google Scholar
  32. Wei, W. L., Cheng, J. M., Gao, Y., & Liu, W. (2010). Effects of different site conditions on alfalfa field and path analysis in arid area of Northern Weihe River Basin. Bulletin of Soil and Water Conservation, 30(5), 73–78.Google Scholar
  33. Wu, J. H., Yang, L., & Sun, G. R. (2004). Generation of activated oxygen and change of cell defense enzyme activity in leaves of maize seedling under the stress of low temperature. Bulletin of Botanical Research, 24(04), 456–459.Google Scholar
  34. Wu, H., Zhang, J. S., & Shi, J. Y. (2013). Physiological responses of cotton seedlings under low temperature stress. Acta Bot. Boreal. -Occident.Sin, 33(01), 74–82.Google Scholar
  35. Xu, Z. H., Guo, D. P., Yu, L. Q., Zhao, M., Zhang, X., Li, D., Zheng, K. L., & Ye, Y. L. (2003). Molecular biological study on the action mechanism of rice allelochemicals against weeds. Chinese Journal of Applied Ecology, 14(5), 829–833.Google Scholar
  36. Yang, X. J. (2006). Evaluation of cold resistance of alfalfa and its physiological responses to low temperature stress in autumn and winter. Beijing Forestry University. (doctoral thesis) Google Scholar
  37. Yang, Y. H., Jiang, P. A., & Luo, M. (2005). Effects of planting Medicago sativa L on soil fertility. Arid Land Geography, 28(2), 248–251.Google Scholar
  38. Ye, Y. X., Jin, J., & Qin, F. J. (2009). Effects of low temperature stress on superoxide dismutase activity of wheat, corn, radish seedlings. Chinese Agricultural Science Bulletin, 25(23), 244–248.Google Scholar
  39. Yu, J., Cang, J., Li, Y. P., Huang, R., Lu, Q. W., Wang, X. T., Liu, L. J., Xu, Q. H., & Zhang, K. J. (2016). Salicylic acid-induced antioxidant protection against low temperature in cold-hardy winter wheat. Acta Physiologiae Plantarum, 38(11), 1–10.Google Scholar
  40. Yuan, Y. S., Hu, Y., & Li, Y. X. (2014). Effects of simulated acid rain stress on growth and physiological characteristics in leaves of Acanthopanan trifoliatus (L.) Merr. Journal of Sichuan Agricultural University, 32(01), 28–33.Google Scholar
  41. Zhang, W. H., & Hu, T. M. (2002). Effect of low temperature stress on physiological of warm season turfgrass Dichondra repens forst. Journal of Jilin Agricultural University, 24(5), 39–41.Google Scholar
  42. Zhou, X. Q. (2004). Acid rain effect on the physiological system of Casuarina equisetifolia seedlings. Journal of Mountain Agriculture and Biology, 23(3), 210–214.Google Scholar
  43. Zou, Q. (2000). Experimental instruction of plant physiology. Bei Jing: China Agriculture Press.Google Scholar

Copyright information

© Springer International Publishing AG 2017

Authors and Affiliations

  • Guozhang Bao
    • 1
    Email author
  • Qi Ao
    • 2
  • Qiqi Li
    • 1
  • Yishu Bao
    • 1
  • Yue Zheng
    • 1
  • Xiaoxia Feng
    • 1
  • Xuemei Ding
    • 3
  1. 1.Key Laboratory of Groundwater Resources and Environment of the Ministry of Education, College of Environment and ResourcesJilin UniversityChangchunChina
  2. 2.College of Environmental Science and EngineeringPeking UniversityBeijingChina
  3. 3.College of Animal ScienceJilin UniversityChangchunChina

Personalised recommendations