Effects of Arbuscular Mycorrhizal Fungi on Metals Uptake, Physiological and Biochemical Response of Medicago Sativa L. with Increasing Zn and Cd Concentrations in Soil


The effect of mycorrhizal symbiosis on metal accumulation and plant tolerance are not commonly studied in medicinal plants under metal stress. The objective of this study was to assess the impact of mycorrhiza on alfalfa plants with the increase of Zn and Cd toxicity. The experiment was conducted under controlled laboratory conditions. Zinc (Zn) and cadmium (Cd) uptake, some biochemical and physiological parameters were studied in eight-week-old alfalfa plants in response to inoculation or not with arbuscular mycorrhizal fungi (AMF) and with the increase of Zn (0, 100, 300, 900 mg·kg-1) and Cd concentrations (0, 100, 300, 600 mg·kg-1) in soil. The results showed that mycorrhizal (M) plants exhibited tolerance to Zn and Cd up to 300 mg·kg-1 in comparison to non-mycorrhizal (NM) plants which exhibited a significant growth reduction at the same soil Zn and Cd level. M inoculation reduced the Zn and Cd accumulation in shoot and showed higher Zn and Cd contents in roots which showed a different Zn and Cd distribution in AMF associated or non-associated plants. Mycorrhizal plants increased phosphorus (P) contents at all Zn and Cd concentrations except the highest (600 and 900 mg·kg-1) leading significant alterations in biochemical contents such as proline, antioxidant enzymes in leaves and also in nutrients (N, P, K, Cu, Ni, Fe, Mn). Zn and cadmium toxicity cause to increase the proline content in shoot of NM plants, however, proline contents are lower in M plants. Results confirmed that AMF protected alfalfa plants against Zn and Cd toxicity. Mycorrhizal colonization was able to form an efficient symbiosis with alfalfa plants in moderately contaminated Zn and Cd soils (300 mg·kg-1) and play an important role in food quality and safety.

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Kanwal, S. , Bano, A. and Malik, R. (2015) Effects of Arbuscular Mycorrhizal Fungi on Metals Uptake, Physiological and Biochemical Response of Medicago Sativa L. with Increasing Zn and Cd Concentrations in Soil. American Journal of Plant Sciences, 6, 2906-2923. doi: 10.4236/ajps.2015.618287.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Wren, C.D., Harris, S. and Harttrup, N. (1995) Ecotoxicology of Mercury and Cadmium. In: Hoffman, D.J., Rattner, G.A., Burton Jr., A. and Cairns Jr., J., Eds., Handbook of Ecotoxicology, Lewis Publishers, Boca Raton, 392-423.
[2] Nan, Z.R., Zhao, C.Y., Li, J.J., Chen, F.H. and Sun, W. (2002) Relations between Soil Properties and Selected Heavy Metal Concentrations in Spring Wheat (Triticum aestivum L.) Grown in Contaminated Soils. Water, Air, and Soil Pollution, 133, 205-213.
[3] Chaney, R.L., Ryan, J.A., Li, Y.M., Welch, R.M. and Reeves, P.G. (1996) Phytoavailability and Bio-Availability in Risk Assessment for Cd in Agricultural Environments. In: Sources of Cadmium in the Environments, Organization for Economic Co-Operation and Development (OECD) Publications Service, Paris, 49-78.
[4] Kabata-Pendias, A. and Pendias, H. (2001) Trace Elements in Soils and Plants. 3rd Edition, CRC Press, Boca Raton.
[5] Khan, A.G. (2005) Role of Soil Microbes in the Rhizospheres of Plants Growing on Trace Metal Contaminated Soils in Phytoremediation. Journal of Trace Elements in Medicine and Biology, 18, 355-364.
[6] Heggo, A., Angle, J.S. and Chaney, R.L. (1990) Effects of Vesicular-Arbuscular Mycorrhizal Fungi on Heavy Metal Uptake by Soybeans. Soil Biology and Biochemistry, 22, 865-869.
[7] Harleym, J.L. and Smith, S.E. (1983) Mycorrhizal Symbiosis. Academic Press, Toronto.
[8] Rivera-Becerril, F., Calantzis, C., Turnau, K., Caussanel, J.P., Belimov, A.A., Gianinazzi, S. and Gianinazzi-Pearson, V. (2002) Cadmium Accumulation and Buffering of cadmium-Induced Stress by Arbuscular Mycorrhiza in Three Pisum sativum L. Genotypes. Journal of Experimental Botany, 53, 1177-1185.
[9] Hutchinson, J.J. (2001) Assessing the Bioavailability of Cadmium in Soils and Implications for Phytoremediation. PhD Thesis, University of Nottingham, Nottingham.
[10] Hildebrandt, U., Regvar, M. and Bothe, H. (2007) Arbuscular Mycorrhiza and Heavy Metal Tolerance. Phytochemistry, 68, 139-146.
[11] Smith, S.E. and Read, D.J. (2008) Mycorrhizal Symbiosis. 3rd Edition, Academic Press, London.
[12] Hildebrandt, U., Kaldorf, M. and Bothe, H. (1999) The Zinc Violet and Its Colonization by Arbuscular Mycorrhizal Fungi. Journal of Plant Physiology, 154, 709-717.
[13] Schweiger, P. and Jakobsen, I. (2000) Laboratory and Field Methods for Measurement of Hyphal Uptake of Nutrients in Soil. Plant and Soil, 226, 237-244.
[14] Muleta, D., Assefa, F., Nemomissa, S. and Granhall, U. (2007) Composition of Coffee Shade Tree Species and Density of Indigenous Arbuscular Mycorrhizal Fungi (AMF) Spores in Bonga Natural Coffee Forest, Southwestern Ethiopia. Forest Ecology and Management, 241, 145-154.
[15] Koske, R.E. and Gemma, J.N. (1992) Fungal Reactions to Plants Prior to Mycorrhizal Formation. In: Allen, M.F., Ed., Mycorrhizal Functioning: An Integrative Plant Fungal Process, Chapman & Hall, New York, 3-27.
[16] Giovannetti, M. and Mosse, B. (1980) An Evaluation of Techniques for Measuring Vesicular-Arbuscular Mycorrhizal Infection Roots. New Phytologist, 84, 489-500.
[17] Ryan, J., Estefan, G. and Rashid, A. (2001) Soil and Plant Analysis Laboratory Manual. 2nd edition, Syrian Arab Republic ICARDA, Aleppo.
[18] Van Schouwenberg, J.C.H. and Walinge, I. (1973) Methods of Analysis for Plant Material. Agriculture University, Wageningen.
[19] Hiscox, J.D. and Israelstam, G.F. (1979) A Method for the Extraction of Chlorophyll from Leaf Tissue without Maceration. Canadian Journal of Botany, 57, 1332-1334.
[20] Dubois, M., Gilles, K.A., Hamilton, J.K., Rebers, P.A. and Ith, F.S. (1956) Calorimetric Method for Determination of Sugars and Related Substances. Analytical Chemistry, 28, 350-356.
[21] Bates, L.S., Waldren, R.P. and Teare, I.D. (1973) Rapid Determination of Free Proline for Water Stress Studies. Plant and Soil, 39, 205-207.
[22] Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.J. (1951) Protein Measurement with the Folin Phenol Reagent. Journal of Biological Chemistry, 193, 265-275.
[23] Beauchamp, C.O. and Fridovich, I. (1971) Superoxide Dismutase: Improved Assays and an Assay Applicable to Acrylamide Gels. Analytical Biochemistry, 44, 276-287.
[24] Gorin, N. and Heidema, F.T. (1976) Peroxidase Activity in Golden Delicious Apples as a Possible Parameter of Ripening and Senescence. Journal of Agricultural and Food Chemistry, 24, 200-201.
[25] Goel, A., Goel, A.K. and Sheoran, I.S. (2003) Changes in Oxidative Stress Enzymes during Artificial Aging in Cotton (Gossypium hirsutum L.) Seeds. Journal of Plant Physiology, 160, 1093-1100.
[26] Nakano, Y. and Asada, K. (1981) Hydrogen Peroxide Is Scavenged by Ascorbate Peroxidase in Spinach Chloroplasts. Plant and Cell Physiology, 22, 867-880.
[27] Or1owska, E., Przyby1owicz, W., Orlowski, D., Turnau, K. and Mesjasz-Przyby1owicz, J. (2011) The Effect of Mycorrhiza on the Growth and Elemental Composition of Ni-Hyperaccumulating Plant Berkheya coddii Roessler. Environmental Pollution, 159, 3730-3738.
[28] Malekzadeh, E., Alikhani, A.H., Savaghebi Fioozabadi, R.G. and Zarei, M. (2011) Influence of Arbuscular Mycorrhizal Fungi and an Improving Growth Bacterium on Cd uptake and maize growth in Cd-polluted Soils. Spanish Journal of Agricultural Research, 9, 1213-1223.
[29] Gupta, M.L., Prasad, A., Ram, M. and Kumal, S. (2002) Effect of the AM Fungus G. fasciculatum on the Essential Oil Yield Condition Related Characters and Nutrient Acquisition in the Crops of Different Cultivars of Menthol Mint (Mentha arvensis) under Field Conditions. Bioresource Technology, 81, 77-79.
[30] Vivas, A., Voros, A., Biro, B., Barea, J.M., Ruiz-Lozano, J.M. and Azcon, R. (2003) Beneficial Effects of Indigenous Cd-Tolerant and Cdsensitive Glomus mosseae Associated with a Cd-Adapted Strain of Brevibacillus sp in Improving Plant Tolerance to Cd Contamination. Applied Soil Ecology, 24, 177-186.
[31] Schenck, N.C. and Hinson, K. (1973) Response of Nodulating and Non-Nodulating Soybeans to a Species of Endogone Mycorrhiza. Agronomy Journal, 65, 849-850.
[32] Chen, B.D., Zhu, Y.G., Duan, J., Xiao, X.Y. and Smith, S.E. (2007) Effects of the Arbuscular Mycorrhizal Fungus Glomus mosseae on Growth and Metal Uptake by Four Plant Species in Copper Mine Tailings. Environmental Pollution, 147, 374-380.
[33] Wang, X.K., Manning, W.J., Feng, Z.W. and Zhu, Y.G. (2007) Ground Level Ozone in China: Distribution and Effects on Crop Yields. Environmental Pollution, 147, 394-400.
[34] El-Kherbawy, M., Angle, J.S. and Chaney, R.L. (1989) Soil pH, Rhizobia and Vesicular-Arbuscular Mycorrhizae Inoculation Effects on Growth and Heavy Metal Uptake of Alfalfa (Medicago sativa L.). Biology and Fertility of Soils, 8, 61-65.
[35] Rufyikiri, G., Huysmans, L., Wannijn, J., Van Hees, M., Leyval, C. and Jakobsen, I. (2004) Arbuscular Mycorrhizal Fungi Can Decrease the Uptake of Uranium by Subterranean Clover Grown at High Levels of Uranium in Soil. Environmental Pollution, 130, 427-436.
[36] Smith, S.E. and Smith, A.F. (2011) Roles of Arbuscular Mycorrhizas in Plant Nutrition and Growth: New Paradigms from Cellular to Ecosystem Scales. Annual Review of Plant Biology, 62, 227-250.

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