Microbial Ecology

, 55:45

Influence of Salinity on the In Vitro Development of Glomus intraradices and on the In Vivo Physiological and Molecular Responses of Mycorrhizal Lettuce Plants

  • Farzad Jahromi
  • Ricardo Aroca
  • Rosa Porcel
  • Juan Manuel Ruiz-Lozano


Increased salinization of arable land is expected to have devastating global effects in the coming years. Arbuscular mycorrhizal fungi (AMF) have been shown to improve plant tolerance to abiotic environmental factors such as salinity, but they can be themselves negatively affected by salinity. In this study, the first in vitro experiment analyzed the effects of 0, 50, or 100 mM NaCl on the development and sporulation of Glomus intraradices. In the second experiment, the effects of mycorrhization on the expression of key plant genes expected to be affected by salinity was evaluated. Results showed that the assayed isolate G. intraradices DAOM 197198 can be regarded as a moderately salt-tolerant AMF because it did not significantly decrease hyphal development or formation of branching absorbing structures at 50 mM NaCl. Results also showed that plants colonized by G. intraradices grew more than nonmycorrhizal plants. This effect was concomitant with a higher relative water content in AM plants, lower proline content, and expression of Lsp5cs gene (mainly at 50 mM NaCl), lower expression of the stress marker gene Lslea gene, and lower content of abscisic acid in roots of mycorrhizal plants as compared to nonmycorrhizal plants, which suggest that the AM fungus decreased salt stress injury. In addition, under salinity, AM symbiosis enhanced the expression of LsPIP1. Such enhanced gene expression could contribute to regulating root water permeability to better tolerate the osmotic stress generated by salinity.


  1. 1.
    Adiku, G, Renger, M, Wessolek, G, Facklam, M, Hech-Bischoltz, C (2001) Simulation of dry matter production and seed yield of common beans under varying soil water and salinity conditions. Agric Water Manag 47: 55–68CrossRefGoogle Scholar
  2. 2.
    Al-Karaki, GN (2006) Nursery inoculation of tomato with arbuscular mycorrhizal fungi and subsequent performance under irrigation with saline water. Sci Hortic 109: 1–7CrossRefGoogle Scholar
  3. 3.
    Al-Karaki, GN, Hammad, R, Rusan, M (2001) Response of two tomato cultivars differing in salt tolerance to inoculation with mycorrhizal fungi under salt stress. Mycorrhiza 11: 43–47CrossRefGoogle Scholar
  4. 4.
    Aroca, R, Tognoni, F, Irigoyen, JJ, Sánchez-Díaz, M, Pardossi A (2001) Different root low temperature response of two maize genotypes differing in chilling sensitivity. Plant Physiol Biochem 39: 1067–1073CrossRefGoogle Scholar
  5. 5.
    Aroca, R, Vernieri, P, Irigoyen, JJ, Sánchez-Díaz, M, Tognoni, F, Pardossi, A (2003) Involvement of abscisic acid in leaf and root of maize (Zea mays L.) in avoiding chilling-induced water stress. Plant Sci 165: 671–679CrossRefGoogle Scholar
  6. 6.
    Bago, B, Cano, C (2005) Breaking myths on arbuscular mycorrhizas in vitro biology. In: Declerck, S, Strullu, DG, Fortin, JA (Eds.) In Vitro Culture of Mycorrhizas, Springer, Berlin Heidelberg New York, pp 111–138CrossRefGoogle Scholar
  7. 7.
    Bago, B, Azcon-Aguilar, C, Goulet, A, Piche, Y (1998) Branched absorbing structure (BAS), a feature of the extrarradical mycelium of symbiotic arbuscular mycorrhizal fungi. New Phytol 139: 375–388CrossRefGoogle Scholar
  8. 8.
    Barrieu, F, Marty-Mazars, D, Thomas, D, Chaumont, F, Charbonnier, M, Marty, F (1999) Desiccation and osmotic stress increase the abundance of mRNA of the tonoplast aquaporin BobTIP26-1 in cauliflower cells. Planta 209: 77–86PubMedCrossRefGoogle Scholar
  9. 9.
    Bates, LS, Waldren, RP, Teare, ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39: 205–207CrossRefGoogle Scholar
  10. 10.
    Blig, EG, Dyer, WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37: 911–917Google Scholar
  11. 11.
    Blintsov, AN, Gussakovskaya, MA (2004) Immunochemical approach to the problem of differential determination of natural forms of abscisic acid. Biochemistry (Mosc) 69: 1099–1108CrossRefGoogle Scholar
  12. 12.
    Bray, EA (2002) Abscisic acid regulation of gene expression during water-deficit stress in the era of the Arabidopsis genome. Plant Cell Environ 25: 153–161PubMedCrossRefGoogle Scholar
  13. 13.
    Cho, K, Toler, H, Lee, J, Ownley, B, Stutz, JC, Moore, JL, Augé, RM (2006) Mycorrhizal symbiosis and response of sorghum plants to combined drought and salinity stresses. J Plant Physiol 163: 517–528PubMedCrossRefGoogle Scholar
  14. 14.
    Close, TJ (1996) Dehydrins: emergence of a biochemical role of a family of plant dehydration proteins. Physiol Plant 97: 795–803CrossRefGoogle Scholar
  15. 15.
    Cranenbrouk, S, Voets, L, Bivort, C (2005) Methodologies for in vitro cultivation of arbuscular mycorrhizal fungi with root organs. In: Declerck, S, Strullu, DG, Fortin, JA (Eds.) In Vitro Culture of Mycorrhizas, Springer, Berlin Heidelberg New York, pp 341–375CrossRefGoogle Scholar
  16. 16.
    Declerck, S, Strullu, DG, Fortin, JA (2005) In Vitro Culture of Mycorrhizas. Springer, Berlin Heidelberg New YorkGoogle Scholar
  17. 17.
    Duan, X, Newman, DS, Reiber, JM, Green, CD, Saxton, AM, Augé, RM (1996) Mycorrhizal influence on hydraulic and hormonal factors implicated in the control of stomatal conductance during drought. J Exp Bot 47: 1541–1550CrossRefGoogle Scholar
  18. 18.
    Duncan, DB (1955) Multiple range and multiple F-tests. Biometrics 11: 1–42CrossRefGoogle Scholar
  19. 19.
    Estrada-Luna, AA, Davies, FT (2003) Arbuscular mycorrhizal fungi influence water relations, gas exchange, abscisic acid and growth of micropropagated chile ancho pepper (Capsicum annuum) plantlets during acclimatization and post-acclimatization. J Plant Physiol 160: 1073–1083PubMedCrossRefGoogle Scholar
  20. 20.
    Feng, G, Zhang, FS, Li, XL, Tian, CY, Tang, C (2002) Improved tolerance of maize plants to salt stress by arbuscular mycorrhizal is related to higher accumulation of soluble sugars in roots. Mycorrhiza 12: 185–190PubMedCrossRefGoogle Scholar
  21. 21.
    Giovannetti, M, Mosse, B (1980) An evaluation of techniques for measuring vesicular–arbuscular infection in roots. New Phytol 84: 489–500CrossRefGoogle Scholar
  22. 22.
    Giri, R, Kapoor, R, Mukerji, KG (2003) Influence of arbuscular mycorrhizal fungi and salinity on growth, biomass and mineral nutrition of Acacia auriculiformis. Biol Fertil Soils 38: 170–175CrossRefGoogle Scholar
  23. 23.
    Goicoechea, N, Szalai, G, Antolín, MC, Sánchez-Díaz, M, Paldi, E (1998) Influence of arbuscular mycorrhizae and Rhizobium on free polyamines and proline levels in water-stressed alfalfa. J Plant Physiol 153: 706–711Google Scholar
  24. 24.
    Hasegawa, PM, Bressan, RA, Zhu, KL, Bohnert, HJ (2000) Plant cellular and molecular responses to high salinity. Annu Rev Plant Physiol Plant Mol Biol 51: 463–499PubMedCrossRefGoogle Scholar
  25. 25.
    Jang, JY, Kim, DG, Kim, YO, Kim, JS, Kang, H (2004) An expression analysis of a gene family encoding plasma membrane aquaporins in response to abiotic stresses in Arabidopsis thaliana. Plant Mol Biol 54: 713–725PubMedCrossRefGoogle Scholar
  26. 26.
    Jindal, V, Atwal, A, Sekhon, BS, Rattan, S, Singh, R (1993) Effect of vesicular–arbuscular mycorrhizae on metabolism of moong plants under NaCl salinity. Plant Physiol Biochem 31: 475–481Google Scholar
  27. 27.
    Johansson, I, Karlsson, M, Johansson, U, Larsson, C, Kjellbom, P (2000) The role of aquaporins in cellular and whole plant water balance. Biochim Biophys Acta 1465: 324–342PubMedCrossRefGoogle Scholar
  28. 28.
    Juniper, S, Abbott, L (1993) Vesicular–arbuscular mycorrhizas and soil salinity. Mycorrhiza 4: 45–57CrossRefGoogle Scholar
  29. 29.
    Juniper, S, Abbott, LK (2006) Soil salinity delays germination and limits growth of hyphae from propagules of arbuscular mycorrhizal fungi. Mycorrhiza 16: 371–379PubMedCrossRefGoogle Scholar
  30. 30.
    Kawasaki, S, Borchert, C, Deyholos, M, Wang, H, Brazille, S, Kawai, K, Galbraith, D, Bohnert, HJ (2001) Gene expression profiles during the initial phase of salt stress in rice. Plant Cell 13: 889–905PubMedCrossRefGoogle Scholar
  31. 31.
    Kay, R, Chau, A, Daly, M (1987) Duplication of CaMV 35S promoter sequences creates a strong enhancer for plants genes. Science 236: 1299–1302PubMedCrossRefGoogle Scholar
  32. 32.
    Kishor, PB, Hong, Z, Miao, GH, Hu, CA, Verma, DPS (1995) Overexpression of Δ1-pyrroline-5-carboxilate synthetase increases proline production and confers osmotolerance in transgenic plants. Plant Physiol 108: 1387–1394PubMedGoogle Scholar
  33. 33.
    Ludwig-Müller, J (2000) Hormonal balance in plants during colonization by mycorrhizal fungi. In: Kapulnik Y, Douds, DD (Eds.) Arbuscular Mycorrhizas: Physiology and Function, Kluwer, The Netherlands, pp 263–285Google Scholar
  34. 34.
    Luu, DT, Maurel, C (2005) Aquaporins in a challenging environment: molecular gears for adjusting plant water status. Plant Cell Environ 28: 85–96CrossRefGoogle Scholar
  35. 35.
    Mahajan, S, Tuteja, N (2005) Cold, salinity and drought stress: an overview. Arch Biochem Biophys 444: 139–158PubMedCrossRefGoogle Scholar
  36. 36.
    Marschner, H (1995) Mineral Nutrition of Higher Plant, 2nd ed. Academic, New YorkGoogle Scholar
  37. 37.
    Morgan, JM (1984) Osmoregulation and water stress in higher plants. Annu Rev Plant Physiol 33: 299–319CrossRefGoogle Scholar
  38. 38.
    Munns, R (2005) Genes and salt tolerance: bringing them together. New Phytol 167: 645–663PubMedCrossRefGoogle Scholar
  39. 39.
    Ouziad, F, Wilde, P, Schmelzer, E, Hildebrandt, U, Bothe, H (2006) Analysis of expression of aquaporins and Na+/H+ transporters in tomato colonized by arbuscular mycorrhizal fungi and affected by salt stress. Environ Exp Bot 57: 177–186CrossRefGoogle Scholar
  40. 40.
    Phillips, JM, Hayman, DS (1970) Improved procedure of clearing roots and staining parasitic and vesicular–arbuscular mycorrhizal fungi for rapid assessment of infection. Trans Br Mycol Soc 55: 159–161CrossRefGoogle Scholar
  41. 41.
    Porcel, R, Azcón, R, Ruiz-Lozano, JM (2004) Evaluation of the role of genes encoding for Δ1-pyrroline-5-carboxylate synthetase (P5CS) during drought stress in arbuscular mycorrhizal Glycine max and Lactuca sativa plants. Physiol Mol Plant Pathol 65: 211–221CrossRefGoogle Scholar
  42. 42.
    Porcel, R, Aroca, R, Azcón, R, Ruiz-Lozano, JM (2006) PIP aquaporin gene expression in arbuscular mycorrhizal Glycine max and Lactuca sativa plants in relation to drought stress tolerance. Plant Mol Biol 60: 389–404PubMedCrossRefGoogle Scholar
  43. 43.
    Ramakrishnan, B, Johri, BN, Gupta, RK (1988) Influence of the VAM fungus Glomus caledonius on free proline accumulation in water-stressed maize. Curr Sci 57: 1082–1084Google Scholar
  44. 44.
    Ramoliya, P, Patel, H, Pandey, AN (2004) Effect of salinization of soil on growth and macro- and micro-nutrient accumulation in seedlings of Salvadora persica (Salvadoraceae). For Ecol Manag 2002: 181–193CrossRefGoogle Scholar
  45. 45.
    Rosendahl, CN, Rosendahl, S (1991) Influence of vesicular-arbuscular mycorrhizal fungi (Glomus spp.) on the response of cucumber (Cucumis sativis L.) to salt stress. Environ Exp Bot 31: 313–318CrossRefGoogle Scholar
  46. 46.
    Ruiz-Lozano, JM, Azcón, R (1997) Effect of calcium application on the tolerance of mycorrhizal lettuce plants to polyethylene glycol-induced water stress. Symbiosis 23: 9–22Google Scholar
  47. 47.
    Ruiz-Lozano, JM, Azcón, R, Gómez, M (1995) Effects of arbuscular mycorrhizal Glomus species on drought tolerance: physiological and nutritional plant responses. Appl Environ Microbiol 61: 456–460PubMedGoogle Scholar
  48. 48.
    Ruiz-Lozano, JM, Azcón, R, Gómez, M (1996) Alleviation of salt stress by arbuscular–mycorrhizal Glomus species in Lactuca sativa plants. Physiol Plant 98: 767–772CrossRefGoogle Scholar
  49. 49.
    Schwartz, SH, Qin, X, Zeevaart, JAD (2003) Elucidation of the indirect pathway of abscisic acid biosynthesis by mutants, genes, and enzymes. Plant Physiol 131: 1591–1601PubMedCrossRefGoogle Scholar
  50. 50.
    Walker-Simmons, M (1987) ABA levels and sensitivity in developing wheat embryos of sprouting resistant and susceptible cultivars. Plant Physiol 84: 61–66PubMedCrossRefGoogle Scholar
  51. 51.
    Wang, W, Vinocur, B, Altman, A (2003) Plant responses to drought, salinity and extreme temperatures: toward genetic engineering for stress tolerance. Planta 218: 1–14PubMedCrossRefGoogle Scholar
  52. 52.
    Yano-Melo, AM, Saggin, OJ, Costa-Maia, L (2003) Tolerance of mycorrhized banana (Musa sp. cv. Pacovan) plantlets to saline stress. Agric Ecosyst Environ 95: 343–348CrossRefGoogle Scholar
  53. 53.
    Yoshiba, Y, Kiyosue, T, Nakashima, K, Yamaguchi-Shinozaki, K, Shinozaki, K (1997) Regulation of levels of proline as an osmolyte in plants under water stress. Plant Cell Physiol 18: 1095–1102Google Scholar
  54. 54.
    Zhang, J, Jia, W, Yang, J, Ismail, AM (2006) Role of ABA in integrating plant responses to drought and salt stresses. Field Crops Res 97: 111–119CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Farzad Jahromi
    • 1
    • 2
  • Ricardo Aroca
    • 1
  • Rosa Porcel
    • 1
  • Juan Manuel Ruiz-Lozano
    • 1
  1. 1.Departamento de Microbiología del Suelo y Sistemas SimbióticosEstación Experimental del Zaidín (CSIC)GranadaSpain
  2. 2.E. H. Graham Centre for Agricultural Innovation, School of Agricultural and Veterinary SciencesCharles Sturt UniversityWagga WaggaAustralia

Personalised recommendations