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Effects of Co-Inoculation with Arbuscular Mycorrhizal Fungi and Rhizobia on Fungal Occupancy in Chickpea Root and Nodule Determined by Real-Time PCR

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Abstract

Legume roots in nature are usually colonized with rhizobia and different arbuscular mycorrhizal fungi (AMF) species. Light microscopy that visualizes the presence of AMF in roots is not able to differentiate the ratio of each AMF species in the root and nodule tissues in mixed fungal inoculation. The purpose of this study was to characterize the dominant species of mycorrhiza in roots and nodules of plants co-inoculated with mycorrhizal fungi and rhizobial strains. Glomus intraradices (GI), Glomus mosseae (GM), their mix (GI + GM), and six Mesorhizobium ciceri strains were used to inoculate chickpea. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to assess occupancy of these fungal species in roots and nodules. Results showed that GI molecular ratio and relative density were higher than GM in both roots and nodules. These differences in molecular ratio and density between GI and GM in nodules were three folds higher than roots. The results suggested that M. ciceri strains have different effects on nodulation and mycorrhizal colonization pattern. Plants with bacterial S3 and S1 strains produced the highest root nodulation and higher fungal density in both the roots and nodules.

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References

  1. Aliasgharzadeh N, Saleh Rastin N, Towfighi H, Alizadeh A (2001) Occurrence of arbuscular mycorrhizal fungi in saline soils of the Tabriz Plain of Iran in relation to some physical and chemical properties of soil. Mycorrhiza 11:119–122

    Article  CAS  Google Scholar 

  2. Alkan N, Gadkar V, Coburn J, Yarden O, Kapulnik Y (2004) Quantification of the arbuscular mycorrhizal fungus Glomus intraradices in host tissue using real-time polymerase chain reaction. New Phytol 161:877–885

    Article  CAS  Google Scholar 

  3. Alkan N, Gadkar V, Yarden O, Kapulnik Y (2006) Analysis of quantitative interaction between two species of arbuscular fungi, Glomus mosseae and G. intraradices, by real-time PCR. Appl Environ Microbiol 72(6):4192–4199

    Article  PubMed  CAS  Google Scholar 

  4. Baird LM, Caruso KJ (1994) Development of root nodules in Phaseolus vulgaris inoculated with Rhizobium and mycorrhizal fungi. Int J Plant Sci 155:633–639

    Article  Google Scholar 

  5. Caetano-AnolleÂs G, Gresshof PM (1991) Plant genetic control of nodulation in legumes. Annu Rev Microbiol 45:345–382

    Article  Google Scholar 

  6. Chaturvedi C, Singh R (1989) Response of chickpea (Cicer arietinum L.) to inoculation with Rhizobium and VA mycorrhiza. Proc Indian Natl Sci Acad Sect B 59:443–446

    Google Scholar 

  7. Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA mini-preparation: version II. Plant Mol Biol Rep 1(14):19–21

    Article  CAS  Google Scholar 

  8. Giovannetti M, Mosse B (1980) An evaluation of technique for measuring vesicular mycorrhizal infection in roots. New Phytol 84:489–500

    Article  Google Scholar 

  9. Gollotte A, van Tuinen D, Atkinson D (2004) Diversity of arbuscular mycorrhizal fungi colonizing roots of the grass species Agrostis capillaries and Lolium perenne in a field experiment. Mycorrhiza 14:111–117

    Article  PubMed  Google Scholar 

  10. Hajiboland R, Aliasgharzadeh N, Farsad S, Poschenrieder C (2010) Colonization with arbuscular mycorrhizal fungi improves salinity tolerance of tomato (Solanum lycopersicum L.) plants. Plant Soil 331:313–327

    Article  CAS  Google Scholar 

  11. Kawaguchi M, Minamisawa K (2010) Plant–microbe communications for symbiosis. Plant Cell Physiol 51(9):1377–1380

    Google Scholar 

  12. Long SR (2001) Genes and signals in the rhizobium—legume symbiosis. Plant Physiol 125:69–72

    Article  PubMed  CAS  Google Scholar 

  13. McGonigle TP, Miller MH, Evans DG, Fairchild GL, Swan JA (1990) A new method, which gives an objective measure of colonisation of roots by vesicular-arbuscular mycorrhizal fungi. New Phytol 115:495–501

    Article  Google Scholar 

  14. Norris JR, Read DJ, Varma AK (1992) Methods in microbiology Volume 24 Techniques for the study of mycorrhiza. Academic Press, London

    Google Scholar 

  15. Oldroyd GED, Harrison MJ, Udvardi M (2005) Peace talks and trade deal. Keys to long-term harmony in legume-microbe symbiosis. Plant Physiol 137:1205–1210

    Article  PubMed  CAS  Google Scholar 

  16. Pearson JN, Abbott LK, Jasper DA (1993) Mediation of competition between two colonizing VA mycorrhizal fungi by the host plant. New Phytol 123:93–98

    Article  Google Scholar 

  17. Pivato B, Mazurier S, Lemanceau P, Siblot S, Berta G, Mougel C, van Tuinen D (2007) Medicago species affect the community composition of arbuscular mycorrhizal fungi associated with roots. New Phytol 176:197–210

    Article  PubMed  CAS  Google Scholar 

  18. Pivato B, Offre P, Marchelli S, Barbonaglia B, Mougel C, Lemanceau P, Berta G (2009) Bacterial effects on arbuscular mycorrhizal fungi and mycorrhiza development as influenced by the bacteria, fungi, and host plant. Mycorrhiza 19:81–90

    Article  PubMed  Google Scholar 

  19. Provorov NA, Borisov AY, Tikhonovich IA (2002) Developmental genetics and evolution of symbiotic structures in nitrogen-fixing nodules and arbuscular mycorrhiza. J Theo Biol 214:215–232

    Article  CAS  Google Scholar 

  20. Scheublin TR, van der Heijden MGA (2006) Arbuscular mycorrhizal fungi colonize nonfixing root nodules of several legume species. New Phytol 172:732–738

    Article  PubMed  Google Scholar 

  21. Scheublin TR, Ridway KP, Young JPW, van der Heijden MGA (2004) Nonlegumes, legumes, and root nodules harbor different arbuscular mycorrhizal fungal communities. Appl Environ Microbiol 70:6240–6246

    Article  PubMed  CAS  Google Scholar 

  22. Schüssler AD, Schwarzott D, Walker C (2001) A new fungal phylum, the Glomeromycota: phylogeny and evolution. Mycol Res 105:1413–1421

    Article  Google Scholar 

  23. Sprent JI (2001) Nodulation in legumes. Royal Botanical Gardens, Kew, United Kingdom

    Google Scholar 

  24. van Tunien D, Jacquot E, Zhao B, Pearson V (1998) Characterization of root colonization profiles by a microcosm community of arbuscular mycorrhizal fungi using 25S rDNA targeted nested PCR. Mol Ecol 7:879–887

    Article  Google Scholar 

  25. Vierheilig H, Garcia-Garrido JM, Wyss U, Piché Y (2000) Systemic suppression of mycorrhizal colonization of barley roots already colonized by AM fungi. Soil Biol Biochem 32:589–595

    Article  CAS  Google Scholar 

  26. Weaver RW, Fredrick LR (1982) Rhizobium. In: Methods of soil analysis (ed.). Part 2. Chemical and microbiological properties. Agronomy monograph, No9 2nd edn American Society of Agronomy Madison, WI. pp 1043-1070

  27. Xie ZP, Staehelin C, Vierheilig H, Wiemken A, Jabbouri S, Broughton WJ, Vögeli-Lange R, Boller T (1995) Rhizobial nodulation factors stimulate mycorrhizal colonization of nodulating and nonnodulating soybeans. Plant Physiol 108:1519–1525

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors wish to thank Dr. M. Khayam Nekoui, Dr. R. Osfoori, Dr. I. Majidi, and the personnel of Microbial Biotechnology and Biosafety, and acknowledge the Genomics Departments of Agricultural Biotechnology Research Institute of Iran (ABRII) for their support and technical assistance. This study was supported by a grant from the Agricultural Research and Education Organization of Iran (AREEO).

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Correspondence to Alireza Tavasolee.

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Tavasolee, A., Aliasgharzad, N., Salehi, G.R. et al. Effects of Co-Inoculation with Arbuscular Mycorrhizal Fungi and Rhizobia on Fungal Occupancy in Chickpea Root and Nodule Determined by Real-Time PCR. Curr Microbiol 63, 107–114 (2011). https://doi.org/10.1007/s00284-011-9951-z

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