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Analysis of Arbuscular Mycorrhizal Fungal Communities by Terminal Restriction Fragment Length Polymorphism (TRFLP)

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Arbuscular Mycorrhizal Fungi

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2146))

Abstract

Terminal restriction fragment length polymorphism (TRFLP) approaches enable the detection and identification of microbial taxa into samples coming from root or soil material DNA extraction. The low taxonomic diversity of arbuscular mycorrhizal (AM) fungi makes this technique a cheap and adequate method for fingerprinting their communities. Here, I describe the TRFLP database approach, a version of the technique in which the AM fungal taxa present in the sample pool is identified for, later, match their presence in the different samples contained in the experiment. A final AM fungal operational taxonomic unit x sample presence–absence matrix is obtained, which allows the subsequent multivariate statistical analysis of the experimental results.

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References

  1. van der Heijden MGA, Martin FM, Selosse MA, Sanders IR (2015) Mycorrhizal ecology and evolution: the past, the present, and the future. New Phytol 205:1406–1423

    Article  Google Scholar 

  2. Courtney KC, Bainard LD, Sikes BA et al (2012) Determining a minimum detection threshold in terminal restriction fragment length polymorphism analysis. J Microbiol Methods 88:14–18

    Article  CAS  Google Scholar 

  3. Martínez-García LB, Richardson SJ, Tylianakis JM et al (2015) Host identity is a dominant driver of mycorrhizal fungal community composition during ecosystem development. New Phytol 205:1565–1576

    Article  Google Scholar 

  4. Varela-Cervero S, López-García Á, Barea JM, Azcón-Aguilar C (2016) Spring to autumn changes in the arbuscular mycorrhizal fungal community composition in the different propagule types associated to a Mediterranean shrubland. Plant Soil 26:107–120

    Article  Google Scholar 

  5. López-García Á, Varela-Cervero S, Vasar M et al (2017) Plant traits determine the phylogenetic structure of arbuscular mycorrhizal fungal communities. Mol Ecol 26:6948–6959

    Article  Google Scholar 

  6. Dickie IA, FitzJohn RG (2007) Using terminal restriction fragment length polymorphism (T-RFLP) to identify mycorrhizal fungi: a methods review. Mycorrhiza 17:259–270

    Article  CAS  Google Scholar 

  7. Koch AM, Kuhn G, Fontanillas P et al (2004) High genetic variability and low local diversity in a population of arbuscular mycorrhizal fungi. Proc Natl Acad Sci U S A 101:2369–2374

    Article  CAS  Google Scholar 

  8. López-García Á, Palenzuela J, Barea JM, Azcón-Aguilar C (2014) Life-history strategies of arbuscular mycorrhizal fungi determine succession into roots of Rosmarinus officinalis L., a characteristic woody perennial plant species from Mediterranean ecosystems. Plant Soil 379:247–260

    Article  Google Scholar 

  9. Lee J, Lee S, Young JPW (2008) Improved PCR primers for the detection and identification of arbuscular mycorrhizal fungi. FEMS Microbiol Ecol 65:339–349

    Article  CAS  Google Scholar 

  10. Schloss PD, Westcott SL, Ryabin T et al (2009) Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl Environ Microbiol 75:7537–7541

    Article  CAS  Google Scholar 

  11. Kumar S, Stecher G, Li M et al (2018) MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol 35:1547–1549

    Article  CAS  Google Scholar 

  12. Huson DH, Bryant D (2006) Application of phylogenetic networks in evolutionary studies. Mol Biol Evol 23:254–267

    Article  CAS  Google Scholar 

  13. Collins RE, Rocap G (2007) REPK: an analytical web server to select restriction endonucleases for terminal restriction fragment length polymorphism analysis. Nucleic Acids Res 35:W58–W62

    Article  Google Scholar 

  14. Fitzjohn RG, Dickie IA (2007) TRAMPR: AN R package for analysis and matching of terminal-restriction fragment length polymorphism (TRFLP) profiles. Mol Ecol Notes 7:583–587

    Article  CAS  Google Scholar 

  15. Oksanen J, Blanchet FG, Kindt R et al (2011) Vegan: community ecology package. R package version 2.0-1. R Project. http://CRAN.R-project.org/package=vegan

  16. Lekberg Y, Gibbons SM, Rosendahl S (2014) Will different OTU delineation methods change interpretation of arbuscular mycorrhizal fungal community patterns? New Phytol 202:1101–1104

    Article  Google Scholar 

  17. Huson DH, Rupp R, Scornavacca C (2011) Phylogenetic networks: concepts, algorithms and applications. Cambridge University Press, Cambridge

    Google Scholar 

  18. Öpik M, Vanatoa A, Vanatoa E et al (2010) The online database MaarjAM reveals global and ecosystemic distribution patterns in arbuscular mycorrhizal fungi (Glomeromycota). New Phytol 188:223–241

    Article  Google Scholar 

  19. Kohout P, Sudova R, Janoušková M et al (2014) Comparison of commonly used primer sets for evaluating arbuscular mycorrhizal fungal communities: is there a universal solution? Soil Biol Biochem 68:482–493

    Article  CAS  Google Scholar 

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

    Article  Google Scholar 

  21. Ohsowski BM, Zaitsoff PD, Öpik M, Hart MM (2014) Where the wild things are: looking for uncultured Glomeromycota. New Phytol 204:171–179

    Article  Google Scholar 

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Acknowledgments

The author was supported by Spanish government under the Plan Nacional de I+D+I (project CGL2015-69118-C2-2-P-COEXMED-II) and University of Jaén (Spain) under the Plan 6-UJA postdoctoral fellowship.

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Correspondence to Álvaro López-García .

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López-García, Á. (2020). Analysis of Arbuscular Mycorrhizal Fungal Communities by Terminal Restriction Fragment Length Polymorphism (TRFLP). In: Ferrol, N., Lanfranco, L. (eds) Arbuscular Mycorrhizal Fungi. Methods in Molecular Biology, vol 2146. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0603-2_10

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  • DOI: https://doi.org/10.1007/978-1-0716-0603-2_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0602-5

  • Online ISBN: 978-1-0716-0603-2

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