title: Evaluation of culture-based techniques and 454 pyrosequencing for the analysis of fungal diversity in potting media and organic fertilizers.
abstract: OBJECTIVE
  Potting media and organic fertilizers (OFs) are commonly used in agricultural systems. However, there is a lack of studies on the efficiency of culture-based techniques in assessing the level of fungal diversity in these products. A study was conducted to investigate the efficiency of seven culture-based techniques and pyrosequencing for characterizing fungal diversity in potting media and OFs.
  
  
  RESULTS
  Fungal diversity was evaluated using serial dilution, direct plating and baiting with carrot slices, potato slices, radish seeds, cucumber seeds and cucumber cotyledons. Identity of all the isolates was confirmed on the basis of the internal transcribed spacer region of the ribosomal RNA (ITS rRNA) sequence data. The direct plating technique was found to be superior over other culture-based techniques in the number of fungal species detected. It was also found to be simple and the least time consuming technique. Comparing the efficiency of direct plating with 454 pyrosequencing revealed that pyrosequencing detected 12 and 15 times more fungal species from potting media and OFs respectively. Analysis revealed that there were differences between potting media and OFs in the dominant phyla, classes, orders, families, genera and species detected. Zygomycota (52%) and Chytridiomycota (60%) were the predominant phyla in potting media and OFs respectively.
  
  
  CONCLUSIONS
  The superiority of pyrosequencing over cultural methods could be related to the ability to detect obligate fungi, slow growing fungi and fungi that exist at low population densities.
  
  
  CONCLUSIONS
  The evaluated methods in this study, especially direct plating and pyrosequencing, may be used as tools to help detect and reduce movement of unwanted fungi between countries and regions.
authors.affiliation: Department of Crop Sciences, College of Agricultural and Marine Sciences, Sultan Qaboos University, Al Khoud, Oman.
authors.forename: A M
authors.initials: AM
authors.lastname: Al-Sadi
authors.affiliation: The Research Council, Al Athibah, Oman.
authors.forename: S S
authors.initials: SS
authors.lastname: Al-Mazroui
authors.affiliation: Department of Life Sciences, New University of Lisbon, Caparica, Portugal.
authors.forename: A J L
authors.initials: AJ
authors.lastname: Phillips
mesh_terms: D044822:Biodiversity; D046508:Culture Techniques / Q000379:methods*; D005308:Fertilizers / Q000032:analysis / Q000382:microbiology*; D005658:Fungi / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D012988:Soil Microbiology
chemical_list: D005308:Fertilizers
keywords: Ascomycota; Basidiomycota; Chytridiomycetes; Sordariomycetes; baiting; next generation sequencing; plating