Your browser doesn't support javascript.
loading
Development of a Real-Time PCR assay for quantitative assessment of uncultured freshwater zoosporic fungi.
Lefèvre, Emilie; Jobard, Marlène; Venisse, Jean-Stéphane; Bec, Alexandre; Kagami, Maiko; Amblard, Christian; Sime-Ngando, Télesphore.
Afiliação
  • Lefèvre E; LMGE, Laboratoire 'Microorganismes: Génome & Environnement', UMR CNRS 6023, Université Blaise Pascal (Clermont-Ferrand II), BP 80026, 63171 Aubière Cedex, France.
J Microbiol Methods ; 81(1): 69-76, 2010 Apr.
Article em En | MEDLINE | ID: mdl-20153382
Recently, molecular environmental surveys of the eukaryotic microbial community in lakes have revealed a high diversity of sequences belonging to uncultured zoosporic fungi. Although they are known as saprobes and algal parasites in freshwater systems, zoosporic fungi have been neglected in microbial food web studies. Recently, it has been suggested that zoosporic fungi, via the consumption of their zoospores by zooplankters, could transfer energy from large inedible algae and particulate organic material to higher trophic levels. However, because of their small size and their lack of distinctive morphological features, traditional microscopy does not allow the detection of fungal zoospores in the field. Hence, quantitative data on fungal zoospores in natural environments is missing. We have developed a quantitative PCR (qPCR) assay for the quantification of fungal zoospores in lakes. Specific primers were designed and qPCR conditions were optimized using a range of target and non-target plasmids obtained from previous freshwater environmental 18S rDNA surveys. When optimal DNA extraction protocol and qPCR conditions were applied, the qPCR assay developed in this study demonstrated high specificity and sensitivity, with as low as 100 18S rDNA copies per reaction detected. Although the present work focuses on the design and optimization of a new qPCR assay, its application to natural samples indicated that qPCR offers a promising tool for quantitative assessment of fungal zoospores in natural environments. We conclude that this will contribute to a better understanding of the ecological significance of zoosporic fungi in microbial food webs of pelagic ecosystems.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Reação em Cadeia da Polimerase / Água Doce / Fungos Tipo de estudo: Diagnostic_studies / Evaluation_studies Idioma: En Ano de publicação: 2010 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Reação em Cadeia da Polimerase / Água Doce / Fungos Tipo de estudo: Diagnostic_studies / Evaluation_studies Idioma: En Ano de publicação: 2010 Tipo de documento: Article