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Real-time PCR quantification of arbuscular mycorrhizal fungi: does the use of nuclear or mitochondrial markers make a difference?
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SYSNO ASEP 0482296 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Real-time PCR quantification of arbuscular mycorrhizal fungi: does the use of nuclear or mitochondrial markers make a difference? Author(s) Voříšková, A. (CZ)
Jansa, Jan (MBU-M) RID, ORCID
Püschel, David (MBU-M) RID, ORCID
Krüger, M. (DE)
Cajthaml, Tomáš (MBU-M) RID, ORCID
Vosátka, M. (CZ)
Janoušková, M. (CZ)Source Title Mycorrhiza. - : Springer - ISSN 0940-6360
Roč. 27, č. 6 (2017), s. 577-585Number of pages 9 s. Language eng - English Country DE - Germany Keywords Arbuscular mycorrhizal fung ; Real-time PCR ; PLFA Subject RIV EE - Microbiology, Virology OECD category Microbiology R&D Projects GA15-05466S GA ČR - Czech Science Foundation (CSF) Institutional support MBU-M - RVO:61388971 UT WOS 000405925100005 EID SCOPUS 85020123571 DOI 10.1007/s00572-017-0777-9 Annotation Root colonization by arbuscular mycorrhizal fungi (AMF) can be quantified by different approaches. We compared two approaches that enable discrimination of specific AMF taxa and are therefore emerging as alternative to most commonly performed microscopic quantification of AMF in roots: quantitative real-time PCR (qPCR) using markers in nuclear ribosomal DNA (nrDNA) and mitochondrial ribosomal DNA (mtDNA). In a greenhouse experiment, Medicago truncatula was inoculated with four isolates belonging to different AMF species (Rhizophagus irregularis, Claroideoglomus claroideum, Gigaspora margarita and Funneliformis mosseae). The AMF were quantified in the root samples by qPCR targeted to both markers, microscopy and contents of AMF-specific phospholipid fatty acids (PLFA). Copy numbers of nrDNA and mtDNA were closely related within all isolates, however, the slopes and intercepts of the linear relationships significantly differed among the isolates. Across all isolates, a large proportion of variance in nrDNA copy numbers was explained by root colonization intensity or contents of AMF-specific PLFA, while variance in mtDNA copy numbers was mainly explained by differences among AMF isolates. We propose that the encountered inter-isolate differences in the ratios of mtDNA and nrDNA copy numbers reflect different physiological states of the isolates. Our results suggest that nrDNA is a more suitable marker region than mtDNA for the quantification of multiple AMF taxa as its copy numbers are better related to fungal biomass across taxa than are copy numbers of mtDNA. Workplace Institute of Microbiology Contact Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Year of Publishing 2018
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