Real-time PCR quantification of arbuscular mycorrhizal fungi: does the use of nuclear or mitochondrial markers make a difference?
Jazyk angličtina Země Německo Médium print-electronic
Typ dokumentu časopisecké články
PubMed
28569349
DOI
10.1007/s00572-017-0777-9
PII: 10.1007/s00572-017-0777-9
Knihovny.cz E-zdroje
- Klíčová slova
- Arbuscular mycorrhizal fungi, Isolate discrimination, Microsymbiont screening, Mitochondrial DNA, Molecular genetic quantification, Nuclear ribosomal DNA, PLFA, Real-time PCR,
- MeSH
- buněčné jádro genetika MeSH
- DNA fungální genetika MeSH
- Glomeromycota genetika MeSH
- kořeny rostlin mikrobiologie MeSH
- kvantitativní polymerázová řetězová reakce * MeSH
- Medicago truncatula mikrobiologie MeSH
- mitochondriální DNA genetika MeSH
- mykorhiza genetika MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- DNA fungální MeSH
- mitochondriální DNA MeSH
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.
Institute of Botany The Czech Academy of Sciences Zámek 1 Průhonice 252 43 Czech Republic
Institute of Microbiology The Czech Academy of Sciences Vídeňská 1083 Prague 142 20 Czech Republic
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