Quantitative analysis of phytopathogenic ascomycota on leaves of pedunculate oaks (Quercus roburL.) by real-time PCR

Autor:	Wolfgang Zimmer, Thomas Heuser
Rok vydání:	2002
Předmět:	Ramularia biology Ascomycota Hypha Microorganism fungi Fungus biology.organism_classification DNA Ribosomal Polymerase Chain Reaction Microbiology Plant Leaves Quercus robur Quercus Botany Genetics Microsphaera Cloning Molecular DNA Fungal Molecular Biology Plant Diseases Cladosporium
Zdroj:	FEMS Microbiology Letters. 209:295-299
ISSN:	1574-6968 0378-1097
DOI:	10.1111/j.1574-6968.2002.tb11147.x
Popis:	Leaves of oak trees are often infected by various pathogenic fungi. As it is difficult to remove such organisms quantitatively from the leaf surface and as it is often impossible to grow these organisms independently from their host, there are almost no molecular data available from these oak leaf specific pathogens. For the quantitative removal of the microorganisms a procedure was developed combining a wax and microorganism freezing method with a DNA extraction technique. For the development of a species specific detection, DNA of pathogenic filamentous fungi was isolated from hyphae of the upper leaf surface of Quercus robur. Three different species could be identified as (i) Cladosporium sp., (ii) Ramularia sp. and (iii) Microsphaera alphitoides by amplifying and sequencing an 18S-28S segment of their rDNA. For the final quantification a real-time PCR protocol was established allowing the species specific quantification of the three pathogenic filamentous fungi. The whole procedure was successfully applied to quantify the amount of the three species on oak leaves collected in autumn.
Databáze:	OpenAIRE
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