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

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 s...

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Bibliographic Details
Published in:FEMS microbiology letters 2002-04, Vol.209 (2), p.295-299
Main Authors: Heuser, T, Zimmer, W
Format: Article
Language:English
Subjects:
Ascomycota - genetics
Ascomycota - isolation & purification
autumn
Biological and medical sciences
Cladosporium
Cloning, Molecular
Deoxyribonucleic acid
DNA
DNA, Fungal - analysis
DNA, Ribosomal - genetics
Erysiphe alphitoides
Freezing
Fundamental and applied biological sciences. Psychology
Fungal plant pathogens
Fungi
Generalities. Techniques
Hyphae
Leaves
Microbiology
Microorganisms
Microsphaera alphitoides
Mycology
Oak
Pathogenicity, host-agent relations, miscellaneous strains, epidemiology
pathogens
Phytopathology. Animal pests. Plant and forest protection
Plant Diseases - microbiology
Plant Leaves - microbiology
Polymerase Chain Reaction
Powdery mildew
Quantitative analysis
quantitative polymerase chain reaction
Quercus - microbiology
Quercus robur
Ramularia
Real time
Real‐time polymerase chain reaction
ribosomal DNA
Species
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Summary: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.
ISSN:0378-1097
1574-6968
DOI:10.1111/j.1574-6968.2002.tb11147.x