Multiple displacement amplification, a powerful tool for molecular genetic analysis of powdery mildew fungi

Powdery mildew fungi (Erysiphales) are probably the largest group of plant pathogens that remain uncharacterized from genetic and molecular points of view, with the only exception of the powdery mildew of cereals, Blumeria graminis. Their nature as obligate biotrophic parasites and consequent inabil...

Full description

Saved in:
Bibliographic Details
Published in:Current genetics 2007-03, Vol.51 (3), p.209-219
Main Authors: Fernández-Ortuño, Dolores, Torés, Juan A, de Vicente, Antonio, Pérez-García, Alejandro
Format: Article
Language:English
Subjects:
Airborne microorganisms
Amino Acid Sequence
Ascomycota - enzymology
Ascomycota - genetics
Ascomycota - isolation & purification
Bacillus Phages - genetics
Base Sequence
Blumeria graminis
Cereals
Cucurbita - microbiology
Culture media
Cytochrome b
Cytochromes b - genetics
Deoxyribonucleic acid
DNA
DNA, Ribosomal Spacer - genetics
Epidemiology
Erysiphales
Fidelity
Fungi
Genetic analysis
Genetic markers
Genetics
Genomes
genomics
Media (culture)
Mitochondria
Molecular Sequence Data
Nucleic Acid Amplification Techniques
Parasites
Pathogens
Polymerase Chain Reaction
Population genetics
Powdery mildew
Sequence Analysis, DNA
Spacer
Systematics
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Powdery mildew fungi (Erysiphales) are probably the largest group of plant pathogens that remain uncharacterized from genetic and molecular points of view, with the only exception of the powdery mildew of cereals, Blumeria graminis. Their nature as obligate biotrophic parasites and consequent inability to grow on culture media has significantly hampered research. A common bottleneck to the molecular genetic analysis of powdery mildew fungi is the availability of genomic DNA of suitable quality and in sufficient quantity. The so-called whole genome amplification technology has the potential to overcome this limitation. Here we present the application of phi29 DNA polymerase-mediated multiple displacement amplification (MDA) to amplify the whole genome of Podosphaera fusca, the main causal agent of powdery mildew in cucurbits, to address this problem. The genome coverage and fidelity of the MDA process was evaluated by PCR amplification and sequencing of two genetics markers: the nuclear rDNA internal transcribed spacer (ITS) regions and the mitochondrial cytochrome b gene (CYTB). Our results show that MDA is a valuable tool for molecular genetic analysis of powdery mildew fungi that can be used for a number of downstream applications in different fields, such as epidemiology and population genetics or systematics.
ISSN:0172-8083
1432-0983
DOI:10.1007/s00294-006-0117-7