Logo image
In situ PCR for detection and identification of fungal species
Journal article   Open access   Peer reviewed

In situ PCR for detection and identification of fungal species

L. Bindslev, R.P. Oliver and B. Johansen
Mycological Research, Vol.106(3), pp.277-279
2002
pdf
in_situ_PCR.pdfDownloadView
Published (Version of Record) Open Access
url
Link to Published Version *Subscription may be requiredView

Abstract

PCR and DNA sequence analysis have become standard tools for identification, detection and phylogenetic analysis of fungi. A large number of species are incapable of growth in the laboratory, making the preparation of pure DNA problematical. The amplification of DNA samples from impure material is subject to misinterpretation if more than one species is present. To overcome this problem, we designed an in situ PCR technique that links PCR amplification to the light microscopic image. The amplified tissue is stained, thus confirming which morphotype has been amplified. The PCR product can then be sequenced. We tested the technique on fixed Blumeria graminis spores and mycelia using primers derived from the sequence of the gene encoding the catalytic subunit of protein kinase A (bka1). This is the first report of in situ PCR on phytopathogenic fungal material. This technique allows positive confirmation of the origin of genes cloned from obligate pathogenic fungi and could be adapted for use on any samples containing mixed fungal species.

Details

UN Sustainable Development Goals (SDGs)

This output has contributed to the advancement of the following goals:

#3 Good Health and Well-Being

Source: InCites

Metrics

997 File views/ downloads
166 Record Views

InCites Highlights

These are selected metrics from InCites Benchmarking & Analytics tool, related to this output

Collaboration types
Domestic collaboration
International collaboration
Citation topics
1 Clinical & Life Sciences
1.54 Molecular & Cell Biology - Genetics
1.54.1454 PCR Techniques
Web Of Science research areas
Mycology
ESI research areas
Plant & Animal Science
Logo image