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Publication type: Conference poster
Type of review: Not specified
Title: Simultaneous detection and identification of pathogenic fungi in wheat using a DNA macroarray
Authors: Palmisano, Marilena
Wohler, Christian
Boos, Jürg
Kuhn, Roger
Sierotzki, H.
Bolsinger, M.
DOI: 10.21256/zhaw-4291
Proceedings: Book of Abstracts: 2nd Symposium on Horticulture in Europe - SHE 2012
Page(s): 171
Conference details: 2nd Symposium on Horticulture in Europe, Angers, 1-5 July 2012
Issue Date: 2012
Publisher / Ed. Institution: ZHAW Zürcher Hochschule für Angewandte Wissenschaften
Language: English
Subjects: IUNR; SPEZI; ZHO
Subject (DDC): 572: Biochemistry
580: Plants (Botany)
Abstract: The detection of economically important pathogens is a key element in sustainable wheat production and a prerequisite for crop protection. The objective of the project was to develop a DNA macroarray for fast and cost-effective detection of nine pathogenic fungi in wheat: Fusarium graminearum, Fusarium culmorum, Fusarium poae, Microdochium nivale var. majus, Microdochium nivale var. nivale, Puccinia recondita, Septoria tritici, Septoria nodorum and Pyrenophora tritici-repentis. Methodically, a macroarray is similar to a microarray but without the need for expensive equipment. PCR labelled samples of DNA are hybridized to pathogen-specific oligonucleotides (probes) anchored to a solid support. A positive reaction between an amplicon and a perfectly matched oligonucleotide generates a chemiluminescent signal which can be detected by a plate reader. The macroarray is sensitive enough to detect single nucleotide polymorphism (SNPs). Sample analysis is simple, fast, cost-effective, fully automated and suitable for high throughput screening. In this project, the nine wheat pathogens were detected within 6 hours simultaneously in a single sample using between one to four different species-specific probes for each pathogen. Species-specific detector oligonucleotides were designed based on the β-tubulin and/or succinate dehydrogenase region of fungal DNA. The detection limit of the DNA macroarray technique particularly depends on the pathogen-specific oligonucleotides deployed. The necessity for monitoring pathogenic fungi in wheat production and for prediction of crop yield has been recognized for a long time. The DNA macroarray responds very sensitively and has the potential to recognize pathogenic fungi earlier with reference to the cultivation period than a conventional PCR. This means that the DNA macroarray can detect genomic DNA from fungi in a lower potency than the conventional PCR. One benefit of the DNA macroarray for detection of fungal pathogens in wheat is its increased specificity and the other its application to a large number of microorganisms which can be detected in a single assay. This technology has been proven to be relatively cost-effective compared with real-time PCR or microarrays. This project was financially supported by the Commission of Technology and Innovation CTI in Berne, Switzerland.
Fulltext version: Published version
License (according to publishing contract): Licence according to publishing contract
Departement: Life Sciences and Facility Management
Organisational Unit: Institute of Natural Resource Sciences (IUNR)
Appears in collections:Publikationen Life Sciences und Facility Management

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