Editor: Bernard Paul
Development of a qPCR assay for specific quantification of Botrytis cinerea on grapes
Article first published online: 14 OCT 2010
© 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved
FEMS Microbiology Letters
Volume 313, Issue 1, pages 81–87, December 2010
How to Cite
Diguta, C. F., Rousseaux, S., Weidmann, S., Bretin, N., Vincent, B., Guilloux-Benatier, M. and Alexandre, H. (2010), Development of a qPCR assay for specific quantification of Botrytis cinerea on grapes. FEMS Microbiology Letters, 313: 81–87. doi: 10.1111/j.1574-6968.2010.02127.x
- Issue published online: 9 NOV 2010
- Article first published online: 14 OCT 2010
- Accepted manuscript online: 27 SEP 2010 12:00AM EST
- Received 5 July 2010; revised 16 September 2010; accepted 16 September 2010.Final version published online 14 October 2010.
- Botrytis cinerea;
The aim of this study was to develop a system for rapid and accurate real-time quantitative PCR (qPCR) identification and quantification of Botrytis cinerea, one of the major pathogens present on grapes. The intergenic spacer (IGS) region of the nuclear ribosomal DNA was used to specifically detect and quantify B. cinerea. A standard curve was established to quantify this fungus. The qPCR reaction was based on the simultaneous detection of a specific IGS sequence and also contained an internal amplification control to compensate for variations in DNA extraction and the various compounds from grapes that inhibit PCR. In these conditions, the assay had high efficiency (97%), and the limit of detection was estimated to be 6.3 pg DNA (corresponding to 540 spores). Our method was applied to assess the effects of various treatment strategies against Botrytis in the vineyard. Our qPCR assay proved to be rapid, selective and sensitive and may be used to monitor Botrytis infection in vineyards.