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Open Access Methodology article

Transformation of Botrytis cinerea by direct hyphal blasting or by wound-mediated transformation of sclerotia

Shahar Ish - Shalom1, Aviva Gafni2, Amnon Lichter1 and Maggie Levy2*

Author Affiliations

1 Postharvest Science of Fresh Produce, The Volcani Center, ARO, Israel

2 The Department of Plant Pathology and Microbiology, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel

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BMC Microbiology 2011, 11:266  doi:10.1186/1471-2180-11-266

Published: 21 December 2011

Abstract

Background

Botrytis cinerea is a haploid necrotrophic ascomycete which is responsible for 'grey mold' disease in more than 200 plant species. Broad molecular research has been conducted on this pathogen in recent years, resulting in the sequencing of two strains, which has generated a wealth of information toward developing additional tools for molecular transcriptome, proteome and secretome investigations. Nonetheless, transformation protocols have remained a significant bottleneck for this pathogen, hindering functional analysis research in many labs.

Results

In this study, we tested three different transformation methods for B. cinerea: electroporation, air-pressure-mediated and sclerotium-mediated transformation. We demonstrate successful transformation with three different DNA constructs using both air-pressure- and sclerotium-mediated transformation.

Conclusions

These transformation methods, which are fast, simple and reproducible, can expedite functional gene analysis of B. cinerea.

Keywords:
Botrytis cinerea; sclerotium; Sclerotinia sclerotiorum; transformation; 'Bim-Lab'; electroporation