Journal Article

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

Levy, Maggie, Ish - Shalom, Shahar, Lichter, Amnon, Gafni, Aviva

2011

Overview

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 .

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Analysis / Applied microbiology / Ascomycetes / Bim-Lab / Biological Microscopy / Biomedical and Life Sciences / Biotechnology - methods / Botrytis / Botrytis - genetics / Botrytis cinerea / Deoxyribonucleic acid / DNA / DNA, Fungal - analysis / Efficiency / Electroporation / Gene expression / Gene Transfer Techniques / Genetic aspects / Genetic transformation / Genomes / Grey mold / Hyphae - genetics / Life Sciences / Methodology / Methodology Article / Methods / Microbiology / Mycology / Parasitology / Pathogens / Physiological aspects / Plant diseases / Plant species / R&D / Research & development / Sclerotia / Sclerotinia sclerotiorum / sclerotium / secretome / Transformation / Transformation, Genetic / Virology