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Methods Mol Biol. 2011;712:129-35. doi: 10.1007/978-1-61737-998-7_11.

A straightforward protocol for electro-transformation of Phytophthora capsici zoospores.

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  • 1The Sainsbury Laboratory, John Innes Centre, Norwich, UK.

Abstract

Genome sequencing combined with high-throughput functional analyses has proved vital in our quest to understand oomycete-plant interactions. With the identification of effector molecules from Phytophthora spp. we can now embark on dissecting the mechanisms by which effectors modulate host processes and thus ensure parasite fitness. One of the key limitations, however, is to genetically modify Phytophthora and assess gene function during parasitism. Here, we describe a straightforward protocol that allows rapid transformation of Phytophthora capsici, an emerging model in oomycete biology. P. capsici is a broad host range pathogen that can infect a wide variety of plants under lab conditions making it a suitable model for detailed studies on oomycete-host interactions. This protocol relies on electroporation-assisted uptake of DNA in to motile zoospores and allows the rapid identification and characterization of genetically stable transformants.

PMID:
21359805
[PubMed - indexed for MEDLINE]
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