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Establishment of a simple and efficient agrobacterium-mediated transformation system for Phytophthora palmivora D. WU (1), N. Navet (1), M. Tian (1) (1) University of Hawaii at Manoa, U.S.A.
The genus Phytophthora contains a large number of destructive plant pathogens that severely threaten agricultural production and natural ecosystems. Among them is the broad host range pathogen P. palmivora, which infects many economically important hosts such as papaya, cacao and rubber tree. An essential way to dissect their pathogenesis mechanisms is genetic modification of candidate genes, which requires effective transformation systems. Comm used Phytophthora transformation systems include PEG/CaCl2 mediated protoplast transformation and zoospore electroporation, both of which often lead to multi-copy integration in the genome. Although a higher copy number may be associated with higher level of gene expression and gene silencing, multi-copy integration randomly disrupts multiple genes varying among transformants and therefore complicates gene function analyses. Agrobacterium-mediated transformation (AMT) circumvents this issue as it mainly generates single-copy insertion. An AMT method previously reported for P. infestans and P. palmivora has barely been used in oomycete research due to low success and low reproducibility of generating transformants. We largely modified the protocol to make it a simple, efficient and highly reproducible approach to transform P. palmivora, which will be presented in the meeting. This approach will greatly facilitate P. palmivora functional genomic studies, and is expected to be adaptable for transforming other Phytophthora spp.
Abstract Number:
P9-312 Session Type:
Poster
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