Heterogeneity in establishment of polyethylene glycol-mediated plasmid transformations for five forest pathogenic Phytophthora species

• DOI: 10.1371/journal.pone.0306158
• Publication Date: 2024-09-10T17:23:28Z
• Authors: Erika N. Dort; Richard C. Hamelin
• Keywords: Phytophthora sojae; Phytophthora cactorum

Plasmid-mediated DNA transformation is a foundational molecular technique and the basis for most CRISPR-Cas9 gene editing systems. While plasmid transformations are well established many agricultural Phytophthora pathogens, development of this in forest Phytophthoras lacking. Given our long-term research objective to develop pathogenic species, we sought establish functionality polyethylene glycol (PEG)-mediated five species: P . cactorum , cinnamomi cryptogea ramorum syringae We used pathogen sojae species which PEG-mediated well-established, as control. Using protocol previously optimized tested with three different plasmids: two developed one fluorescent protein tagging. Out tested, successful transformation, indicated by stable growth transformants on high concentration antibiotic selective medium diagnostic PCR, was achieved only However, while were consistent stable, highly variable yielded very weak mycelial abnormal morphology. Our results indicate that best candidate move forward provide insight future optimization Phytophthoras.