An Improved Transformation System for Phytophthora cinnamomi Using Green Fluorescent Protein

Tingting Dai; Yue Xu; Xiao Yang; Xiao Yang; Binbin Jiao; Min Qiu; Junxin Xue; Felipe Arredondo; Felipe Arredondo; Brett M. Tyler; Brett M. Tyler

doi:10.3389/fmicb.2021.682754

Frontiers in Microbiology (Jul 2021)

An Improved Transformation System for Phytophthora cinnamomi Using Green Fluorescent Protein

Tingting Dai,
Yue Xu,
Xiao Yang,
Xiao Yang,
Binbin Jiao,
Min Qiu,
Junxin Xue,
Felipe Arredondo,
Felipe Arredondo,
Brett M. Tyler,
Brett M. Tyler

Affiliations

Tingting Dai: Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, China
Yue Xu: Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, China
Xiao Yang: Foreign Disease-Weed Science Research Unit, United States Department of Agriculture, Agricultural Research Service (ARS), Fort Detrick, MD, United States
Xiao Yang: ARS Research Participation Program, Oak Ridge Institute for Science and Education, Oak Ridge, TN, United States
Binbin Jiao: Technical Center for Animal, Plant and Food Inspection and Quarantine of Shanghai Customs, Shanghai, China
Min Qiu: Nanjing Agricultural University, Nanjing, China
Junxin Xue: Technical Center for Animal, Plant and Food Inspection and Quarantine of Shanghai Customs, Shanghai, China
Felipe Arredondo: Department of Botany and Plant Pathology, Center for Genome Research and Biocomputing, Oregon State University, Corvallis, OR, United States
Felipe Arredondo: Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR, United States
Brett M. Tyler: Department of Botany and Plant Pathology, Center for Genome Research and Biocomputing, Oregon State University, Corvallis, OR, United States
Brett M. Tyler: Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR, United States

DOI: https://doi.org/10.3389/fmicb.2021.682754
Journal volume & issue: Vol. 12

Abstract

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Phytophthora cinnamomi is a destructive pathogen causing root rot and dieback diseases on hundreds of economically and ecologically important plant species. Effective transformation systems enable modifications of candidate genes to understand the pathogenesis of P. cinnamomi. A previous study reported a polyethylene glycol and calcium dichloride (PEG/CaCl2)-mediated protoplast transformation method of P. cinnamomi. However, the virulence of the transformants was compromised. In this study, we selected ATCC 15400 as a suitable wild-type isolate for PEG/CaCl2 transformation using the green fluorescent protein after screening 11 P. cinnamomi isolates. Three transformants, namely, PcGFP-1, PcGFP-3, and PcGFP-5, consistently displayed a green fluorescence in their hyphae, chlamydospores, and sporangia. The randomly selected transformant PcGFP-1 was as virulent as the wild-type isolate in causing hypocotyl lesions on lupines. Fluorescent hyphae and haustoria were observed intracellularly and intercellularly in lupine tissues inoculated with PcGFP-1 zoospores. The potential application of this improved transformation system for functional genomics studies of P. cinnamomi is discussed.

Published in Frontiers in Microbiology

ISSN: 1664-302X (Online)
Publisher: Frontiers Media S.A.
Country of publisher: Switzerland
LCC subjects: Science: Microbiology
Website: http://www.frontiersin.org/journals/microbiology

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