Brodde, Laura
- Department of Forest Mycology and Plant Pathology, Swedish University of Agricultural Sciences
Research article2024Peer reviewedOpen access
Development of a molecular genetics and cell biology toolbox for the filamentous fungus Diplodia sapinea
Oostlander, Anne Geertje; Brodde, Laura; von Bargen, Miriam; Slippers, Bernard; Becker, Yvonne; Brandt, Ulrike; Klawonn, Frank; Grobler, Christiaan; Well, Lucas; Stenlid, Jan; Oliva, Jonas; Elfstrand, Malin; Fleissner, Andre
Abstract
Diplodia sapinea (Fr.) Fuckel is a widespread fungal pathogen affecting conifers worldwide. Infections can lead to severe symptoms, such as shoot blight, canker, tree death, or blue stain in harvested wood, especially in Pinus species. Its impact on forest health is currently intensified, likely due to climate change, posing an increasing threat to global ecosystems and forestry. Despite extensive and successful research on this pathogen system, fundamental questions about its biology and plant-associated lifestyle remain unanswered. Addressing these questions will necessitate the development of additional experimental tools, including protocols for molecular genetics and cell biology approaches. In this study, we continue to address this need by establishing an Agrobacterium-mediated genetic transformation protocol for D. sapinea, enabling targeted mutagenesis and heterologous gene expression. We utilized this methodology to localize the histone H2B by tagging it with the fluorescent protein mCherry. Additionally, we established a time- and space-efficient laboratory-scale infection assay using two-week-old Pinus sylvestris seedlings. Integrating these tools in a proof-of-concept study enabled the visualization of D. sapinea in planta growth through the fluorescently labeled reporter strain.
Published in
PLoS ONE
2024, volume: 19, number: 12, article number: e0308794
Publisher: PUBLIC LIBRARY SCIENCE
SLU Authors
Stenlid, Jan
- Department of Forest Mycology and Plant Pathology, Swedish University of Agricultural Sciences
- Department of Forest Mycology and Plant Pathology, Swedish University of Agricultural Sciences
Oliva Palau, Jonàs
- Department of Forest Mycology and Plant Pathology, Swedish University of Agricultural Sciences
- Department of Forest Mycology and Plant Pathology, Swedish University of Agricultural Sciences
Elfstrand, Malin
- Department of Forest Mycology and Plant Pathology, Swedish University of Agricultural Sciences
- Department of Forest Mycology and Plant Pathology, Swedish University of Agricultural Sciences
UKÄ Subject classification
Forest Science
Publication identifier
- DOI: https://doi.org/10.1371/journal.pone.0308794
Permanent link to this page (URI)
https://res.slu.se/id/publ/140136