Genome sequences of many filamentous fungi are now available and additional genomes are currently being sequenced. One of the next strategic goals is to generate collections of tagged genes in order to establish a link between the several thousands of predicted genes and their function. Transposable elements have been invaluable for the identification and isolation of genes of interest as insertion of a transposon both disrupts and tags a gene. In an effort to exploit active transposons identified in the genome of Fusarium oxysporum as insertional mutagens, a binary system including the tagging element, MITE, and the transposase of a Tc1 element has been established as an efficient tool for gene-tagging in Fusarium graminearum. In this chapter, we provide an overview of the techniques used and highlight some of the critical steps for the application of this tool to other fungal species.
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BMC Res Notes
January 2025
Planta Piloto de Procesos Industriales Microbiológicos (PROIMI - CONICET), Tucumán, Argentina.
Background: Postharvest lemons are affected by several fungal infections, and as alternatives to chemical fungicides for combating these infections, different microbial biocontrol agents have been studied, with the Clavispora lusitaniae 146 strain standing out. Although strain 146 has proven to be an effective agent, the influence of a microbial biological control agent on the postharvest lemon microbiome has not been studied until now. Thus, this study aimed to evaluate how the epiphytic microbiome of postharvest lemons is affected by the application of the biocontrol yeast C.
View Article and Find Full Text PDFMycopathologia
January 2025
Teikyo University Institute of Medical Mycology (TIMM), 359 Otsuka, Hachioji, Tokyo, 192-0395, Japan.
We describe a novel Malassezia species named Malassezia polysorbatinonusus, isolated from a Japanese patient with seborrheic dermatitis. The internal transcribed spacer (ITS) region of the isolate (LSEM 4845) were only 94.7% identical to those of M.
View Article and Find Full Text PDFCell Res
January 2025
State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Sichuan Agricultural University, Chengdu, Sichuan, China.
Hydrogen peroxide (HO) is a ubiquitous signal regulating many biological processes, including innate immunity, in all eukaryotes. However, it remains largely unknown that how transcription factors directly sense HO in eukaryotes. Here, we report that rice basic/helix-loop-helix transcription factor bHLH25 directly senses HO to confer resistance to multiple diseases caused by fungi or bacteria.
View Article and Find Full Text PDFMethods Mol Biol
January 2025
Department of Immunobiology, University of Lausanne, Epalinges, Switzerland.
Fluorescence recovery after photobleaching (FRAP) can be employed to investigate membrane lipid mixing of vacuoles in live budding yeast cells and distinguish the fused, hemi-fused or non-fused states of these organelles under physiological conditions. Here, we describe a protocol for labeling the outer and inner leaflets of vacuoles in live cells that allow to detect hemifusion intermediates and, thus, identify components necessary for fusion pore opening.
View Article and Find Full Text PDFMethods Mol Biol
January 2025
School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju, South Korea.
Cell-free in vitro assays offer several advantages for elucidating molecular mechanisms underlying various biological processes. Here, we describe a simple and quantitative in vitro assay using isolated yeast microsomes to measure homotypic ER membrane fusion. In this assay, membrane fusion between ER microsomes is monitored by reconstitution of luciferase activity from split luciferase fragments.
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