Nuclear dynamics in the homothallic ascomycete Sordaria fimicola

Krobanan, Kulsumpun; Shen, Wei-Chiang

doi:10.1186/s40529-018-0233-y

Cited by 3 publications

(9 citation statements)

References 26 publications

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“…Previous studies reported that light responses in S. fimicola include carotenoid accumulation and phototropism of the perithecial beak. Recently, a gene delivery system and cytological fluorescence study in S. fimicola was reported (44). The goal of this work was to gain a fundamental understanding of blue-light signal transduction in S. fimicola for sexual development, carotenoid biosynthesis, and phototropism.…”

Section: Discussionmentioning

confidence: 99%

“…Nucleic acid manipulation and analysis. For PCR screening and Southern blot analysis, genomic DNA (gDNA) of S. fimicola was extracted by using modified cetyltrimethylammonium bromide (CTAB) method as previously described (44). Briefly, Sordaria strains were grown on cellophane-covered malt extract agar I medium, and mycelia grown on cellophane were peeled and dipped into liquid nitrogen.…”

Section: Methodsmentioning

confidence: 99%

“…Next, a 1.9-kb 3= fragment downstream of the Sfwc-1 LOV domain was amplified with WC1338/WC1436, purified, cut with XhoI and PstI, and subcloned into pCAMBIA::5=Sfwc-1-hyg to generate pCAMBIA::5=-3=Sfwc-1 ⌬LOV -hyg. The disruption plasmid construct was electroporated into Agrobacterium strains and Agrobacterium containing a vector was used for Agrobacterium-mediated protoplast transformation as previously described (44). To identify putative Sfwc-1 (⌬lov) mutants, the primer pairs WC1398/WC1320 (5= flanking the LOV domain) and WC885/WC1436 (3= flanking the LOV domain) were used for screening of gene replacement mutants.…”

Section: Methodsmentioning

confidence: 99%

“…The plasmid was transformed back into Sfwc-1 (⌬lov) mutant strains. Transformation was performed as previously described (44). To screen for the complementation strains, the primer pairs WC1398/WC1337 and WC1688/WC1689 in the deleted LOV domain region and neo cassette, respectively, were used for PCR screening.…”

Section: Methodsmentioning

confidence: 99%

“…Collectively, these reports have provided information about the effects of light on the life cycle of S. fimicola from the vegetative to reproductive stage, although phenotypebased studies were predominantly emphasized (43). More recently, a gene delivery system and fluorescence microscopy studies have been reported for S. fimicola (44).…”

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confidence: 99%

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The Blue-Light Photoreceptor Sfwc-1 Gene Regulates the Phototropic Response and Fruiting-Body Development in the Homothallic Ascomycete Sordaria fimicola

Krobanan

Liang²,

Chiu³

et al. 2019

Appl Environ Microbiol

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Sordaria fimicola, a coprophilous ascomycete, is a homothallic fungus that can undergo sexual differentiation with cellular and morphological changes followed by multicellular tissue development to complete its sexual cycle. In this study, we identified and characterized the blue-light photoreceptor gene in S. fimicola. The S. fimicola white collar-1 photoreceptor (SfWC-1) contains light-oxygen-voltage-sensing (LOV), Per-Arnt-Sim (PAS), and other conserved domains and is homologous to the WC-1 blue-light photoreceptor of Neurospora crassa. The LOV domain of Sfwc-1 was deleted by homologous recombination using Agrobacterium-mediated protoplast transformation. The Sfwc-1(Δlov) mutant showed normal vegetative growth but produced less carotenoid pigment under illumination. The mutant showed delayed and less-pronounced fruiting-body formation, was defective in phototropism of the perithecial beaks, and lacked the fruiting-body zonation pattern compared with the wild type under the illumination condition. Gene expression analyses supported the light-induced functions of the Sfwc-1 gene in the physiology and developmental process of perithecial formation in S. fimicola. Moreover, green fluorescent protein (GFP)-tagged SfWC-1 fluorescence signals were transiently strong upon light induction and prominently located inside the nuclei of living hyphae. Our studies focused on the putative blue-light photoreceptor in a model ascomycete and contribute to a better understanding of the photoregulatory functions and networks mediated by the evolutionarily conserved blue-light photoreceptors across diverse fungal phyla. IMPORTANCE Sordaria sp. has been a model for study of fruiting-body differentiation in fungi. Several environmental factors, including light, affect cellular and morphological changes during multicellular tissue development. Here, we created a light-oxygen-voltage-sensing (LOV) domain-deleted Sfwc-1 mutant to study blue-light photoresponses in Sordaria fimicola. Phototropism and rhythmic zonation of perithecia were defective in the Sfwc-1(Δlov) mutant. Moreover, fruiting-body development in the mutant was reduced and also significantly delayed. Gene expression analysis and subcellular localization study further revealed the light-induced differential gene expression and cellular responses upon light stimulation in S. fimicola.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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The Blue-Light Photoreceptor Sfwc-1 Gene Regulates the Phototropic Response and Fruiting-Body Development in the Homothallic Ascomycete Sordaria fimicola

Krobanan

Liang²,

Chiu³

et al. 2019

Appl Environ Microbiol

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Nucleus-directed fluorescent reporter system for promoter studies in the ectomycorrhizal fungus Laccaria bicolor

Kemppainen

Pardo

2021

Journal of Microbiological Methods

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Analysis of Nuclear Dynamics in Nematode-Trapping Fungi Based on Fluorescent Protein Labeling

Zhou,

He,

Zhang

et al. 2023

JoF

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Nematophagous fungi constitute a category of fungi that exhibit parasitic behavior by capturing, colonizing, and poisoning nematodes, which are critical factors in controlling nematode populations in nature, and provide important research materials for biological control. Arthrobotrys oligospora serves as a model strain among nematophagous fungi, which begins its life as conidia, and then its hyphae produce traps to capture nematodes, completing its lifestyle switch from saprophytic to parasitic. There have been many descriptions of the morphological characteristics of A. oligospora lifestyle changes, but there have been no reports on the nuclear dynamics in this species. In this work, we constructed A. oligospora strains labeled with histone H2B–EGFP and observed the nuclear dynamics from conidia germination and hyphal extension to trap formation. We conducted real-time imaging observations on live cells of germinating and extending hyphae and found that the nucleus was located near the tip. It is interesting that the migration rate of this type of cell nucleus is very fast, and we speculate that this may be related to the morphological changes involved in the transformation to a predatory lifestyle. We suggest that alterations in nuclear shape and fixation imply the immediate disruption of the interaction with cytoskeletal mechanisms during nuclear migration. In conclusion, these findings suggest that the signal initiating nuclear migration into fungal traps is generated at the onset of nucleus entry into a trap cell. Our work provides a reference for analysis of the dynamics of nucleus distribution and a means to visualize protein localization and interactions in A. oligospora.

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Nuclear dynamics in the homothallic ascomycete Sordaria fimicola

Cited by 3 publications

References 26 publications

The Blue-Light Photoreceptor Sfwc-1 Gene Regulates the Phototropic Response and Fruiting-Body Development in the Homothallic Ascomycete Sordaria fimicola

The Blue-Light Photoreceptor Sfwc-1 Gene Regulates the Phototropic Response and Fruiting-Body Development in the Homothallic Ascomycete Sordaria fimicola

Nucleus-directed fluorescent reporter system for promoter studies in the ectomycorrhizal fungus Laccaria bicolor

Analysis of Nuclear Dynamics in Nematode-Trapping Fungi Based on Fluorescent Protein Labeling

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