Fluorescent markers of the microtubule cytoskeleton in Zymoseptoria tritici

Schuster, Martin; Kilaru, Sreedhar; Latz, M.; Steinberg, Gero

doi:10.1016/j.fgb.2015.03.005

Cited by 18 publications

(19 citation statements)

References 59 publications

(70 reference statements)

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“…However, comparative genomics among fungal species belonging to different taxa will allow identification of conserved key core determinants of hyphal tip growth. Furthermore, establishing molecular tools for cytological studies in nonmodel fungi, such as the wheat pathogen Z. tritici (166,(338)(339)(340)(341)(342)(343) or the grass pathogen species Ustilago bromivora and Brachypodium spp. (344), will enable more detailed comparison of the cytology of hyphal growth and fungal pathogenicity.…”

Section: Concluding Remarks and Future Directionsmentioning

confidence: 99%

Cell Biology of Hyphal Growth

et al. 2017

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Filamentous fungi are a large and ancient clade of microorganisms that occupy a broad range of ecological niches. The success of filamentous fungi is largely due to their elongate hypha, a chain of cells, separated from each other by septa. Hyphae grow by polarized exocytosis at the apex, which allows the fungus to overcome long distances and invade many substrates, including soils and host tissues. Hyphal tip growth is initiated by establishment of a growth site and the subsequent maintenance of the growth axis, with transport of growth supplies, including membranes and proteins, delivered by motors along the cytoskeleton to the hyphal apex. Among the enzymes delivered are cell wall synthases that are exocytosed for local synthesis of the extracellular cell wall. Exocytosis is opposed by endocytic uptake of soluble and membrane-bound material into the cell. The first intracellular compartment in the endocytic pathway is the early endosomes, which emerge to perform essential additional functions as spatial organizers of the hyphal cell. Individual compartments within septated hyphae can communicate with each other via septal pores, which allow passage of cytoplasm or organelles to help differentiation within the mycelium. This article introduces the reader to more detailed aspects of hyphal growth in fungi.

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Section: Concluding Remarks and Future Directionsmentioning

confidence: 99%

Cell Biology of Hyphal Growth

et al. 2017

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“…Here, we investigate the potential of MALCs to control fungal crop diseases. We predominantly use the Septoria blotch fungus Zymoseptoria tritici, which seriously threatens temperate-grown wheat 32 , and for which we established a raft of live-cell imaging marker strains e.g., 33,34 . We show that C 12 -G + targets fungal mitochondria and strongly inhibits ATP synthesis by reducing NADH oxidation and depolarizing the inner membrane.…”

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confidence: 99%

A lipophilic cation protects crops against fungal pathogens by multiple modes of action

et al. 2020

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The emerging resistance of crop pathogens to fungicides poses a challenge to food security and compels discovery of new antifungal compounds. Here, we show that mono-alkyl lipophilic cations (MALCs) inhibit oxidative phosphorylation by affecting NADH oxidation in the plant pathogens Zymoseptoria tritici, Ustilago maydis and Magnaporthe oryzae. One of these MALCs, consisting of a dimethylsulfonium moiety and a long alkyl chain (C18-SMe2+), also induces production of reactive oxygen species at the level of respiratory complex I, thus triggering fungal apoptosis. In addition, C18-SMe2+ activates innate plant defense. This multiple activity effectively protects cereals against Septoria tritici blotch and rice blast disease. C18-SMe2+ has low toxicity in Daphnia magna, and is not mutagenic or phytotoxic. Thus, MALCs hold potential as effective and non-toxic crop fungicides.

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“…To test for a role of the cytoskeleton in pore closure, we observed ZtHex1‐eGFP in laser‐injured cells that were treated with benomyl and latrunculin A. These inhibitors have been shown to disassemble microtubules and F‐actin in Z. tritici (Kilaru et al, ; Schuster, Kilaru, Latz, & Steinberg, ). Upon cell wounding, WBs moved into the septal pore in control cells, as well as in cells treated with benomyl and latrunculin A (Figure d).…”

Section: Resultsmentioning

confidence: 99%

“…It contains the gene for GFP, egfp , fused to gene Zthex1 , placed under the control of constitutive Zttub2 promoter and limited by the Zttub2 terminator (Kilaru, Schuster, Latz, et al, ). In detail, plasmid pCHex1eGFP carries a 12,530‐bp fragment of pCeGFPTub2 (Schuster et al, ; digested with Bsr GI), a 1149‐bp Z. tritici α‐tubulin promoter (amplified with SK‐Sep‐14 and SK‐Sep‐47; Table ), a 663‐bp full‐length Zthex1 gene without stop codon (amplified with SK‐Sep‐156 and SK‐Sep‐157; Table ), and a 717‐bp fragment, containing egfp (amplified with SK‐Sep‐16 and SK‐Sep‐78; Table ). The vector was generated by in vivo ligation of these DNA fragments in the yeast Saccharomyces cerevisiae (Kilaru & Steinberg, ).…”

Section: Methodsmentioning

confidence: 99%

ATP prevents Woronin bodies from sealing septal pores in unwounded cells of the fungus Zymoseptoria tritici

Steinberg

Schuster

Hacker

et al. 2017

Cellular Microbiology

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Septa of filamentous ascomycetes are perforated by septal pores that allow communication between individual hyphal compartments. Upon injury, septal pores are plugged rapidly by Woronin bodies (WBs), thereby preventing extensive cytoplasmic bleeding. The mechanism by which WBs translocate into the pore is not known, but it has been suggested that wound‐induced cytoplasmic bleeding “flushes” WBs into the septal opening. Alternatively, contraction of septum‐associated tethering proteins may pull WBs into the septal pore. Here, we investigate WB dynamics in the wheat pathogen Zymoseptoria tritici. Ultrastructural studies showed that 3.4 ± 0.2 WBs reside on each side of a septum and that single WBs of 128.5 ± 3.6 nm in diameter seal the septal pore (41 ± 1.5 nm). Live cell imaging of green fluorescent ZtHex1, a major protein in WBs, and the integral plasma membrane protein ZtSso1 confirms WB translocation into the septal pore. This was associated with the occasional formation of a plasma membrane “balloon,” extruding into the dead cell, suggesting that the plasma membrane rapidly seals the wounded septal pore wound. Minor amounts of fluorescent ZtHex1‐enhanced green fluorescent protein (eGFP) appeared associated with the “ballooning” plasma membrane, indicating that cytoplasmic ZtHex1‐eGFP is recruited to the extending plasma membrane. Surprisingly, in ~15% of all cases, WBs moved from the ruptured cell into the septal pore. This translocation against the cytoplasmic flow suggests that an active mechanism drives WB plugging. Indeed, treatment of unwounded and intact cells with the respiration inhibitor carbonyl cyanide m‐chlorophenyl hydrazone induced WB translocation into the pores. Moreover, carbonyl cyanide m‐chlorophenyl hydrazone treatment recruited cytoplasmic ZtHex1‐eGFP to the lateral plasma membrane of the cells. Thus, keeping the WBs out of the septal pores, in Z. tritici, is an ATP‐dependent process.

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Fluorescent markers of the microtubule cytoskeleton in Zymoseptoria tritici

Cited by 18 publications

References 59 publications

Cell Biology of Hyphal Growth

Cell Biology of Hyphal Growth

A lipophilic cation protects crops against fungal pathogens by multiple modes of action

ATP prevents Woronin bodies from sealing septal pores in unwounded cells of the fungus Zymoseptoria tritici

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