Class V chitin synthase and β(1,3)-glucan synthase co-travel in the same vesicle in Zymoseptoria tritici

•Native chitin (Chs5) and glucan synthase (Gsc1) visualised in the pathogen Zymoseptoria tritici.•Chs5 and Gsc1 are transported along microtubules.•Chs5 and Gsc1 do localise to the apical plasma membrane, but not the Spitzenkörper.•Light and electron microscopy how co-travel of Chs5 and Gsc1 in the same secretory vesicle.•Enzyme delivery in Z. tritici is different from Neurospora crassa, but similar to Ustilago maydis. The fungal cell wall consists of proteins and polysaccharides, formed by the co-ordinated activity of enzymes, such as chitin or glucan synthases. These enzymes are delivered via secretory vesicles to the hyphal tip. In the ascomycete Neurospora crassa, chitin synthases and β(1,3)-glucan synthase are transported in different vesicles, whereas they co-travel along microtubules in the basidiomycete Ustilago maydis. This suggests fundamental differences in wall synthesis between taxa. Here, we visualize the class V chitin synthase ZtChs5 and the β(1,3)-glucan synthase ZtGcs1 in the ascomycete Zymoseptoria tritici. Live cell imaging demonstrate that both enzymes co-locate to the apical plasma membrane, but are not concentrated in the Spitzenkörper. Delivery involves co-transport along microtubules of the chitin and glucan synthase. Live cell imaging and electron microscopy suggest that both cell wall synthases locate in the same vesicle. Thus, microtubule-dependent co-delivery of cell wall synthases in the same vesicle is found in asco- and basidiomycetes.