Motility of fluorescent chitin and glucan synthase in hyphae of Z. tritici. (A) Domain structure of class V chitin synthases and β(1,3)-glucan synthase in Z. tritici (ZtChs5, Genbank accession number: SMQ47822.1; ZtGcs1, Genbank accession number: SMR60639.1), N. crassa (NcChs-5, Genbank accession number: XP_956333.2; NcGls1, Genbank accession number: XP_957980.1) and U. maydis (UmMcs1, Genbank accession number: XP_011389642.1; UmGcs1, Genbank accession number: XP_011387626.1). (B) Contrast inverted kymograph showing directed motility of eGFP3-ZtChs5. The direction relative to the hyphal tip is indicated by red arrow. Horizontal bar: 2 µm, vertical bar: 3 s. (C) Contrast inverted kymograph showing directed motility of eGFP4-ZtGcs1. The direction relative to the hyphal tip is indicated by red arrow. Horizontal bar: 2 µm, vertical bar: 3 s. (D) Bar chart showing average velocities of moving eGFP3-ZtChs5 and eGFP4-ZtGcs1 signals in hyphae of Z. tritici. Bars represent mean ± SEM, sample size n is indicated. Shapiro-Wilk testing revealed non-normal data distribution (P ≤ 0.05) and comparison used a nonparametric Mann-Whitney test, with n.s. indicating non-significant difference (P = 0.1099). (E) Bar chart show the flux of eGFP3-ZtChs5 and eGFP4-ZtGcs1 to the hyphal tip. Bars represent mean ± SEM, sample size n is indicated. Shapiro-Wilk testing revealed non-normal data distribution (P ≤ 0.05) and comparison used a nonparametric Mann-Whitney test, with n.s. indicating non-significant difference (P = 0.0840). (F) Bar chart show the flux of eGFP3-ZtChs5 and eGFP4-ZtGcs1 to the hyphal tip in the presence of the MT inhibitor benomyl. Bars represent mean ± SEM, sample size n is indicated. Shapiro-Wilk testing revealed non-normal data distribution (P ≤ 0.05) and comparison used a nonparametric Mann-Whitney test, with n.s. indicating non-significant difference (P = 0.5798). (G) Bar chart show the flux of eGFP3-ZtChs5 and eGFP4-ZtGcs1 to the hyphal tip in the presence of the F-actin inhibitor latrunculin A. Bars represent mean ± SEM, sample size n is indicated. Shapiro-Wilk testing revealed non-normal data distribution (P ≤ 0.05) and comparison used a nonparametric Mann-Whitney test, with n.s. indicating non-significant difference (P = 0.3869). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)