Fig. 5.

Mutation of milRNA2 blocks asexual development in response to injury. (a) Gel electrophoresis analysis of stem-loop RT-PCR products of the three most abundant milRNAs in the indicated RNAi mutants and two independent milRNA2 mutants. (b) Photographs show colonies of the indicated strains after 72 h of growth. (c) Time course of colony growth of the indicated strains. (b) and (c) Colonies were grown on PDA in darkness. Error bars represent ±sem. A one-way ANOVA test, followed by Tukey honest significant differences was used (**P <0.01). (d) The graph shows the percentage of hyphae that regenerate after injury for the indicated strains. 50 hyphae in total for each replicate were examined and three biological replicates (points) were performed per strain, lines indicate the average regeneration percentage. A one-way ANOVA test, followed by Tukey honest significant differences was used. Highlights in red indicate that there is a statistically significant difference when comparing the indicated strain with the WT (P <0.05). (e) Photographs show the conidiation phenotypes in response to injury of the WT strain, Δdcr2, ΔmilRNA2, and the dikaryon resulting from the fusion of Δdcr2/ΔmilRNAs. The experiment was performed on PDA and in complete darkness. (f) Expression level (reads per million) of the five milRNAs predicted in both the mutant and the dikaryons of milRNA2. (g) Length distribution of sRNA reads between 20 and 24 nt in the milRNA2 mutant, as well as the two dykarions.