Fig 2. Budding pattern of elm1Δ fus3Δ cells.

All of indicated strains (MY8092, 12158, 12886 and 12948) were BY4741-derivative haploids and grown to exponential phase in liquid YPD at 30°C. S288C-derivative diploid (MY13411) was used as a positive control of bipolar budding. (A) Budding pattern was examined by time lapse microscopy as described in Materials and methods. Representative images are shown at 10-min interval. Arrows and arrow heads indicate the daughters that emerge adjacent or opposite to the preceding division site, respectively. (B) Cells were classified by number and distribution of bud scars. For each sample, more than 200 cells were counted and the score was expressed as a percentage. N.D, not determined. (C and D) Cells were stained with 0.1% calcofluor. The classified budding patterns are denoted as numbers that correspond to that of (B). (C) Arrows indicate birth scar. (D) Representative cluster of elm1Δ fus3Δ cells is shown with an interpretative drawing of the budding pattern (right). Arrow head (left) and black ring (drawing) indicate a bud scar. Arrows indicate abnormal chitin accumulations. (A, C, and D) Bar, 10 μm.