Scanning and negative-staining electron microscopy of protoplast regeneration of a wild-type and two chitin synthase mutants in the pathogenic yeast Candida glabrata

• DOI: 10.1093/jmicro/dfq082
• Publication Date: 2011-01-08T01:13:41Z
• Authors: Y. Namiki; K. Ueno; H. Mitani; E. V. Virtudazo; M. Ohkusu; K. Shimizu; S. Kawamoto; H. Chibana; M. Yamaguchi
• Keywords: Protoplast; Chitin synthase; Negative stain; Strain (injury); Wild type

Protoplast regeneration of a wild-type and two mutant strains Candida glabrata defective in CHS3 homologues encoding class IV chitin synthase Saccharomyces cerevisiae was examined by scanning negative-staining electron microscopy. In the strain, small particles short filaments appeared on protoplast surface at 10 min, filamentous materials covered entire 1 h, granular started filling interspaces 2 h completed 6 h. The consisted microfibrils various widths ranging from ≤5 to 40 nm, composed β-glucan. Protoplasts Δchs3A Δchs3B essentially same process as although it took more time. These results suggest that CHS3A CHS3B genes may have important roles cell wall formation during regeneration, but can be compensated other enzymes.