Relationship between cell size and efficiency of synchronization during nitrogen-limited phased cultivation of Candida utilis.

Abstract

Under the phased method of cultivation the yeast Candida utilis grew and divided synchronously. The newly formed cells were relatively small, and a new cell cycle was not initiated until the cells could attain a certain minimum size (critical size). Although the cells expanded to some extent after division, the critical size was not reached until a fresh supply of medium was provided. With the arrival of the fresh supply of growth medium at the beginning of the phasing period, the cells expanded rapidly, and new cell cycles were initiated. The cells continued to expand until the growth-limiting nutrient (nitrogen source) was exhausted or until 90 min, which ever occurred first. Usually, buds emerged at a constant time after the start of the phasing period. The time of bud emergence was independent of the size attained by the cells during the expansion phase of growth. The results indicated that it was initiation of the cell cycle that was under size control, and not bud emergence. Bud emergence seemed to be under the control of a timer. The start of this timer seemed to be at or immediately after the beginning of the phasing period. Protein synthesis was essential for the initiation and expansion of buds. However, inhibition of protein synthesis by cycloheximide did not prevent unbudded cells or the parent portion of budded cells from expanding. Cycloheximide seemed to abolish the control mechanism(s) which prevented the cells from expanding after they had reached the maximum size.