A method for the synchronous induction of large numbers of telotrochs in Vorticella convallaria by monocalcium phosphate at low pH

We have devised a two step method for the synchronous induction of telotrochs in the peritrich ciliate, Vorticella convallaria. The method is easy, reliable, and allows us to study the earliest events of telotroch formation at the ultrastructural, biochemical, and molecular levels. The steps involved are: (1) excising the cell body from the stalk in a large population (7.4 X 10⁴ cells) of EDTA-treated, attached cells by the application of monocalcium phosphate monohydrate solution at pH 3.2, (2) rinsing and suspending the isolated cell bodies in inorganic medium. Within 90 min, 80% of the population forms telotrochs. Analysis of factors that are important for maximum stalk excision and transformation shows that the population must not be older than 2 d and the most effective concentration of monocalcium phosphate is 4.8 mM for a 20 min exposure. The most effective monocalcium phosphate is in the monohydrated form. A pH value of 3.2, produced by the addition of hydrochloric acid in the presence or absence of calcium is not sufficient to initiate stalk excision and telotroch formation. This observation leads us to conclude that stalk excision is dependent on monocalcium phosphate or its hydrolysis products.