Demonstration of RNA interference by bacterial feeding in Stentor polymorphus

Abstract

Stentor is a genus of large trumpet-shaped unicellular organisms in the ciliate phylum. Classically they have been used as models of cellular morphogenesis due to their large size and ability to regenerate, but some Stentor species have features that make them useful models for other types of studies as well. Stentor polymorphus is a widely distributed species that harbors green algal endosymbionts from the Chlorella genus. While interesting phenomenology in this species has been described, molecular tools have never been developed in this system. As technology has advanced, the use of emerging models like S. polymorphus has become more prevalent, and recently a set of transcriptomes for S. polymorphus was published. However, there are still technical hurdles to using S. polymorphus as an effective experimental system in the lab. Here I describe the identification and culture of a S. polymorphus population from North Carolina as well as the identification and cloning of homologs of -tubulin and the morphogenesis gene mob-1. Additionally, I demonstrate that RNA interference (RNAi) by feeding is effective against both of these homologs in S. polymorphus. The phenotypes observed in S. polymorphus were similar to phenotypes previously validated in S. coeruleus, a related Stentor species. A direct comparison of feeding RNAi between the two species revealed that RNAi appeared to be less effective in S. polymorphus. The ability to perform RNAi in S. polymorphus strengthens its use as an emerging model for exploring mechanisms of unicellular morphogenesis and regeneration or host-symbiont interactions and suggests that RNAi by bacterial feeding might be more broadly effective across the Stentor genus.