Monitor in vivo the dynamic localization of Circadian Clock Components in Neurospora crassa
EMSL Project ID
60154
Abstract
Environmental controls on cellular metabolism, as exemplified by light and time of day, can modulate and even dwarf more commonly understood modes of cellular regulation. The expression of cellulases used in biofuel production, are often circadianly regulated, providing powerful and manipulable controls on the amount of cellulases that can be produced in fungi. In addition, it is likely that fungal-bacterial interactions are circadianly controlled in the mycorrhizosphere, in which it is hypothesized that trehalose produced by filamentous fungi is secreted in a circadian manner. Trehalose and other fungal metabolites stimulate taxis and interactions with soil bacteria that metabolize these compounds and in turn secrete antibacterials to the soil to ward off pathogenic bacteria that impact plant productivity. The filamentous fungus Neurospora crassa has been developed as a genetic model for the regulation and secretion of cellulases and metabolites, and it is also the principal fungal model system for the study of the effects of light and circadian clocks. Our goal is to visualize the spatiotemporal dynamics of clock proteins in the model system Neurospora. This information will contribute to understanding how the clock influences metabolic pathways and eventually how to enable the maximization of cellulase and metabolite secretion through the manipulation of the clock.We have invested considerable time in developing fluorescently tagged core circadian clock proteins through which the molecular events driving the circadian clock can be followed over the course of a day. However, because fungi in general, and Neurospora in particular, can grow at an extremely rapid speed (up to one micron per hour), specially designed microfluidic devices are needed to allow their growth to be followed. Arguably the best such device was the topic of a research paper (Geng et al., Compartmentalized microchannel array for high-throughput analysis of single cell polarized growth and dynamics) published by a team using EMSL and located at PNNL. Unfortunately no more of these devices exist at present, although the templates for them, and the expertise needed to reconstruct them, still do exist. The focus of this Limited Scope Proposal is to provide funds to facilitate the re-creation of several of these devices. Following their fabrication and our introduction to their use, these devices will be used to describe the cell biology surrounding the molecular events governing the creation of time information in Neurospora, as outlined in detail in the proposal.
Project Details
Project type
Limited Scope
Start Date
2022-07-18
End Date
2023-03-10
Status
Closed
Released Data Link
Team
Principal Investigator