Thermal imaging as a tool for studying circadian rhythms in roots

Plants rely on the circadian clock to anticipate daily environmental fluctuations and to coordinate key physiological, metabolic, and developmental processes. Most if not all plant cells have semi-autonomous circadian oscillators. Roots possess a modified yet robust circadian oscillator that is entrained by external cues such as light and temperature to synchronize nutrient uptake, water transport, and metabolic activity. It has been shown that the root and shoot oscillators can communicate through long-distance signals including mobile proteins and carbon assimilates such as sucrose. Moreover, recent studies indicate root-microbe interactions; root-associated microbial communities exhibit diurnal oscillations structured by the host circadian system, while microbes can in turn modulate the circadian period and rhythmic outputs of the plant. However, in general, while the shoot circadian oscillator has been extensively characterized, much less is known about the root circadian system. Progress has been hampered by a lack of high-throughput, non-invasive methods to study root rhythmicity. Existing methods including luciferase reporters, quantitative RT-PCR, and microscopy remain limited by cost, destructive sampling, or require transgenic lines with reporter genes. We have developed a thermal infrared imaging platform that enables non-invasive, high resolution of circadian rhythms in roots across plant species and growth conditions. We show that our system can be used to analyse metabolite and microbial effects on root circadian regulation. This platform provides new opportunities to investigate below-ground circadian regulation and the possibilities of harnessing the root clock to enhance plant performance and resilience.