RaPiD-chamber: Easy to self-assemble live-imaging chamber with adjustable LEDs allows to track small differences in dynamic plant movement adaptation on tissue level

Abstract

Plants rely on fine-tuning organ movement to ensure their survival and productivity. Even subtle loss of directional growth orchestration can result in a huge impact when the plant is impaired to adapt to everchanging environment, where it is exposed to manifold exogenous stimuli simultaneously. We present a newly designed chamber to obtain live images to track differences of organ growth and movement, called RaspberyPi Dark Chambers (RaPiD-chamber). The RaPiD-chamber is easy to self-assemble and cost efficient, and allows to monitor continuous growth of etiolated seedlings, as well their response to light of different wavelength and from chosen positions. We tested the advice by comparing hypocotyl elongation rate and response to unilateral white and blue light exposure of Arabidopsis thaliana Col-0. Additionally, we compared the elongation rate of etiolated hypocotyls between Col0 and kin10, a mutant lacking the catalytic subunit of the cellular signaling hub SUCROSE NON-FERMENTING RELATED KINASE 1 (SnRK1). kin10 is known for its diminished ability to control hypocotyl elongation. We compared the growth dynamics of etiolated Col0 versus kin10 as a case study. Without further energy source supplementation to the growth medium the mutant cannot keep up with hypocotyl elongation. Additionally, continuous observation of the dark grown seedlings allowed us to determine a shift in the dynamics of apical hook angle formation for the mutant.

Dynamic Dark Root Chamber: Advancing non-invasive phenotyping of roots kept in darkness using infrared imaging

“…IR LEDs seem to have no obvious effect on root fitness (Wells et al, 2012;Shi et al, 2018;Dermendjiev et al, 2023). We were previously using IR LEDs to study growth dynamics of etiolated seedlings and we couldn't distinguish growth behaviour from control plants in fully etiolated growth conditions (Kashkan, Garcia-González, et al, 2022;Kashkan, Hrtyan, et al, 2022). This approach can be used to monitor root outgrowth from germination, on agar plates, germination paper or soil, and to study various dynamic root traits and responses.…”

“…If the material tends to heat up, we recommend adding fans to cool down the chamber. In this manuscript, we use solid cardboard for the corpus of the chamber, because it is inexpensive, easy to manipulate, and proven to be effective in previous imaging setups (Kashkan, Garcia-González, et al, 2022;Kashkan, Hrtyan, et al, 2022).…”

Recent insights into metabolic and signalling events of directional root growth regulation and its implications for sustainable crop production systems

“…TOR activity inhibition is enhanced when environmental signals are additionally perceived and transmitted by SnRK2-dependent pathways ( Figure 2B ) ( Pu et al., 2017 ; Huang et al., 2019 ; Henriques et al., 2022 ). The complex interactions of SnRK1, SnRK2, TOR and autophagy that maintain cellular function under different growth conditions, which underpins plant growth and fitness, are currently intensively studied and will provide a better understanding of the environmental impact on plant productivity ( Hill et al., 2013 ; Barrada et al., 2015 ; Li et al., 2018 ; Ghatak et al., 2022a ; Kashkan et al., 2022 ).…”