Non-invasive imaging of leaf area using a mobile phone in the EcoFAB-N v1

This protocol describes the non-invasive phenotyping steps in the EcoFAB-N chamber published in the study Kuang et al., "N-dependent dynamics of root growth and nitrate and ammonium uptake are altered by bacteria Herbaspirillum seropedicae in cereal model Brachypodium distachyon" Figure legends (attached images): Fig. SM2. EcoFAB-N imaged from thee sides using the Plant screen mobile application on a mobile phone and analyzed with Color segmentation software. (a) EcoFAB-N on a grid attached to plastic plate on clean bench. Grid of known size (here, 20 mm x 20 mm squares, both black and white grid background work, black allows better visualization of roots, white is sufficient for leaves) (b) Shoot images (leaves) taken with a mobile phone positioned at three ways (1, 2, 3) with Plant screen mobile (PSM) software (Müller-Linow et al., 2019). Image analysis process with Color segmentation software guide. Steps for single (c) and multiple images analysis (d). Fig. SM3 Plant screen mobile software (PSM) guide. (a) PSM App icon. (b-f) PSM settings. Relevant references to the original EcoFAB chamber and the imaging software: Gao J, Sasse J, Lewald KM, Zhalnina K, Cornmesser LT, Duncombe TA, Yoshikuni Y, Vogel JP, Firestone MK, Northen TR. 2018. Ecosystem Fabrication (EcoFAB) Protocols for The Construction of Laboratory Ecosystems Designed to Study Plant-microbe Interactions. J Vis Exp(134). Müller-Linow M, Pinto-Espinosa F, Scharr H, Rascher U. 2015. The leaf angle distribution of natural plant populations: assessing the canopy with a novel software tool. Plant Methods11(1): 11. Müller-Linow M, Wilhelm J, Briese C, Wojciechowski T, Schurr U, Fiorani F. 2019.Plant Screen Mobile: an open-source mobile device app for plant trait analysis. Plant Methods15(1): 2. Sasse J, Kant J, Cole BJ, Klein AP, Arsova B, Schlaepfer P, Gao J, Lewald K, Zhalnina K, Kosina S, et al.,. 2019.Multilab EcoFAB study shows highly reproducible physiology and depletion of soil metabolites by a model grass. The New phytologist 222(2): 1149-1160.