Plant in vitro culture techniques are fundamental for research, propagation, and breeding. Automated phenotyping of in vitro cultures can revolutionize trait evaluation by transitioning to continuous and objective quantification, as well as by enhancing accuracy, speed, and efficiency. Limited research exists on automated sensor usage in plant tissue culture, mainly focusing on "plant-to-sensor" approaches. While reflection-based imaging techniques have dominated research to date, fluorescence-based imaging could offer advantages for the application of phenotyping in commercial in vitro propagation and plant research. We developed a new detector head for our “Phenomenon” plant phenotyping system to investigate the potential of fluorescence-based in situ monitoring of plant in vitro culture. In this study, we demonstrate the acquisition of fluorescence image data from plant in vitro cultures as an advanced imaging technique for phenotyping approaches. Over time and qualitatively, we were able to document the development of hairy roots in N. tabacum after transformation with Rhizobium rhizogenes carrying the recently developed reporter gene eYGFPuv.
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