Abstract

The study of plant cell physiology is currently experiencing a profound transformation. Novel techniques allow dynamic in vivo imaging with subcellular resolution, covering a rapidly growing range of plant cell physiology. Several basic biological questions that have been inaccessible by the traditional combination of biochemical, physiological and cell biological approaches now see major progress. Instead of grinding up tissues, destroying their organisation, or describing cell- and tissue structure, without a measure for its function, novel imaging approaches can provide the critical link between localisation, function and dynamics. Thanks to a fast growing collection of available fluorescent protein variants and sensors, along with innovative new microscopy technologies and quantitative analysis tools, a wide range of plant biology can now be studied in vivo, including cell morphology & migration, protein localization, topology & movement, protein-protein interaction, organelle dynamics, as well as ion, ROS & redox dynamics. Within the cell, genetic targeting of fluorescent protein probes to different organelles and subcellular locations has started to reveal the stringently compartmentalized nature of cell physiology and its sophisticated spatiotemporal regulation in response to environmental stimuli. Most importantly, such cellular processes can be monitored in their natural 3D context, even in complex tissues and organs – a condition not easily met in studies on mammalian cells. Recent new insights into plant cell physiology by functional imaging have been largely driven by technological developments, such as the design of novel sensors, innovative microscopy & imaging techniques and the quantitative analysis of complex image data. Rapid further advances are expected which will require close interdisciplinary interaction of plant biologists with chemists, physicists, mathematicians and computer scientists. High-throughput approaches will become increasingly important, to fill genomic data with ‘life’ on the scale of cell physiology. If the vast body of information generated in the -omics era is to generate actual mechanistic understanding of how the live plant cell works, functional imaging has enormous potential to adopt the role of a versatile standard tool across plant biology and crop breeding. We welcome original research papers, methodological papers, reviews and mini reviews, with particular attention to contributions in which novel imaging techniques enhance our understanding of plant cell physiology and permits to answer questions that cannot be easily addressed with other techniques.

Highlights

  • The last quarter of a century has brought major developments in the acquisition of images from plants through improvements in microscopy equipment, software and technique

  • Förster resonance energy transfer (FRET) and how sensitized emission may be used for quantification in vivo imaging is reviewed by Müller et al (2013) who discuss a set of methods that allows for the analysis of molecular interactions, in the light of recent developments in fluorescence microscopy, which have achieved higher spatial and temporal resolution as well as a much-improved sensitivity

  • A comprehensive overview of FRET imaging is given with a focus on fluorescent proteins and the procedure and analysis of sensitized emission, which allows for a fast and repetitive monitoring of FRET efficiencies as required for the investigation of dynamic plant cells

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Summary

Introduction

The last quarter of a century has brought major developments in the acquisition of images from plants through improvements in microscopy equipment, software and technique. The articles contributed to this research topic are indicative of the work emerging from the plant imaging community and cover, variously, the characterization of individual protein functions; localization and interactions; the use of physiological biosensors; spectroscopic techniques, which utilize autofluorescence of plant tissues and label-free approaches; developmental studies and software engineering.

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