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Quantification of Symplasmic Phloem Loading Capacity with Live-Cell Microscopy.

Helle Juel Martens¹, Chen Gao², Johannes Liesche³

¹Department of Geosciences and Natural Resources, University of Copenhagen, Copenhagen, Denmark.

Methods in Molecular Biology (Clifton, N.J.)

|June 15, 2019

Summary

This study quantifies sugar transport in plants using fluorescent tracers. The method measures diffusion through plasmodesmata, revealing how sugars move from leaves to other organs via phloem vascular tissue.

Keywords:

Carbon allocation Fluorescent tracer Live-cell microscopy Phloem loading Photobleaching Plasmodesmata

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Area of Science:

Plant Biology
Molecular Plant Physiology
Plant Vascular Transport

Background:

Photosynthesis produces sugars in leaves, which are then transported to other plant organs via the phloem vascular tissue.
Three primary mechanisms for phloem loading exist, differing in active transport requirements and passive intercellular diffusion via plasmodesmata.
Quantifying diffusion through plasmodesmata is crucial for understanding sugar allocation in plants.

Purpose of the Study:

To describe a simple method for quantifying plasmodesmata-mediated intercellular diffusion.
To assess diffusion across the mesophyll-bundle sheath and bundle sheath-phloem cell interfaces.
To provide insights into sugar transport mechanisms in plants.

Main Methods:

Utilized live-cell microscopy to observe the movement of fluorescent tracers.
Employed carboxyfluorescein as a fluorescent tracer loaded into intact leaves.
Monitored tracer diffusion using confocal microscopy following photobleaching of bundle sheath cells.

Main Results:

Successfully quantified plasmodesmata-mediated intercellular diffusion across key plant tissue interfaces.
Demonstrated the movement of fluorescent tracers as a proxy for sugar molecule transport.
Provided a method to differentiate between active and passive transport mechanisms in phloem loading.

Conclusions:

The described method allows for the quantification of diffusion across the mesophyll-bundle sheath and bundle sheath-phloem cell interfaces.
This technique offers a valuable tool for studying sugar transport and phloem loading mechanisms in plants.
Understanding these transport dynamics is essential for plant physiology and crop science.