Chapter 8
In Vivo Imaging of Mobile mRNAs in Plants Using MS2
Kai-Ren Luo, Nien-Chen Huang, and Tien-Shin Yu
Abstract
Multicellular organisms rely on systemic signals to orchestrate diverse developmental and physiological
programs. To transmit environmental stimuli that perceived in the leaves, plants recruit many mobile
molecules including mobile mRNAs as systemic signals for interorgan communication. The mobile
mRNAs provide an efficient and specific remote control system for plants to cope with environmental
dynamics. Upon being transcribed in local tissues, mobile mRNAs are selectively targeted to plasmodesmata for cell-to-cell and long-distance translocation. The mRNA labeling system based on the
RNA-binding protein MS2 provides a useful tool to investigate intracellular trafficking of mobile mRNAs
in plants. Here we describe the detailed protocol to visualize intracellular trafficking of plant mobile
mRNAs by using the MS2 live-cell imaging system.
Key words MS2, Nucleus localization signal, Live-cell imaging, Mobile mRNA, Intracellular
trafficking
1 Introduction
The discovery that mRNA can function non-cell-autonomously has
launched a new era in investigating plant systemic signaling
[1, 2]. These mobile mRNAs are transcribed in local tissues and
delivered cell to cell and long distance to distal organs to regulate
many developmental processes [3–8]. Evidence has suggested that
many mobile mRNAs use phloem as a conduit for long-distance
transport [5, 9, 10]. Extensive analysis of mRNA population in
phloem exudates and interspecies grafting indicated that mobile
mRNAs are widely spread among plant species [11–13]. The pivotal roles of mobile mRNAs in regulating plant development imply
the presence of delicate mechanisms to fine-tune the mobility of
these signaling transcripts [1]. Although computational model
suggests that the movement of mobile mRNAs is determined by
Manfred Heinlein (ed.), RNA Tagging: Methods and Protocols, Methods in Molecular Biology, vol. 2166,
https://doi.org/10.1007/978-1-0716-0712-1_8, © Springer Science+Business Media, LLC, part of Springer Nature 2020
Electronic supplementary material: The online version of this chapter (https://doi.org/10.1007/978-1-07160712-1_8) contains supplementary material, which is available to authorized users.
145
In Vivo Imaging of Mobile mRNAs in Plants Using MS2
Kai-Ren Luo, Nien-Chen Huang, and Tien-Shin Yu
Abstract
Multicellular organisms rely on systemic signals to orchestrate diverse developmental and physiological
programs. To transmit environmental stimuli that perceived in the leaves, plants recruit many mobile
molecules including mobile mRNAs as systemic signals for interorgan communication. The mobile
mRNAs provide an efficient and specific remote control system for plants to cope with environmental
dynamics. Upon being transcribed in local tissues, mobile mRNAs are selectively targeted to plasmodesmata for cell-to-cell and long-distance translocation. The mRNA labeling system based on the
RNA-binding protein MS2 provides a useful tool to investigate intracellular trafficking of mobile mRNAs
in plants. Here we describe the detailed protocol to visualize intracellular trafficking of plant mobile
mRNAs by using the MS2 live-cell imaging system.
Key words MS2, Nucleus localization signal, Live-cell imaging, Mobile mRNA, Intracellular
trafficking
1 Introduction
The discovery that mRNA can function non-cell-autonomously has
launched a new era in investigating plant systemic signaling
[1, 2]. These mobile mRNAs are transcribed in local tissues and
delivered cell to cell and long distance to distal organs to regulate
many developmental processes [3–8]. Evidence has suggested that
many mobile mRNAs use phloem as a conduit for long-distance
transport [5, 9, 10]. Extensive analysis of mRNA population in
phloem exudates and interspecies grafting indicated that mobile
mRNAs are widely spread among plant species [11–13]. The pivotal roles of mobile mRNAs in regulating plant development imply
the presence of delicate mechanisms to fine-tune the mobility of
these signaling transcripts [1]. Although computational model
suggests that the movement of mobile mRNAs is determined by
Manfred Heinlein (ed.), RNA Tagging: Methods and Protocols, Methods in Molecular Biology, vol. 2166,
https://doi.org/10.1007/978-1-0716-0712-1_8, © Springer Science+Business Media, LLC, part of Springer Nature 2020
Electronic supplementary material: The online version of this chapter (https://doi.org/10.1007/978-1-07160712-1_8) contains supplementary material, which is available to authorized users.
145