Utilizing Potato Virus X to Monitor RNA Movement

Yu, Zhiming; Cho, Sung Ki; Zhang, Pengcheng; Hong, Yiguo; Hannapel, David J.

doi:10.1007/978-1-0716-0712-1_10

Cited by 3 publications

References 27 publications

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Spinach-based RNA mimicking GFP in plant cells

Wang

Mei

et al. 2022

Funct Integr Genomics

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Spinach RNA-mimicking GFP (S-RMG) has been successfully used to monitor cellular RNAs including microRNAs in bacterium, yeast, and human cells. However, S-RMG has not been established in plants. In this study, we found that like bacterial, yeast, and human cellular tRNAs, plant tRNAs such as tRNALys can protect and/or stabilize the Spinach RNA aptamer interaction with the fluorophore DFHBI enabling detectable levels of green fluorescence to be emitted. The tRNALys-Spinach-tRNALys, once delivered into “chloroplast-free” onion epidermal cells can emit strong green fluorescence in the presence of DFHBI. Our results demonstrate for the first time that Spinach-based RNA visualization has the potential for in vivo monitoring of RNAs in plant cells.

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Spinach-based RNA mimicking GFP in plant cells

Wang

Mei

et al. 2022

Funct Integr Genomics

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A historical overview of long-distance signalling in plants

Kondhare

Patil

Banerjee

2021

Journal of Experimental Botany

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Be it a small herb or a large tree, intra- and inter-cellular communication as well as long-distance signalling between distant organs is crucial for every aspect of plant development. Vascular system comprising xylem and phloem acts as a major conduit for transmission of long-distance signals in plants. In addition to our knowledge in vascular development, numerous reports in last two decades revealed that selective populations of RNAs, proteins and phytohormones function as mobile signals. Interestingly, many of these signals are shown to regulate diverse physiological processes like flowering, leaf and root development, nutrient acquisition, crop yield and biotic/abiotic stress responses. In this review, we summarize the significant discoveries made in the last 25 years with special emphasis on key mobile signalling molecules (mRNAs, proteins including RNA-binding proteins and small RNAs) that have revolutionized our understanding of how plants integrate various intrinsic and external cues in orchestrating growth and development. Additionally, we provide detailed insights on the emerging molecular mechanisms that possibly control the selective trafficking and delivery of phloem-mobile RNAs to target tissues. We also highlight the cross-kingdom movement of mobile signals during plant-parasite relationships. Considering the dynamic functions of these signals, their implications in crop improvement are also discussed.

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Reappraisal of Spinach-based RNA Visualization in Plants

Mei

Yan

et al. 2020

Preprint

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RNAs can be imaged in living cells using molecular beacons, RNA-binding labeled proteins and RNA aptamer-based approaches. However, Spinach RNA-mimicking GFP (RMG) has not been successfully used to monitor cellular RNAs in plants. In this study, we re-evaluated Spinach-based RNA visualization in different plants via transient, transgenic, and virus-based expression strategies. We found that like bacterial, yeast and human cellular tRNAs, plant tRNAs such as tRNALys (K) can protect and/or stabilize the spinach RNA aptamer interaction with the fluorophore DFHBI enabling detectable levels of green fluorescence to be emitted. The tRNALys-spinach-tRNALys (KSK), once delivered into “chloroplast-free” onion epidermal cells can emit strong green fluorescence in the presence of DFHBI. Transgenic or virus-based expression of monomer KSK, in either stably transformed or virus-infected Nicotinana benthamiana plants, failed to show RMG fluorescence. However, incorporating tandem repeats of KSK into recombinant viral RNAs, enabled qualitative and quantitative detection, both in vitro and ex vivo (ex planta), of KSK-specific green fluorescence, though RMG was less obvious in vivo (in planta). These findings demonstrate Spinach-based RNA visualization has the potential for ex vivo and in vivo monitoring RNAs in plant cells.One sentence summarySpinach-based RMG technology was reevaluated to have potential for ex vivo and in vivo monitoring RNAs in plant cells.

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Utilizing Potato Virus X to Monitor RNA Movement

Cited by 3 publications

References 27 publications

Spinach-based RNA mimicking GFP in plant cells

Spinach-based RNA mimicking GFP in plant cells

A historical overview of long-distance signalling in plants

Reappraisal of Spinach-based RNA Visualization in Plants

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