Journal
FRONTIERS IN PLANT SCIENCE
Volume 12, Issue -, Pages -Publisher
FRONTIERS MEDIA SA
DOI: 10.3389/fpls.2021.658836
Keywords
protoplast transient expression; organelle markers; membrane compartments; subcellular localization; protein– protein interaction; live imaging
Categories
Funding
- National Natural Science Foundation of China [31971735]
- Fundamental Research Funds for the Provincial Universities of Zhejiang [2020KJ001]
- Zhejiang Provincial Natural Science Foundation of China [LR20C020001, LZ20C160002]
- National Key Research and Development Program of China [2018YFD1000604]
- Research Grants Council of Hong Kong [R4005-18]
- Zhejiang AF University [2018FR029]
- 111 Project [D18008]
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A protoplast transient gene expression system was established in moso bamboo, generating multicolored fluorescent markers for analyzing subcellular localization and functional study of unknown proteins. This system will promote rapid progress in diverse areas of research in bamboo plants.
Bamboo is an important model plant to study the molecular mechanisms of rapid shoot growth and flowering once in a lifetime. However, bamboo research about protein functional characterization is largely lagged behind, mainly due to the lack of gene transformation platforms. In this study, a protoplast transient gene expression system in moso bamboo has been first established. Using this reliable and efficient system, we have generated a set of multicolored fluorescent markers based on the targeting sequences from endogenous proteins, which have been validated by their comparative localization with Arabidopsis organelle markers, in a combination with pharmaceutical treatments. Moreover, we further demonstrated the power of this multicolor marker set for rapid, combinatorial analysis of the subcellular localization of uncharacterized proteins, which may play potential functions in moso bamboo flowering and fast growth of shoots. Finally, this protoplast transient gene expression system has been elucidated for functional analysis in protein-protein interaction by fluorescence resonance energy transfer (FRET) and co-immunoprecipitation analysis. Taken together, in combination with the set of moso bamboo organelle markers, the protoplast transient gene expression system could be used for subcellular localization and functional study of unknown proteins in bamboo and will definitely promote rapid progress in diverse areas of research in bamboo plants.
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