Journal ArticleDOI
Transient expression of fluorescent fusion proteins in protoplasts of suspension cultured cells
Yansong Miao,Liwen Jiang +1 more
TLDR
This protocol details an in vivo transient expression system to study the subcellular localization and dynamic associations of plant proteins using protoplasts freshly prepared from Arabidopsis or tobacco BY-2 suspension cultured cells.Abstract:
Transient expression of fluorescent fusion proteins in plant cells has dramatically facilitated our study of newly identified genes and proteins. This protocol details an in vivo transient expression system to study the subcellular localization and dynamic associations of plant proteins using protoplasts freshly prepared from Arabidopsis or tobacco BY-2 suspension cultured cells. The method relies on the transformation of DNA constructs into protoplasts via electroporation. The whole protocol is comprised of three major stages: protoplast generation and purification, transformation of DNA into protoplasts via electroporation and incubation of protoplasts for protein analysis. Similar to stably transformed cell lines, transformed protoplasts are compatible with protein localization studies, pharmaceutical drug treatment and western blot analysis. This protocol can be completed within 11-24 h from protoplast production to protein detection.read more
Citations
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Journal ArticleDOI
The Phosphate Transporter Gene OsPht1;8 Is Involved in Phosphate Homeostasis in Rice
Hongfang Jia,Hongyan Ren,Mian Gu,Jianning Zhao,Shubin Sun,Xiao Zhang,Jieyu Chen,Ping Wu,Guohua Xu +8 more
TL;DR: The data suggest that OsPT8 is involved in Pi homeostasis in rice and is critical for plant growth and development.
Journal ArticleDOI
The novel quantitative trait locus GL3.1 controls rice grain size and yield by regulating Cyclin-T1;3.
Peng Qi,Youshun Lin,Xian-Jun Song,Jinbo Shen,Wei Huang,Jun-Xiang Shan,Mei-Zhen Zhu,Liwen Jiang,Ji-Ping Gao,Hong-Xuan Lin +9 more
TL;DR: The cloning and characterization of a new quantitative trait locus (QTL) for the control of rice grain length, weight and yield is reported, suggesting a new mechanism for the regulation of grain size and yield that is driven through a novel phosphatase-mediated process that affects the phosphorylation of Cyclin-T1;3 during cell cycle progression.
Journal ArticleDOI
Organelle pH in the Arabidopsis Endomembrane System
TL;DR: Two pH sensors generated by combining the improved-solubility feature of solubility-modified green fluorescent protein with the pH-sensing capability of the pHluorins and codon optimized for expression in Arabidopsis are excellent pH sensors for visualization and quantification of pH in vivo, respectively.
Journal ArticleDOI
The Role of a Potassium Transporter OsHAK5 in Potassium Acquisition and Transport from Roots to Shoots in Rice at Low Potassium Supply Levels
Tianyuan Yang,Song Zhang,Yibing Hu,Fachi Wu,Qingdi Hu,Guang Chen,Jing Cai,Ting Wu,Nava Moran,Ling Yu,Guohua Xu +10 more
TL;DR: Results demonstrate that OsHAK5 plays a major role in K acquisition by roots faced with low external K and in K upward transport from roots to shoots in K-deficient rice plants.
Journal ArticleDOI
Multivesicular bodies mature from the trans-Golgi network/early endosome in Arabidopsis.
David Scheuring,Corrado Viotti,Falco Krüger,Fabian Künzl,Silke Sturm,Julia Bubeck,Stefan Hillmer,Lorenzo Frigerio,David G. Robinson,Peter Pimpl,Peter Pimpl,Karin Schumacher +11 more
TL;DR: Evidence is presented that post-TGN transport toward lytic vacuoles occurs independently of clathrin and that MVBs/LEs are derived from the TGN/EE through maturation, indicating that MVB originate from theT GN/EE in a process that requires the action of ESCRT for the formation of intraluminal vesicles and annexins for the final step of releasing MVBs as a transport carrier to the vacuole.
References
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Journal ArticleDOI
Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis.
TL;DR: The transient gene expression system using Arabidopsis mesophyll protoplasts has proven an important and versatile tool for conducting cell-based experiments using molecular, cellular, biochemical, genetic, genomic and proteomic approaches to analyze the functions of diverse signaling pathways and cellular machineries.
Journal ArticleDOI
Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants.
TL;DR: The protocol presented here has underpinned much of the in vivo results highlighting the dynamic nature of the plant secretory pathway and is a relatively fast technique to assess expression of genes of interest.
Journal ArticleDOI
Studying protein dynamics in living cells.
TL;DR: Live cell imaging, in combination with photobleaching, energy transfer or fluorescence correlation spectroscopy are providing unprecedented insights into the movement of proteins and their interactions with cellular components.
Book ChapterDOI
Tobacco BY-2 Cell Line as the “HeLa” Cell in the Cell Biology of Higher Plants
TL;DR: The preparation of protoplasts from TBY-2 cells, from which the isolation of organelles is easy, has been established and using the synchrony system, the change in the cell cycle progression of TBY -2 cells successfully followed thechange in cytoskeletons.
Journal ArticleDOI
Development and use of fluorescent protein markers in living cells.
TL;DR: The development of highly visible and minimally perturbing fluorescent proteins that, together with updated fluorescent imaging techniques, are providing unprecedented insights into the movement of proteins and their interactions with cellular components in living cells are traced.
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