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1.
Sci Rep ; 5: 13425, 2015 Sep 03.
Article in English | MEDLINE | ID: mdl-26333536

ABSTRACT

The limitations of conventional extracellular recording and intracellular recording make high-resolution multisite recording of plant bioelectrical activity in situ challenging. By combining a cooled charge-coupled device camera with a voltage-sensitive dye, we recorded the action potentials in the stem of Helianthus annuus and variation potentials at multiple sites simultaneously with high spatial resolution. The method of signal processing using coherence analysis was used to determine the synchronization of the selected signals. Our results provide direct visualization of the phloem, which is the distribution region of the electrical activities in the stem and leaf of H. annuus, and verify that the phloem is the main action potential transmission route in the stems of higher plants. Finally, the method of optical recording offers a unique opportunity to map the dynamic bioelectrical activity and provides an insight into the mechanisms of long-distance electrical signal transmission in higher plants.


Subject(s)
Action Potentials/physiology , Helianthus/physiology , Plant Stems/physiology , Voltage-Sensitive Dye Imaging/instrumentation , Voltage-Sensitive Dye Imaging/methods , Equipment Design , Equipment Failure Analysis , Reproducibility of Results , Sensitivity and Specificity
2.
Plant Mol Biol ; 69(3): 313-24, 2009 Feb.
Article in English | MEDLINE | ID: mdl-19002591

ABSTRACT

Nine genes that encode proteins of the MAP65 family have been identified in the Arabidopsis thaliana genome. In this study, we reported that AtMAP65-2, a member of the AtMAP65 family, could strongly stabilize microtubules (MTs). Bacterially-expressed AtMAP65-2 fusion proteins induced the formation of large MT bundles in vitro. Although AtMAP65-2 showed little effect on MT assembly or nucleation, AtMAP65-2 greatly stabilized MTs that were subjected to low-temperature treatment in vitro. Analyses of truncated versions of AtMAP65-2 indicated that the region that encompassed amino acids 495-578, which formed a flexible extended loop, played a crucial role in the stabilization of MTs. Analysis of suspension-cultured Arabidopsis cells that expressed the AtMAP65-2-GFP fusion protein showed that AtMAP65-2 co-localized with MTs throughout the cell cycle. Cortical MTs that were decorated with AtMAP65-2-GFP were more resistant to the MT-disrupting drug propyzamide and to ice treatment in vivo. The results of this study demonstrate that AtMAP65-2 strongly stabilizes MTs and is involved in the regulation of MT organization and dynamics.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis Proteins/physiology , Microtubule-Associated Proteins/metabolism , Microtubule-Associated Proteins/physiology , Microtubules/metabolism , Arabidopsis Proteins/genetics , Cold Temperature , Gene Expression Regulation, Plant/genetics , Gene Expression Regulation, Plant/physiology , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Microscopy, Confocal , Microtubule-Associated Proteins/genetics
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