Application of Lifeact Reveals F-Actin Dynamics in Arabidopsis thaliana and the Liverwort, Marchantia polymorpha

Actin plays fundamental roles in a wide array of plant functions, including cell division, cytoplasmic streaming, cell morphogenesis and organelle motility. Imaging the actin cytoskeleton in living cells is a powerful methodology for studying these important phenomena. Several useful probes for live imaging of filamentous actin (F-actin) have been developed, but new versatile probes are still needed. Here, we report the application of a new probe called Lifeact for visualizing F-actin in plant cells. Lifeact is a short peptide comprising 17 amino acids that was derived from yeast Abp140p. We used a Lifeact–Venus fusion protein for staining F-actin in Arabidopsis thaliana and were able to observe dynamic rearrangements of the actin meshwork in root hair cells. We also used Lifeact–Venus to visualize the actin cytoskeleton in the liverwort Marchantia polymorpha; this revealed unique and dynamic F-actin motility in liverwort cells. Our results suggest that Lifeact could be a useful tool for studying the actin cytoskeleton in a wide range of plant lineages

High-Dynamic-Range Spectral Imaging System for Omnidirectional Scene Capture

Omnidirectional imaging technology has been widely used for scene archiving. It has been a crucial technology in many fields including computer vision, image analysis and virtual reality. It should be noted that the dynamic range of luminance values in a natural scene is quite large, and the scenes containing various objects and light sources consist of various spectral power distributions. Therefore, this paper proposes a system for acquiring high dynamic range (HDR) spectral images for capturing omnidirectional scenes. The system is constructed using two programmable high-speed video cameras with specific lenses and a programmable rotating table. Two different types of color filters are mounted on the two-color video cameras for six-band image acquisition. We present several algorithms for HDR image synthesis, lens distortion correction, image registration, and omnidirectional image synthesis. Spectral power distributions of illuminants (color signals) are recovered from the captured six-band images based on the Wiener estimation algorithm. In this paper, we present two types of applications based on our imaging system: time-lapse imaging and gigapixel imaging. The performance of the proposed system is discussed in detail in terms of the system configurations, acquisition time, artifacts, and spectral estimation accuracy. Experimental results in actual scenes demonstrate that the proposed system is feasible and powerful for acquiring HDR spectral scenes through time-lapse or gigapixel omnidirectional imaging approaches. Finally, we apply the captured omnidirectional images to time-lapse spectral Computer Graphics (CG) renderings and spectral-based relighting of an indoor gigapixel image

Higher plant myosin XI moves processively on actin with 35 nm steps at high velocity

High velocity cytoplasmic streaming is found in various plant cells from algae to angiosperms. We characterized mechanical and enzymatic properties of a higher plant myosin purified from tobacco bright yellow-2 cells, responsible for cytoplasmic streaming, having a 175 kDa heavy chain and calmodulin light chains. Sequence analysis shows it to be a class XI myosin and a dimer with six IQ motifs in the light chain-binding domains of each heavy chain. Electron microscopy confirmed these predictions. We measured its ATPase characteristics, in vitro motility and, using optical trap nanometry, forces and movement developed by individual myosin XI molecules. Single myosin XI molecules move processively along actin with 35 nm steps at 7 μm/s, the fastest known processive motion. Processivity was confirmed by actin landing rate assays. Mean maximal force was ∼0.5 pN, smaller than for myosin IIs. Dwell time analysis of beads carrying single myosin XI molecules fitted the ATPase kinetics, with ADP release being rate limiting. These results indicate that myosin XI is highly specialized for generation of fast processive movement with concomitantly low forces