Insight into plant cell wall chemistry and structure by combination of multiphoton microscopy with Raman imaging
. Journal of Biophotonics 2018
, e201700164. Publisher's VersionAbstract
Spontaneous Raman scattering microspectroscopy, second harmonic generation (SHG) and 2-photon excited fluorescence (2PF) were used in combination to characterize the morphology together with the chemical composition of the cell wall in native plant tissues. As the data obtained with unstained sections of Sorghum bicolor root and leaf tissues illustrate, nonresonant as well as pre-resonant Raman microscopy in combination with hyperspectral analysis reveals details about the distribution and composition of the major cell wall constituents. Multivariate analysis of the Raman data allows separation of different tissue regions, specifically the endodermis, xylem and lumen. The orientation of cellulose microfibrils is obtained from polarization-resolved SHG signals. Furthermore, 2-photon autofluorescence images can be used to image lignification. The combined compositional, morphological and orientational information in the proposed coupling of SHG, Raman imaging and 2PF presents an extension of existing vibrational microspectroscopic imaging and multiphoton microscopic approaches not only for plant tissues.
Structural Principles in the Design of Hygroscopically Moving Plant Cells
. In Plant Biomechanics: From Structure to Function at Multiple Scales
; Plant Biomechanics: From Structure to Function at Multiple Scales; Springer International Publishing: Cham, 2018; pp. 235–246. Publisher's VersionAbstract
Plants do not have mineralized skeletons. Instead, each of the plant's cells has an envelope of a cellulose-based wall, which provides a mechanical support to the organism. This stiff wall enables plants to assume flexible body shapes. However, the wall interferes with proteinous muscle-like movements of cells and organs because it is too stiff to yield to forces generated by motor proteins. Nevertheless, plants move constantly. The movements rely on water translocations, which result in the swelling (or growth) of cells located strategically. Water may swell protoplasts in movements that require live cells, like tip growth, tropism, and gas exchange. Other movements are initiated by the swelling of cell walls. These occur in dead tissues that can afford drying. The hygroscopically based movement is very common in seed dispersal mechanisms. The seed that detaches from the mother plant is carried by a cellulosic device. This device was synthesized by the plant and programmed to do some mechanical work, like jumping, crawling, and sowing, in order to deliver the seed to a germination location. This nonliving device provides the seed with means to move away from its mother and siblings. The movement may utilize several types of cells, which differ in the arrangement of cell wall cellulose microfibrils. I present here three types of contracting cells that, together with stiff fiber cells resisting any contraction, create a variety of hygroscopic movements.
Mapping of cell wall aromatic moieties and their effect on hygroscopic movement in the awns of stork’s bill
, 3827 - 3841. Publisher's VersionAbstract
The awn in stork’s bill (Erodium gruinum) seed dispersal units coils as it dries. This hygroscopic movement promotes the dissemination and sowing of the seeds. Here we aimed to understand the movement rate, by correlating water dynamics within the awn to the spatial variation in the chemical composition of the awn’s cell walls. We followed the hygroscopic movement visually and measured the kinetics of water adsorption–desorption in segments along the awn. We integrated data from white light, fluorescence, and Raman microscopy, and Matrix Assisted Laser Desorption Ionization imaging to characterize the micro chemical makeup of the awn. We hydrolyzed awns and followed the change in the cell walls’ composition and the effect on the movement. We found that the coil’s top segment is more sensitive to humidity changes than the coil’s base. At the top part of the coil, we found high concentration of modified lignin. In comparison, the base part of the awn contained lower concentration of mostly unmodified lignin. Ferulic acid concentration increased along the awn, apparently cross-linking hemicellulose and strengthening cell-to-cell adhesion. We propose that the high concentration of modified lignin at the coil’s top increased the hydrophobicity of the cell walls, allowed faster water molecules dynamics; thus inducing fast reaction to ambient humidity. Strong cell-to-cell adhesion in this region created a durable tissue required for the awn’s repeated movement that is induced by the diurnal humidity cycles.
Interplay between silica deposition and viability during the life span of sorghum silica cells. New Phytol 2018
Silica cells are specialized epidermal cells found on both surfaces of grass leaves, with almost the entire lumen filled with solid silica. The mechanism precipitating silicic acid into silica is not known. Here we investigate this process in sorghum (Sorghum bicolor) leaves. Using fluorescent confocal microscopy, we followed silica cells' ontogeny, aiming to understand the fate of vacuoles and nuclei. Correlating the confocal and scanning electron microscopy, we timed the initiation of silica deposition in relation to cell's viability. Contrary to earlier reports, silica cells did not lose their nucleus before silica deposition. Vacuoles in silica cells did not concentrate silicic acid. Instead, postmaturation silicification initiated at the cell periphery in live cells. Less than 1% silica cells showed characteristics of programmed cell death in the cell maturation zone. In fully elongated mature leaves, 2.4% of silica cells were nonsilicified and 1.6% were partially silicified. Silica deposition occurs in the paramural space of live silica cells. The mineral does not kill the cells. Instead, silica cells are genetically programmed to undergo cell death independent of silicification. Fully silicified cells seem to have nonsilicified voids containing membrane remains after the completion of the cell death processes.