Kaori Saito

Direct mapping of morphological distribution of syringyl and guaiacyl lignin in the xylem of maple by time-of-flight secondary ion mass spectrometry

Lignin, one of the main structural polymer of plant cell walls, varies in amount and monomeric composition among tissue and cell types, as well as among plant species. However, few analytical methods are available that can conveniently and accurately determine the morphological distribution of lignin units at the cellular level. In this report, we used time-of-flight secondary ion mass spectrometry (TOF-SIMS) to directly map guaiacyl (G) and syringyl (S) lignin units in several successive growth rings of the maple xylem. TOF-SIMS imaging and a semiquantitative approach revealed clear difference in the annual distribution of lignins between the fiber and vessel. While the vessel walls were constantly G-rich with varied S/G ratios through a growth ring, the fibers showed fairly regular annual distribution of lignins in which the earlywood was S-rich with an almost constant S/G ratio and the latewood was G-rich resulting from a decrease of the S unit. The reliability of TOF-SIMS results was demonstrated by its high correlation with the results of thioacidolysis on radial distribution of the S/G ratio in several contiguous tree rings and also in the latewood and earlywood of each ring. These results indicate that TOF-SIMS allows direct visualization of lignin composition in plant tissues.

Discriminating the Indistinguishable Sapwood from Heartwood in Discolored Ancient Wood by Direct Molecular Mapping of Specific Extractives Using Time-of-Flight Secondary Ion Mass Spectrometry

A new method that can chemically discriminate the visually indistinguishable sapwood from heartwood in discolored woods is presented in this paper. Discriminating between sapwood and heartwood, which are normally recognized by color in cross sections of stems of tress, is important in dendrochronological dating, as well as in evaluating qualities of woods such as durability. In tree-ring chronology, the felling date, which affects the construction date of architectures, can be estimated only in woods that have a recognizable sapwood/heartwood boundary. However, the felling date cannot be estimated in discolored woods because it has indistinguishable sapwood. Time-of-flight secondary ion mass spectrometry (TOF-SIMS) analysis of specific chemical substances retained for approximately 1300 years after felling demonstrated the presence of sapwood in a discolored ancient architectural wood of Hinoki cypress (Chamaecyparis obtusa). Direct molecular mapping by TOF-SIMS clearly indicated that the specific substances, hinokinin, hinokiresinol, hinokione, and hinokiol, started to accumulate at the sapwood/heartwood boundary where only hinokinin was localized and retained predominantly in ray parenchyma cells. The result allowed the determination of the felling date of the discolored wood. TOF-SIMS has shown to be useful for investigating the distribution of minute amounts of chemical components in woods.

Influence of syringyl to guaiacyl ratio on the structure of natural and synthetic lignins.

Several kinds of natural woods and isolated lignins with various syringyl to guaiacyl (S/G) ratios were subjected to thioacidolysis followed by Raney nickel desulfuration to elucidate the relationships between the S/G ratio and the interunit linkage types of lignin. Furthermore, enzymatic dehydrogenation polymers (DHP) were produced by the Zutropf (gradual monolignol addition) method from mixtures of various ratios of coniferyl alcohol and sinapyl alcohol. The analysis of DHPs and natural wood lignins exhibited basically a similar tendency. The existence of both syringyl and guaiacyl units is effective for producing higher amounts of beta-O-4 and 4-O-5 structures, but it lowers the total amount of cinnamyl alcohol and aldehyde end groups. The relative frequency of the beta-beta structure increased, whereas that of beta-5 and 5-5 structures decreased with increasing syringyl units.

Distribution of lignin interunit bonds in the differentiating xylem of compression and normal woods of Pinus thunbergii

The lignification process and lignin distribution at different stages of cell wall differentiation in the secondary xylem of compression and normal woods of Pinus thunbergii were investigated by thioacidolysis and subsequent desulfuration. We prepared 50-µm-thick, contiguous tangential sections of pine shoots, cut from the cambial zone through to mature xylem. In compression wood, uncondensed guaiacyl (G) and p-hydroxyphenyl (H) lignins were deposited simultaneously from early to late stages of lignification. The various types of G-G, G-H, and H-H dimers were detected in compression wood, and the ratio of G-H and H-H dimers to total dimers increased as lignification proceeded. In contrast, uncondensed and condensed H units were detected in trace amounts in normal wood. Significant differences in the relative distributions of lignin interunit linkages were not observed between compression and normal woods or between differentiating and mature xylems in either compression or normal woods.

The accumulation pattern of ferruginol in the heartwood-forming Cryptomeria japonica xylem as determined by time-of-flight secondary ion mass spectrometry and quantity analysis

BACKGROUND AND AIMS Heartwood formation is a unique phenomenon of tree species. Although the accumulation of heartwood substances is a well-known feature of the process, the accumulation mechanism remains unclear. The aim of this study was to determine the accumulation process of ferruginol, a predominant heartwood substance of Cryptomeria japonica, in heartwood-forming xylem. METHODS The radial accumulation pattern of ferruginol was examined from sapwood and through the intermediate wood to the heartwood by direct mapping using time-of-flight secondary ion mass spectrometry (TOF-SIMS). The data were compared with quantitative results obtained from a novel method of gas chromatography analysis using laser microdissection sampling and with water distribution obtained from cryo-scanning electron microscopy. KEY RESULTS Ferruginol initially accumulated in the middle of the intermediate wood, in the earlywood near the annual ring boundary. It accumulated throughout the entire earlywood in the inner intermediate wood, and in both the earlywood and the latewood in the heartwood. The process of ferruginol accumulation continued for more than eight annual rings. Ferruginol concentration peaked at the border between the intermediate wood and heartwood, while the concentration was less in the latewood compared with the earlywood in each annual ring. Ferruginol tended to accumulate around the ray parenchyma cells. In addition, at the border between the intermediate wood and heartwood, the accumulation was higher in areas without water than in areas with water. CONCLUSIONS TOF-SIMS clearly revealed ferruginol distribution at the cellular level. Ferruginol accumulation begins in the middle of intermediate wood, initially in the earlywood near the annual ring boundary, then throughout the entire earlywood, and finally across to the whole annual ring in the heartwood. The heterogeneous timing of ferruginol accumulation could be related to the distribution of ray parenchyma cells and/or water in the heartwood-forming xylem.

Fragmentation mechanism of the phenylcoumaran-type lignin model compound by ToF-SIMS

Abstract The experiments with model compounds revealed that time-of-flight secondary ion mass spectrometry (ToF-SIMS) is able to split the common interunit linkages of lignin, except the 5-5 linkage. In a previous study, ToF-SIMS produced characteristic secondary ions with m/z 137 and 151 (C6-C1 fragments) from the phenylcoumaran-type lignin model compound, the benzofuran ring of which has a β-5 linkage and an α-O-4 linkage. However, it is still unclear whether the fragments are from ring A with the free phenolic OH as a result of the sole cleavage of the α-O-4 link by opening the furan ring or from ring B as a result of the β-5 cleavage. In this study, the phenylcoumaran-type lignin model compound and its deuterium- and/or 13C-labeled analogues were synthesized and analyzed by ToF-SIMS. It could be clarified that the β-5 linkage is not cleaved by ToF-SIMS.

Aluminum localization in the cell walls of the mature xylem of maple tree detected by elemental imaging using time-of-flight secondary ion mass spectrometry (TOF-SIMS)

Abstract The distribution of aluminum (Al) and other inorganic elements has been mapped at the cellular level in the mature xylem of maple by means of time-of-flight secondary ion mass spectrometry (TOF-SIMS) imaging. This method permits to image inorganic elements along with organic molecular species with high spatial resolution, and it was demonstrated that Na, K, Ca, and Mg are almost uniformly distributed in several contiguous growth rings of the air-dried wood sample. In contrast, Al was accumulated heterogeneously and intensely in the cell walls of the secondary xylem, including several growth rings, despite the long-term air-drying during storage. These results are interpreted that Al is immobilized within the cell wall, or it is closely interacted with the components of the secondary xylem. TOF-SIMS mapping also shows that Al has a tendency to be localized around vessels in the inner rings.

Distribution of extracts including 4,8-dihydroxy-5-methoxy-2-naphthaldehyde in Diospyros kaki analyzed by gas chromatography-mass spectrometry and time-of-flight secondary ion mass spectrometry

Abstract The distribution of ethyl acetate extracts and 4,8-dihydroxy-5-methoxy-2-naphthaldehyde (compound I), which is a major constituent of the extracts obtained from the blackened heartwood of Diospyros kaki, was analyzed via gas chromatography-mass spectrometry (GC-MS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS). According to GC-MS, the extracts and compound I are high in concentration at the pith and at the edges of the blackened heartwood. ToF-SIMS analysis revealed a peak at a mass-to-charge ratio of (m/z) 218, which is characteristic of the ionic form of compound I. The ToF-SIMS imaging of compound I in the blackened heartwood based on m/z 218 shows that compound I is located in parenchyma cells and their neighboring axial elements.

Distribution Analysis of Triglyceride Having Repair Effect on Damaged Human Hair by TOF-SIMS

This study showed that hydrogenated palm oil was an effective conditioning agent for repair of damaged hair because it inhibits water-induced swelling of damaged hair. To understand the functional mechanism of hydrogenated palm oil, we conducted TOF-SIMS analysis of the penetration of this oil into damaged hair. TOF-SIMS analysis revealed that tripalmitin, a constituent of hydrogenated palm oil, penetrated into the cuticle and the outer cortex of damaged hair. This is considered as the mechanism responsible for inhibition of hair swelling by hydrogenated palm oil.

Effect of Deuterium Exchange in Lignin on Its Structural Analysis Using FT-Raman Spectroscopy

In lignin structural analysis, 1H-nuclear magnetic resonance (NMR) spectroscopy is frequently applied using dehydrogenative polymer (DHP) [1]. However, this technique has a disadvantage of long measurement time. On the contrast, Fourier transform (FT) Raman spectroscopy has an advantage of short measurement time and gives the information on various molecule structures similar to NMR. However, peak assignment on the FT-Raman spectrum is difficult because of the peak shift by the surrounding environment for the target structure.