Sachie Yagami

Distribution of coniferin in freeze-fixed stem of Ginkgo biloba L. by cryo-TOF-SIMS/SEM.

To clarify the role of coniferin in planta, semi-quantitative cellular distribution of coniferin in quick-frozen Ginkgo biloba L. (ginkgo) was visualized by cryo time-of-flight secondary ion mass spectrometry and scanning electron microscopy (cryo-TOF-SIMS/SEM) analysis. The amount and rough distribution of coniferin were confirmed through quantitative chromatography measurement using serial tangential sections of the freeze-fixed ginkgo stem. The lignification stage of the sample was estimated using microscopic observations. Coniferin distribution visualized at the transverse and radial surfaces of freeze-fixed ginkgo stem suggested that coniferin is stored in the vacuoles, and showed good agreement with the assimilation timing of coniferin to lignin in differentiating xylem. Consequently, it is suggested that coniferin is stored in the tracheid cells of differentiating xylem and is a lignin precursor.

Lignification of ray parenchyma cells (RPCs) in the xylem of Phellodendron amurense Rupr.: quantitative and structural investigation by TOF-SIMS and thioacidolysis of laser microdissection cuts of RPCs

Abstract Lignification of ray parenchyma cells (RPCs) in the sapwood (sW), transition zone (TZ), and heartwood (hW) of Phellodendron amurense Rupr. has been investigated by time-of-flight secondary ion mass spectrometry (TOF-SIMS) and thioacidolysis. The results of TOF-SIMS indicate that the relative ion intensity of lignin in RPCs increased from sW to hW, while there was almost no difference in the case of axial wood fibers. The ratio of syringyl (S) to guaiacyl (G) lignin units (S/G value) in RPCs was higher than that in wood fibers. Samples containing more RPCs were prepared by laser microdissection (LMD) and analyzed by thioacidolysis, including also the subsequent Raney nickel desulfurization, and the resulting monomers and dimers were quantitatively analyzed by GC/MS. The monomer analysis shows that lignin content and S/G values in RPCs rich samples increased from sW towards hW. The dimer analysis reveals that the S-S dimers are dominant in all types of samples. The results of TOF-SIMS and thioacidolysis of LMD samples are interpreted that the lignification of RPCs progresses from the sW towards the hW and that the chemical structure of lignin in RPCs is different from that in axial elements.

Enzymatic dehydrogenative polymerization of monolignol dimers

The structure and biosynthesis of lignin are not yet fully understood, especially the step following the initial dimerization of monolignol. Liquid chromatograph mass spectrometer (LC–MS) was used to analyze the consumption rates of monolignol dimers formed by β-O-4, β-5, and β-β couplings between coniferyl alcohols in efforts to understand the activity of monolignol dimers in enzymatic dehydrogenative polymerization. We investigated the reaction kinetics in single-component and mixed-component reaction systems containing one and two species of the dimers, respectively. A difference was observed between the consumption rates of the three dimers we tested, and the consumption rate of one dimer in the single-component reaction was different from that in a mixed-component reaction. In qualitative LC–MS analyses, coniferyl alcohol oligomers were detected in the reaction products. Some monolignol tetramers were formed by 5-5 and 5-O-4 coupling between the dimers. The results of this work suggested that monolignol dimers with β-5 and β-β linkages could function as radical mediators in enzyme-catalyzed polymerization.

Lignification of ray parenchyma cells in the xylem of Pinus densiflora. Part II: Microchemical analysis by laser microdissection and thioacidolysis

Abstract The lignification of ray parenchyma cells in the sapwood (sW) and heartwood (hW) of Pinus densiflora was investigated by thioacidolysis and the subsequent Raney nickel desulfuration. The samples rich and less rich in ray parenchyma cells were prepared by laser microdissection (LMD). The whole sections burned randomly by the laser served as the controls. Guaiacyl (G) monomers were detected in all the samples, and p-hydroxyphenyl (H) monomers were detectable only in trace amounts, while syringyl (S) units were absent, as expected in softwood. In sW samples rich in ray parenchyma cells, the yields of G monomers are significantly lower than in the other samples. The various types of G-G and one G-H dimers were detected, and the β-1′, β-5′, and 5-5′ dimers were dominant. The relative distributions of lignin interunit linkages were very similar in all the samples regardless of the abundance of the ray parenchyma cells in the sW or hW tissues.

Analysis of distribution of wood extractives in Gmelina arborea by gas chromatography and time-of-flight secondary ion mass spectrometry

Abstract The aim of the present study was to investigate the extractives of Gmelina arborea stem from the sapwood (sW) to heartwood (hW) including the transition zone (tZ) between them by means of quantitative GC-MS, while the spatial distribution of four typical compounds was analysed by TOF-SIMS. The focus was on gmelinol, paulownin, 7′-O-ethyl arboreol, and β-sitosterol, which were isolated and purified from hW. The four compounds revealed a characteristic distribution pattern corresponding to their key role in biosynthesis and depending on their susceptibility to secondary reactions in the hW.

ECCENTRIC GROWTH AND GROWTH STRESS IN INCLINED STEMS OF GNETUM GNEMON

Gnetum gnemon L. (Gnetales) forms hardwood-like secondary xylem in its trunks and branches although it is a gymnosperm. The present study tested the surface growth stress in relation to anatomical and chemical properties of the secondary xylem in inclined and vertical stems of G. gnemon using morphological and chemical composition analyses. Secondary growth was promoted on the upper half of the cross section in an inclined stem; at the same time, tensile growth stress increased on the upper side and decreased on the lower side of the inclined stem. However, formation of reaction wood fibers was not detected on either side. The microfibril angle was associated with differences in tensile growth stress. Thus, we conclude that negative gravitropism in G. gnemon is caused by a synergistic effect of increased tensile growth stress as well as the promotion of secondary growth on the upper side of the inclined stem. Our results are comparable to the negative gravitropism observed in the family Magnoliaceae, which does not form gelatinous fibers in its tension wood.

Radical transfer system in the enzymatic dehydrogenative polymerization (DHP formation) of coniferyl alcohol (CA) and three dilignols

Abstract No clear picture has yet been elaborated concerning the mechanism of lignin growth, and thus this topic is the focus of the present paper. Namely, the enzymatic dehydrogenative polymerization (DHP formation) of coniferyl alcohol (CA, as a monolignol) and three dilignols and their reaction kinetics were investigated. The dilignols [guaiacylglycerol-β-coniferyl ether (IβO4), dehydrodiconiferyl alcohol (IIβ5), and pinoresinol (IIIββ)] and CA as a monolignol [(3-OCD3)-coniferyl alcohol (CAOCD3)] were synthesized and subjected to enzymatic DHP formation. The dilignol derived from CAOCD3 could be identified by its higher molecular weight in comparison with the starting dilignols (IβO4, IIβ5, and IIIββ). Based on the observed consumption rate of the CA and its dilignols, it was proposed that a radical transfer system exists between the dilignols, which is generated from the CA and the starting substrates.

Sexual and temporal variations in floral scent in the subdioecious shrub Eurya japonica Thunb

Abstract In many flowering plants, floral scents are a significant trait for visitors, playing an important role in attracting pollinators and/or detracting herbivores. The evolution of flowering plants from hermaphroditism to dioecy is often accompanied by sexual dimorphism in floral scent. In this study, floral scents emitted by different sexual morphs of the subdioecious shrub Eurya japonica Thunb. were collected using a dynamic headspace method, and sexual and temporal variations were evaluated by gas chromatography–mass spectrometry (GC–MS). Two volatiles, α‐pinene and linalool, were identified as the major components of floral scents in females, hermaphrodites, and males. The males emit higher amounts of floral scents, particularly α‐pinene, compared to females or hermaphrodites. Floral scents emitted by males generally decrease as flowers enter senescence, whereas those from females or hermaphrodites do not significantly differ. Intraspecific variations in floral scents of subdioecious species provided by this study would contribute to better understanding of sexual dimorphism in floral scent.

Studies on non-phenolic lignans in alkaline cooking

Abstract The stability and chemical reactions of lignans during alkaline cooking is not yet fully clarified although their degradation products may have a considerable effect on the pulp and its bleaching, such as spotting and color contamination. To fill this gap, a phenolic lignan (pinoresinol) and four non-phenolic furofuran lignans (gmelinol, paulownin, eudesmin and sesamin) were submitted to alkaline cooking at various conditions. The effect of cooking duration shows that non-phenolic lignans are more stable during a 3-h reaction time than pinoresinol. After a 24-h reaction, most of the lignans were decomposed or polymerized. The effect of cooking temperature shows that pinoresinol, eudesmin and sesamin are more susceptible to degradation with increasing temperature. The concentration of the alkaline liquors only affect significantly to pinoresinol because of its acidity. It was also observed that pinoresinol, gmelinol and eudesmin give rise to small amounts of vanillin and acetovanillone, while paulownin and sesamin were mainly polymerized or degraded. To confirm the reaction mechanism of the occurrence of acetovanillone in the reaction, 13C-labeled pinoresinol was also submitted to alkaline cooking. It was revealed that the cleavage at β-β′ position occurred during alkaline reaction. The stability of these compounds during alkaline cooking are not affected by the hydroxyl group in the C-β′ position.

The composition and chemical alteration of gums in the vessels of Phellodendron amurense

Abstract An occluding substance (gum) was observed in the vessels of Phellodendron amurense Rupr. and analysed by spectroscopic and chemical methods. Following safranin-alcian blue staining, the gum in sapwood (sW) turned to blue and in heartwood (hW) to red. The gum was studied in situ by UV and Raman microscopies, time-of-flight secondary ion mass spectrometry (TOF-SIMS). The gum was isolated by laser microdissection (LMD) and it was alkali hydrolysed and the degradation products were analysed by GC-MS. The staining experiments, and the UV and Raman microscopies indicated that the major component of the sW gum is constituted of polysaccharides, while in the hW gum the aromatic character is dominating. TOF-SIMS measurements were interpreted as showing the aromatic substances in the hW gum did not contain lignin. The GC-MS analysis revealed the presence of vanillic acid in the degradation products of hW gum.