Delignification of cell walls of <Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis> during alkaline nitrobenzene oxidation

To clarify the behavior of whole lignins in wood cell walls during alkaline nitrobenzene oxidation, the delignification process from cell walls in normal and compression woods of Chamaecyparis obtusa Endl. (Cupressaceae) was observed using ultraviolet and transmission electron microscopies. The lignin content conspicuously decreased to around 10% after 35min in normal wood. The lignin content in compression wood finally leveled off at aroumd 10% after 50min. In gel filtration of oxidation products in ethyl acetate, a high molecular weight fraction was prominent in extracts from the early stage of the reaction. As the oxidation progressed, the high molecular weight fraction became less prominent in both normal and compression wood. Changes in the weights of cell wall residues during reaction indicated that approximately half of the components other than lignin were also removed from the cell walls. This shows that the majority of lignin with relatively high molecular weight is removed from the cell walls together with polysaccharides in the early stage of the reaction and that further oxidative degradation occurs in solution in later stages. Only a small amount of the lignin with low molecular weight could be analyzed by gas chromatography.Parts of this report were presented at the 47th (Kochi, April 1997) and 48th (Shizuoka, April 1998) Annual Meetings of the Japan Wood Research Society, and at the Lignin Symposium, Sapporo, October 1997     

Infrared spectroscopic study of alkaline oxygen treatment of lignin with ATR technique in aqueous state 1: method for determining quantitative spectra of oxygen-degraded lignin

For the fundamental study of oxygen delignification of kraft pulp, structural changes of kraft lignin during alkaline oxygen treatment were investigated with the use of infrared measurement with attenuated total reflectance (ATR) technique. In the neutralized reaction mixture of alkaline oxygen-treated kraft lignin, there is a significant amount of NaCl, so that the spectral changes of water due to the coexistence of NaCl was investigated, and how to remove the huge absorption of NaCl solution is discussed. Sodium vanillate–NaCl solutions were employed as model solutions for the reaction mixture. Partial least square (PLS) regression was applied for the prediction of NaCl concentration, and the spectrum of NaCl solution was subtracted from the spectrum of sodium vanillate–NaCl solution as background measurement. This allowed us to obtain the vanillate spectra free from the absorption of NaCl solution. In addition, the mathematical method for reconstructing the spectrum of NaCl solution is discussed. The spectrum of NaCl solution is reconstructed as the linear combination of basic spectra calculated by singular value decomposition (SVD), and it was subtracted from that of the sodium vanillate–NaCl solution. By this procedure, the vanillate spectra were also obtained quantitatively, as has been demonstrated in PLS regression study. It was also confirmed that the quantitative spectra of high molecular weight fraction of alkaline oxygen-treated kraft lignin were obtained by the use of this reconstruction technique.Parts of this report were presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, Japan, April 2002 and the 12th International Symposium on Wood and Pulping Chemistry, Madison, USA, June 2003     

Contrasting seasonal leaf habits of canopy trees between tropical dry-deciduous and evergreen forests in Thailand

We compared differences in leaf properties, leaf gas exchange and photochemical properties between drought-deciduous and evergreen trees in tropical dry forests, where soil nutrients differed but rainfall was similar. Three canopy trees (Shorea siamensis Miq., Xylia xylocarpa (Roxb.) W. Theob. and Vitex peduncularis Wall. ex Schauer) in a drought-deciduous forest and a canopy tree (Hopea ferrea Lanessan) in an evergreen forest were selected. Soil nutrient availability is lower in the evergreen forest than in the deciduous forest. Compared with the evergreen tree, the deciduous trees had shorter leaf life spans, lower leaf masses per area, higher leaf mass-based nitrogen (N) contents, higher leaf mass-based photosynthetic rates (mass-based P(n)), higher leaf N-based P(n), higher daily maximum stomatal conductance (g(s)) and wider conduits in wood xylem. Mass-based P(n) decreased from the wet to the dry season for all species. Following onset of the dry season, daily maximum g(s) and sensitivity of g(s) to leaf-to-air vapor pressure deficit remained relatively unchanged in the deciduous trees, whereas both properties decreased in the evergreen tree during the dry season. Photochemical capacity and non-photochemical quenching (NPQ) of photosystem II (PSII) also remained relatively unchanged in the deciduous trees even after the onset of the dry season. In contrast, photochemical capacity decreased and NPQ increased in the evergreen tree during the dry season, indicating that the leaves coped with prolonged drought by down-regulating PSII. Thus, the drought-avoidant deciduous species were characterized by high N allocation for leaf carbon assimilation, high water use and photoinhibition avoidance, whereas the drought-tolerant evergreen was characterized by low N allocation for leaf carbon assimilation, conservative water use and photoinhibition tolerance.     

Phyllochron Dynamics Under Controlled Environments in Rice (Oryza Sativa L.)

Previously, we reported that phyllochron of rice greatly fluctuated in the course of development under natural conditions. The present study was carried out to examine the phyllochron dynamics using the five near-isogenic lines (NILs) for heading time and the recurrent parent. The change of phyllochron was investigated under four different controlled conditions regarding daylength and temperature. Growth duration (days to heading) showed a large variation among different growth regimes as well as genotypes. Changes of phyllochron were evaluated in the genotypes by using a quartic polynomial regression that was well fitted to the change of phyllochron in the present controlled environments as previously observed under natural conditions. Phyllochron began to drastically increase after around the 5th phytomer order and decreased with the progress of reproductive development in similar manners under different environments. The pattern of phyllochron change varied from a flat to non-flat shape. The shape of the equation was affected by genotypes as well as daylengths and temperatures. In addition, the shape was closely related to the growth duration (days to heading) of the genotype, suggesting that the phyllochron might internally increase depending on the genotype controlling the heading date. Therefore, the present results showed that the sufficient examination of phyllochron dynamics is necessary for understanding a variation in the heading date of rice.     

Developmental changes of phyllochron in near-isogenic lines of rice (Oryza sativa L.) with different growth durations

The repeated elements called phytomers, which consist of leaf, node, internode and axillary bud, play an important role in the development of modular organization in plants. Rice has the striking feature that the rate of the phytomer production is closely synchronized with the rate of leaf emergence (phyllochron). We examined developmental changes of phyllochron by using 10 near-isogenic lines (NILs) showing diversified growth durations in rice. The NILs were established by backcrosses with a strain practically insensitive to photoperiod, and they consisted of early- or late-flowering NILs whose differences of growth duration were caused by the combinations of alleles at 6 loci. The developmental patterns of phyllochron were evaluated by means of a quartic polynomial, which fitted well in most cases. The results indicated that phyllochron greatly changed during development, especially in late-flowering NILs as well as the recurrent parents, although the fluctuation of phyllochron was not so marked in the early-flowering NILs. Thus, the developmental change of phyllochron was highly dependent upon the genotypes and/or growth duration; however, it was associated with neither floral initiation nor temperature, indicating that the change of phyllochron might reflect internal or physiological changes which occur during the life cycle of rice. This revised version was published online in July 2006 with corrections to the Cover Date.     

Determination of theaflavins including methylated theaflavins in black tea leaves by solid-phase extraction and HPLC analysis

A quantitative method for four theaflavins and two methylated theaflavin derivatives in black tea leaves was developed by solid-phase extraction and a high-performance liquid chromatographic method with photodiode array detection. The theaflavins in black tea leaves were extracted three times with 40 vol 50% aqueous ethanol (mg dry tea powder/mL) containing 2% ascorbic acid. The ethanol extracts were diluted 4-fold with distilled water. All diluted extracts were directly applied to the solid-phase C18 cartridge column without concentration. The fraction of theaflavins was obtained by 40% ethanol extraction after rinsing with water followed with 15% ethanol extraction. An aliquot of theaflavins after concentration was injected onto an ODS C18 reversed-phase column, and four theaflavins and two methylated theaflavins were sufficiently separated by a linear gradient system using distilled water and acetonitrile with 0.5% acetic acid. This analytical method is sensitive for the determination of a small amount of methylated theaflavins, since various interfering substances produced during the fermentation process were eliminated in advance by solid-phase extraction. Using this analytical method, we also demonstrated that methylated theaflavins were easily produced during the manufacture of black tea.     

Antiallergic tea catechin, (-)-epigallocatechin-3-O-(3-O-methyl)-gallate,suppresses FcepsilonRI expression in human basophilic KU812 cells

We previously found that the O-methylated derivative of (-)-epigallocatechin-3-O-gallate (EGCg), (-)-epigallocatechin-3-O-(3-O-methyl)-gallate (EGCG' '3Me), has potent antiallergic activity. The high-affinity IgE receptor, FcepsilonRI, is found at high levels on basophils and mast cells and plays a key role in a series of acute and chronic human allergic reactions. To understand the mechanism of action for the antiallergic EGCG' '3Me, the effect of EGCG' '3Me on the cell surface expression of FcepsilonRI in human basophilic KU812 cells was examined. Flow cytometric analysis showed that EGCG' '3Me was able to decrease the cell surface expression of FcepsilonRI. Moreover, immunoblot analysis revealed that total cellular expression of the FcepsilonRI alpha chain decreased upon treatment with EGCG' '3Me. FcepsilonRI is a tetrameric structure comprising one alpha chain, one beta chain, and two gamma chains. The level of mRNA production of each subunit in KU812 cells was investigated. EGCG' '3Me reduced FcepsilonRI alpha and gamma mRNA levels. The cross-linkage of FcepsilonRI causes the activation of basophils, which leads to the secretion of inflammatory mediators including histamine. EGCG' '3Me treatment inhibited the FcepsilonRI cross-linking-induced histamine release. These results suggested that EGCG' '3Me can negatively regulate basophil activation through the suppression of FcepsilonRI expression.     

Aggregates and gel network structure of globin hydrolysates

A gel with excellent functional properties was prepared successfully using the hydrolysates of globin. In the present study, the structures of intermediate aggregates and gel network were observed directly with an electronic microscope. It was shown clearly that the intermediate aggregates were in a thin rod shape with a length of 130--140 nm, which was in good accordance with the results of the light scattering obtained in a previous study. The diameter of intermediate aggregates was 4--5 nm. Each unit of the intermediate aggregate was composed of beta-chain and peptide beta-1 in a ratio of 1:1. Its molecular weight was 26922 Da, and it had a diameter of 4.1 nm. The thin rod-shaped aggregates were formed with units through the hydrophobic interaction. The length of intermediate aggregate was >30--33 times the diameter. Furthermore, the cross-linked structure formed by peptide alpha-1 and the thin rod-shaped aggregates was also confirmed by the photography of the electronic microscope. These results supported the model proposed in previous papers as proper to depict exactly the formation and structure of the gel network of globin hydrolysates.