Diversity in the carotenoid profiles and the expression of genes related to carotenoid accumulation among citrus genotypes

Carotenoids are not only important to the plants themselves but also are beneficial to human health. Since citrus fruit is a good source of carotenoids for the human diet, it is important to study carotenoid profiles and the accumulation mechanism in citrus fruit. Thus, in the present paper, we describe the diversity in the carotenoid profiles of fruit among citrus genotypes. In regard to carotenoids, such as β-cryptoxanthin, violaxanthin, lycopene, and β-citraurin, the relationship between the carotenoid profile and the expression of carotenoid-biosynthetic genes is discussed. Finally, recent results of quantitative trait locus (QTL) analyses of carotenoid contents and expression levels of carotenoid-biosynthetic genes in citrus fruit are shown.     

Interactive effects of leaf age and self-shading on leaf structure,photosynthetic capacity and chlorophyll fluorescence in the rain forest tree,Dryobalanops aromatica

In the tropical canopy tree, Dryobalanops aromatica Gaertn. f., upper-canopy leaves (UL) develop under sunlit conditions but are subjected to self-shading within the crown as they age. In contrast, lower-canopy leaves (LL) are exposed to uniform dim light conditions throughout their life span. By comparing leaf morphology and physiology of UL and LL, variations in leaf characteristics were related to leaf age and self-shading. Mass-based chlorophyll (chl) concentration and the chlorophyll/nitrogen (chl/N) ratio were lower and the chl a/b ratio was higher in UL than in LL. In UL, the chl/N ratio gradually increased and the chl a/b ratio gradually decreased with leaf aging, whereas these ratios remained unchanged with leaf age in LL. The effective quantum yield of photosystem II (PSII) (DeltaF/F(m)') at a given irradiance remained unchanged with leaf age in LL, whereas DeltaF/F(m)' changed with leaf age in UL. These data indicate N reallocation within the leaves from carbon fixation components to light harvesting components and a dynamic regulation of photochemical processes of PSII in response to increased self-shading of UL. Despite the difference in light environment with leaf age between UL and LL, maximum photosynthetic rates and nitrogen-use efficiency decreased with leaf aging in both UL and LL. Constancy in the chl/N ratio with leaf age in LL indicated that the decrease in photosynthetic capacity was caused by effects other than shading, such as leaf aging. We conclude that N reallocation and acclimation of PSII to self-shading occurred even in mature leaves, whereas the change in photosynthetic capacity with leaf age was more conservative.     

Semi-quantitative method to evaluate the α-carbonyl content in lignin

A method to estimate the content of -carbonyl structures in lignin was developed. This method consists of two successive treatments: NaBD₄ treatment of pulp to reduce an -carbonyl structure in lignin, and nitrobenzene oxidation. NaBD₄ was used to convert an -carbonyl structure to a deuterium-labeled hydroxymethine structure. The ratio of D-vanillin [(HO)(H₃CO)C₆H₃CDO] to H-vanillin [(HO)(H₃CO)C₆H₃CHO] or that of their syringyl analogues obtained by nitrobenzene oxidation was used as the measure of the content of -carbonyl structure. Model experiments demonstrated that when sodium hydroxide was used as alkali for the nitrobenzene oxidation, the retention of deuterium at the side chain -position was very low due to the displacement of deuterium with hydrogen by an unknown reaction mechanism. In order to depress this unexpected displacement, the reaction conditions of the nitrobenzene oxidation were modified. The modified nitrobenzene oxidation employs 0.5mol/l of lithium hydroxide as a reaction medium instead of 2.0mol/l sodium hydroxide. By this modification, this method could successfully trace the formation and the degradation of the -carbonyl structure in milled wood lignins.This paper was presented in part at the 11th International Symposium on Wood and Pulping Chemistry, Nice, France, June 2001 and at the 46th Lignin Symposium, Kyoto, Japan, November 2001     

Variations in transpiration rate and leaf cell turgor maintenance in saplings of deciduous broad-leaved tree species common in cool temperate forests in Japan

To clarify mechanisms underlying variation in transpiration rate among deciduous broad-leaved tree species, we measured diurnal changes in stomatal conductance (gs) and leaf water potential, and calculated the maximum transpiration rate (Emax), leaf-specific hydraulic conductance (K(s-l)) and difference between the soil water potential and the daily minimum leaf water potential (Psis - Psi(l,min)). Pressure-volume (P-V) measurements were made on leaves. Saplings of eight broad-leaved tree species that are common in Japanese cool temperate forests were studied. Maximum transpiration rate varied significantly among species. There was a statistically significant difference in Psis - Psi(l,min), but not in K(s-l). Species with large Emax also had large Psis - Psi(l,min) and gs. The results of the P-V analyses showed that species with a large Psis - Psi(l,min) maintained turgor even at low leaf water potentials. The similar daily minimum leaf pressure potentials (Psip) across all eight species indicate that Psip values below this minimum are critical. Based on these results, we suggest that the leaf cell capacity for turgor maintenance strongly affects Psis - Psi(l,min) and consequently Emax via stomatal regulation.     

Lignin fragments rich in detached side-chain structures found in water-soluble LCC

The content of the glyceraldehyde-2-aryl ether-type structure in water-soluble lignin–carbohydrate complex (LCC) fractions, which were obtained from Japanese cedar and birch residual wood meal after milled wood lignin isolation, was determined by ozonation. Quite high amounts of the glyceraldehyde-2-aryl ether-type structure were found in water-soluble LCC fractions of both species; about 3–5 times higher than those of other fractions. This result, as well as the high content of the β-1 structure in these fractions shown in our previous reports, suggest that lignin in these fractions has the characteristics of endwise-type lignin, because the glyceraldehyde-2-aryl ether-type structure and β-1 structure are typical characteristics of this type of lignin fraction. These results are in good agreement with the generally accepted hypothesis that the glyceraldehyde-2-aryl ether-type structure and β-1 structure are generated at the same time by a radical coupling reaction. The results also indicated that these two structures are present in close proximity in the lignin.     

Effects of current-year and previous-year PPFDs on shoot gross morphology and leaf properties in Fagus japonica

We investigated how shoot gross morphology and leaf properties are determined in Fagus japonica Maxim., a deciduous species with flush-type shoot phenology, in which all leaves are produced in a single flush at the start of each season. We examined relationships between current-year shoot properties and local light environment in a 14-m tall beech tree growing in a deciduous forest. Leaf number (LN), total leaf area (TLA), and total leaf length (SL) of the current-year shoot increased with increasing photosynthetic photon flux density (PPFD). Leaf thickness, dry mass per leaf area and nitrogen content on a leaf area basis increased, whereas the chlorophyll/N ratio decreased with increasing PPFD. To separate the effects of current-year PPFD from those of previous year(s), we artificially shaded a part of the uppermost leaf tier. Reciprocal transfers of beech seedlings between controlled PPFD regimes were also made. Characteristics of shoot gross morphology such as LN, TLA and SL were largely determined by the PPFD of the previous year. The exception was the length of the longest "long shoots" with many leaves, in which elongation appeared to be influenced by both previous-year and current-year PPFD. In contrast, leaf properties were determined by current-year PPFD. The ecological implications of our findings are discussed.     

Furazolidone induces the activity of microsomal enzymes that metabolize furazolidone in chickens

The nitrofuran antibacterial agent furazolidone (FZ) is still used in veterinary medicine in some countries in the Middle and Far East. The present study aimed to show the effect of FZ on the activity of microsomal enzymes that metabolize FZ, and to identify the enzyme that contributes to FZ metabolism in chickens. Wistar rats and White Leghorn chickens were administered FZ once a day for four consecutive days. FZ metabolism was accelerated by FZ administration in chickens, but not in rats. The elevation of FZ metabolism coincided with the induction of NADPH cytochrome P450 reductase (CPR) activity in chickens, but such induction was not observed in rats. FZ metabolizing activities were inhibited in the presence of a CPR inhibitor (diphenylene iodonium chloride) but not by the addition of archetypal cytochrome P450 inhibitors (CO or n-octylamine). The preset study concluded that FZ accelerated its own metabolism in the chicken by induction of the activity of CPR.     

Revisiting the mechanism of <Emphasis Type="Italic">β-O</Emphasis>-4 bond cleavage during acidolysis of lignin IV: dependence of acidolysis reaction on the type of acid

The dependence of the acidolysis reaction of a C₆-C₃ dimeric nonphenolic β-O-4 type lignin model compound, 2-(2-methoxyphenoxy)-1-(3,4-dimethoxyphenyl) propane-1,3-diol (veratrylglycerol-β-guaiacyl ether, VG), on the type of acid applied was examined using three different acids [0.2 mol/l HCl, 0.2 mol/l HBr, and 0.1 mol/l (0.2 N) H₂SO₄ in 82% aqueous 1,4-dioxane at 85°C]. In the HCl system, the major reaction modes of the corresponding benzyl cation-type intermediate (BC), which is produced by protonation of the α-hydroxyl group of VG and successive release of the water molecule, are the abstraction of the β-proton and hydride transfer from the β-to the α-position. The liberation of formaldehyde from the γ-hydroxymethyl group of BC is the predominant reaction mode in the H₂SO₄ system. Apparently, an unknown reaction mode or modes is operative in the early stage of the HBr system that causes rapid disappearance of VG accompanied by the quantitative formation of 2-methoxyphenol without affording the common counterpart of a Hibbert’s ketone, 1-hydroxy-3-(3,4-dimethoxyphenyl) propan-2-one. The reaction mode in the HBr system changes with the progress of the reaction and is the same as that in the HCl system after the early stage.     

Effects of Ca2+ and sulfhydryl reductant on the polymerization of soybean glycinin catalyzed by mammalian and microbial transglutaminases

Two types of transglutaminases (TGases), Ca(2+)-dependent TGase derived from guinea pig liver (GTGase) and Ca(2+)-independent TGase derived from a variant of Streptoverticillium mobaraense (MTGase), were used to study the cross-linking of soybean 11S globulin (glycinin). The effects of sulfhydryl reductant (dithiothreitol, DTT) and Ca(2+) on the conformation and TGase-catalyzed polymerization of glycinin were investigated. The conformational change of glycinin was probed by spectral methods. The degree of cross-linking and the polymer (aggregate) formation were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and dynamic light scattering, respectively. Addition of DTT stimulated the TGase-catalyzed cross-linking reactions without destroying the secondary and tertiary structure of glycinin but did not influence the polymer or aggregate formation. It was found that Ca(2+) caused the formation of larger size polymers at lower concentrations, while it suppressed the polymerization at higher concentrations. In addition, the cross-linking behaviors of glycinin were shown to be different between MTGase- and GTGase-catalyzed systems.