Enhancement of saccharification by overexpression of poplar cellulase in sengon

Lignocellulosic material from trees has great potential to form the basis of the second generation for bioethanol production because trees produce most of the biomass on the earth. We modified the wall structure of sengon (Paraserianthes falcataria) through overexpression of poplar cellulase in the cell walls. The overexpression did not decrease cellulose content but caused a decrease in xyloglucan bound to the walls. The level of saccharification and successive ethanol production was increased in the wood of the transgenic sengon overexpressing poplar cellulase compared with that of the wild type plant, and even after delignification of the wood. We propose that a xyloglucan intercalated into cellulose microfibrils could be one of the recalcitrant components in the saccharification of lignocelluloses.     

Proteomic analysis of the G-layer in poplar tension wood

Angiosperm trees bend their stems by forming tension wood at the upper side of leaning stems. Most tension wood has a cellulose-rich G-layer in the innermost surface of the fiber cell wall. Strong tensile stress is considered to occur in the G-layer. This study undertook to identify the proteins involved in G-layer formation and function through a proteomic analysis of G-layer-localized protein. G-layers of poplar were loosened by sonication and isolated as doughnut-shaped pieces of thinly sliced transverse sections. The proteins, once extracted with urea/detergent solution, were separated by two-dimensional polyacrylamide gel electrophoresis, and 110 spots were subjected to liquid chromatography tandem mass spectrometry (LC/MS/MS). A database search for these spots’ mass spectrum patterns identified 72 proteins. In addition, all peptide digestion mixtures of G-layer proteins were separated by strong cation exchange chromatography and 39 proteins were identified using LC/MS/MS analysis. Proteins involved in wall formation, such as lignin biosynthesis-related protein, xyloglucan endotransglucosylase, and fasciclin-like arabinogalactan protein, were notably detected in the G-layer.     

Piezoelectric behavior of wood under combined compression and vibration stresses II: Effect of the deformation of cross-sectional wall of tracheids on changes in piezoelectric voltage in linear-elastic region

This study investigated and clarified the relation between the piezoelectric voltage and microscopic fracture of hinoki (Chamaecyparis obtura Endl.), in particular the deformation of the cross-sectional wall of the tracheid in linear-elastic regions under combined compression and vibration stresses. The piezoelectric voltage-deformation (P-D) curve consisted of a linear region starting from the origin followed by a convex curved region. The linear region of theP-D curve was only about 60% of that of the load-displacement (L-D) curve. By applying combined stresses to a specimen, the cross-sectional walls of the tracheid were deformed mainly at the radial walls. When a tracheid was regarded approximately as a hexagonal prism, the elastic buckling stress of the radial wall was estimated from scanning electron microscope images and our method based on a modification of the Gibson and Ashby method. As a result, it was estimated that the elastic buckling stress was only about 80% of the stress at the proportional limit of theP-D curve. It is found that there are two consecutive regions before the proportional limit of theP-D curve: One is the region up to the spot where the radial cell wall generates the elastic buckling, and the other is the region starting from the end of the aforementioned region up to the proportional limit of theP-D curve.Part of this paper was presented at the 47th annual meeting of the Japan Wood Research Society, Kochi, April 3–5, 1997     

Aggressive behaviour and cannibalism in greater amberjack,Seriola dumerili : effects of stocking density,feeding conditions and size differences

Aggressive behaviour, assessed by 24‐h observations, and survival rates, determined in a 2‐week rearing experiment, were examined in greater amberjack, Seriola dumerili, subjected to single and multiple factors (high density, restricted feeding and diverse size). Aggressive behaviour frequency in the group subjected to both restricted feeding and size diversity (FS) was significantly higher than that in the other groups (Tukey, n=4, P<0.05). In the FS group, aggressive behaviour increased with the starvation time from 19 days post hatching (dph) to 23 dph (two‐way analysis of variance, Tukey, n=4, P<0.05), but decreased gradually thereafter to 46 dph. In the rearing experiment, the survival rates for the diverse size and FS groups were lower than those of the control or restricted feeding groups (Kaplan–Meier, log‐rank test, n=100, P<0.01). Daily mortality in all groups was the highest at 24 dph; although dead fish were usually small, disappearance due to cannibalism was not observed during the rearing period. The results indicate that aggressive behaviour in greater amberjack is induced by hunger and size diversity in the early juvenile stages (23 dph). Mass mortality during seedling production was mainly caused by injury and the death of small fish due to aggressive behaviour/cannibalism.     

Total selenium content of agricultural soils in Japan

To evaluate the selenium (Se) level in agricultural soils in Japan and to investigate its determining factors, 180 soil samples were collected from the surface layer of paddy or upland fields in Japan and their total Se contents were determined. Finely ground soil (50 mg) was wet-digested with HNO₃ and HClO₄ solution and the released Se was reduced to Se(IV). The concentration of Se(IV) was then determined by high-performance liquid chromatography with a fluorescence detector after treatment with 2,3-diaminonaphthalene and extraction with cyclohexane. The total Se content ranged from 0.05 to 2.80 mg kg⁻¹ with geometric and arithmetic means of 0.43 and 0.51 mg kg⁻¹, respectively. The overall data showed a log-normal distribution. In terms of soil type, volcanic soils and peat soils had relatively high Se content and regosols and gray lowland soils had relatively low Se content. In terms of land use, upland soils had significantly higher Se content than paddy soils. Among regions, soils in the Kanto, Tohoku, Hokkaido and Kyushu regions had relatively high content. The total Se content had a significant positive correlation with the organic carbon content ( P  < 0.01) and the equation for the estimation of total Se content with organic carbon suggested that on average approximately 48% (0.24 mg kg⁻¹) of the total Se was in inorganic forms and approximately 52% (0.25 mg kg⁻¹) was in organic forms. Soil pH, on the contrary, did not show a significant relationship with the total Se content. In conclusion, the organic matter content, in combination with volcanic materials, was the main determining factor of the total Se content of agricultural soils in Japan.     

Growth and root sucker ability of field-grown transgenic poplars overexpressing xyloglucanase

Xyloglucan is thought to be a key hemicellulose cross-linking adjacent cellulose microfibrils in plant cell walls. The growth traits of transgenic poplars (Populus alba) with decreased xyloglucan from overexpression of Aspergillus aculeatus xyloglucanase were characterized during a 4-year field trial. The field-trial site consisted of two blocks, a fertile soil block and a non-fertile soil block, determined by soil analysis. In the fertile block, the growth of aboveground biomass of the transgenic poplars was reduced to 24?C44?% compared to that of wild-type poplars, in contrast to the growth seen in chamber and greenhouse conditions. In the non-fertile block, the aboveground biomass of transgenic poplars was also smaller than that of the wild-type poplars. Because poplars reproduce asexually by root suckers, we also compared the formation of root suckers from transgenic and wild-type poplars. Root suckers formed less frequently from transgenic poplars than from wild-type poplars. The growth rates of root suckers from transgenic poplars were also slower than those from wild-type poplars. The results showed that constitutive degradation of xyloglucan impairs poplar growth and vegetative reproduction ability.     

Effects of rice feeding and carnitine addition on growth performance and mRNA expression of protein metabolism‐related genes in broiler grower chicks

This study was carried out to evaluate the nutritional effects of rice feeding and carnitine addition to a diet for broiler chicks. Thirty‐six male 10‐day‐old broiler chicks were assigned to one of the following four treatment groups: corn‐based diet (corn group), rice‐based diet (rice group), and each diet with added carnitine (100 ppm). The experimental period was 2 weeks. Rice feeding resulted in significantly higher growth performance (body weight gain and feed efficiency) compared to corn feeding. Carnitine addition also resulted in higher growth performance. Breast muscle and thigh muscle weight (g) were significantly higher in broiler chicks fed rice and those fed diets with added carnitine. Liver mRNA expression of IGF‐I was significantly higher in broiler chicks fed rice compared to those fed corn. There was no significant difference in mRNA expression of muscle atrogin‐1 or liver CPT‐I between broiler chicks fed rice and those fed corn, not between broilers chicks fed diets containing carnitine or not. Overall, these results show that rice feeding and carnitine addition improve the growth performance of broiler chicks by increasing mRNA expression of liver IGF‐I. In addition, carnitine action is not affected by different cereals (corn and rice).     

Laser induced injury caused hyperglycemia-like effect in Drosophila larva: a possible insect model for posttraumatic diabetes

Diabetic patients need particular care in case of infection, digestive disorder or external injury, because external stress often exasperates the glucose metabolism, which is known as “sick day management”. In addition, severe trauma can be a cause of hyperglycemia with insulin resistance. In spite of critical component of the treatment, the precise mechanisms of how trauma develops posttraumatic diabetes remain unknown. Here, we ablated body wall muscles in Drosophila larvae by laser beam and found that the level of trehalose, the principal sugar circulating in the hemolymph or in the tissues of most insects, was increased. The model may provide a helpful tool to understand the relationship between trauma and sugar metabolism.     

A case of stranded Indo-Pacific bottlenose dolphin (Tursiops aduncus) with lobomycosis-like skin lesions in Kinko-wan,Kagoshima, Japan

Lobomycosis is a chronic fungal disease caused by the etiologic agent, Lacazia loboi, in the skin and subcutaneous tissues in humans and dolphins in tropical and transitional tropical climates. An Indo-Pacific bottlenose dolphin (Tursiops aduncus) stranded in Kagoshima, Japan, had severe skin lesions characterized by granulomatous reactions and hyperkeratosis that were similar to those of the lobomycosis, but no fungal organism was observed in the skin lesion. In this paper, we report a stranded Indo-Pacific bottlenose dolphin with lobomycosis-like lesions based on pathological examinations in Japan.     

In vitro growth of bovine oocytes in oocyte-cumulus cell complexes and the effect of follicle stimulating hormone on the growth of oocytes

Several successful in vitro culture experiments have used oocyte-cumulus cell-mural granulosa cell complexes (OCGCs) from early antral follicles (0.5–0.7 mm) for the growth of bovine oocytes. However, in studies related to in vitro oocyte maturation and in vitro embryo production, oocyte-cumulus cell complexes (OCCs) that have no mural granulosa cells have been widely used instead of OCGCs. The purpose of this study was to determine whether cumulus cells alone support oocyte growth. First, OCCs and OCGCs were cultured in vitro for 14 days to compare the integrity of the complexes as well as antrum formation. After 14 days, the diameter and meiotic competence of oocytes in OCCs and OCGCs were examined. Oocytes in OCCs grew fully and acquired meiotic competence similar to OCGCs, whereas antrum formation occurred later in OCCs as compared to OCGCs. Subsequently, the effects of follicle stimulating hormone (FSH) on in vitro growth of OCCs were examined for 14 days. When FSH was added to the culture medium, OCCs formed antrum-like structures one day earlier than those cultured without FSH. Oocytes cultured with 1 mIU/ml FSH grew fully and acquired meiotic competence. In contrast, when oocytes were cultured in media containing high concentrations of FSH, some of the OCCs collapsed and the number of degenerated oocytes increased. In conclusion, bovine oocytes in OCCs grow and acquire meiotic competence similar to OCGCs and, 1 mIU/ml FSH supports the development of OCCs and oocyte growth as observed in our culture system.