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.     

Improvement of fermentable sugar yields of mangium through transgenic overexpression of xyloglucanase

Recalcitrance to saccharifi cation is a major limiting factor of the conversion of lignocellulosic biomass to ethanol. Levels of wood saccharification and subsequent ethanol production were higher in transgenic mangium (Acacia mangium) trees overexpressing xyloglucanase than in wild-type plants, even after delignification of the wood. We propose that a decrease in the quantity of xyloglucan that is intercalated into cellulose microfibrils could facilitate the process of saccharification.     

Effect of fluvial and geomorphic disturbances on habitat segregation of tree species in a sedimentation-dominated riparian forest in warm-temperate mountainous region in southern Japan

We investigated habitat segregation and patterns of species diversity of trees in relation to variations in fluvial and geomorphic disturbances (erosion or sedimentation) along a longitudinal stream gradient from V-shaped valley to alluvial fan and between valley and adjacent hill slopes in a warm-temperate mountainous riparian forest in Kyushu, southwestern Japan. We longitudinally divided the riparian area into four geomorphic zones: V-shaped valley (VV-zone), upper fan (UF-zone), middle fan (MF-zone), and lower fan (LF-zone). We surveyed the distribution of tree species (diameter at breast height ≥3 cm) in the four riparian zones and in additional plots on hill slopes (SL-zone) representing the broader, nonriparian forest matrix. Detrended correspondence analysis ordination demonstrated variations in species composition along the longitudinal stream gradient. Species guild analysis based on the detection of the species preferred zone by a bootstrap method revealed a guild structure corresponding to each geomorphic zone. The four riparian zones were differentiated from the SL-zone by having a low proportion of SL-guild species and a high proportion of infrequent species that were characterized by deciduous leaf habit. The LF-zone was the most differentiated and was characterized by low tree density and specialist species established on the flat and unstable soil surface created by frequent deposition of sediment. The UF- and MF-zones were characterized by a high tree density and species richness (particularly of infrequent species) established on the stable ground surface of a high alluvial terrace. Microsite heterogeneity produced by channel formation may also maintain a high species diversity in the riparian zones.     

Babesia microti-like parasites detected in Eurasian red squirrels (Sciurus vulgaris orientis) in Hokkaido, Japan

Six Eurasian red squirrels (Sciurus vulgaris orientis), victims of road traffic found during 2002 and 2004 near the Noppro Forest Park in Ebetsu, Hokkaido, Japan, were examined for the presence of Babesia parasites. Three of the six squirrels exhibited positive signals by nested PCRs targeting both the 18S rRNA and beta-tubulin genes. Three squirrels proved to be infected with a B. microti-like parasite as evidenced by sequencing the amplified DNAs and by the morphology of the intraerythrocytic parasites. Genotypically, however, the parasite appeared to be of a new type, as it was clearly distinguishable from any of the known types that have previously been reported in various wild animals. This is the first report showing molecular evidence for the presence of B. microti-like parasites in Sciuridae.     

Ultrasonographic uterine changes and vaginal discharges following intrauterine infusion of liquid paraffin in cows

This study was conducted to examine uterine changes and uterocervical discharges following intrauterine infusion with liquid paraffin (LP) during the luteal phase by ultrasonic and vaginoscopic examinations in cows. Multiparous dairy cows (n=10) were infused with 50 ml physiological saline (PS group; n=5) or liquid paraffin (LP group; n=5) on day 10 or 11 after ovulation (day 0: ovulation). Vaginoscopic, rectal and ultrasonogaphic examinations were carried out at 0.25, 1, 2, 3, 4, 6, 8, 12 and 24 h after the LP and PS infusion and then at daily intervals until subsequent ovulation after the infusions. The mean volumes of recovered discharges from the vagina within 6 h after infusion were significantly greater (P<0.05) in the LP group than in the PS group (33.0 +/- 9.9 vs.14.0 +/- 13.9 ml). Yellowish-white discharge was first observed at 3.2 +/- 0.5 and 3.6 +/- 0.6 h after infusion and lasted for 12.2 +/- 2.9 and 2.1 +/- 1.5 days for the LP and PS groups, respectively, showing a significant difference (P<0.05) in duration. Subsequently, transparent discharge appeared again 2-3 days before the subsequent ovulation after the treatments in both groups and disappeared on the day prior to or the day of ovulation. During the immediate examination after the infusion, the cavity of the uterine horn appeared anechoic and dilated in the images of both groups. The anechoic images changed to echoic images at 2.2 +/- 0.8 and 2.6 +/- 0.9 h after the infusion in the LP and PS groups, respectively, and the echoic images lasted for 12.2 +/- 2.9 and 2.1 +/- 1.5 days in the LP and PS groups, respectively. These results suggest that the appearance and disappearance of intrauterine anechoic and echoic images reflect the appearance and disappearance of the characteristics of the recovered LP/PS-like liquid and yellowish-white and transparent discharges from the vagina.     

Fermentation characteristics and microorganism composition of total mixed ration silage with local food by‐products in different seasons

Total mixed ration (TMR) silage in different seasons was prepared with apple refuse, orchardgrass hay, orchardgrass silage, corn silage and a commercial compound feed in Tohoku, Japan, and with tofu cake, brewers' grains, tall fescue hay, Sudangrass hay, timothy hay, alfalfa hay and a commercial compound feed in Shikoku, Japan, respectively, and their fermentation quality and microorganism composition were evaluated. In Tohoku, the TMR silage in spring, autumn and winter displayed poor quality, with pH value above 4.7, and the harmful microorganisms such as escherichia, yeasts and molds were detected even after 60 days of ensiling; but the TMR silage in summer was well preserved and the escherichia were too few to count after 7 days of ensiling. In Shikoku, the TMR silage in all seasons was well preserved, with low pH, high lactic acid contents, and escherichia, yeasts and molds were consistently at or below the detectable numbers after 7 days of ensiling. The results showed that the TMR silage could not produce sufficient lactic acid to improve silage quality under low‐temperature conditions. Therefore, it is necessary to develop preparation techniques of promoting the lactic acid fermentation for TMR silage in cold seasons.     

Decrease in bovine CD14 positive cells in colostrum is associated with the incidence of mastitis after calving

During the postpartum period there is a high incidence of mastitis in dairy cows. The reason for this increased risk of mastitis still remains unclear. Since leukocytes in colostrum have an important role in preventing the onset of mastitis, we investigated the leukocyte populations, which express CD4, CD8, CD14, CD21 or WC1, in colostrum as well as in blood obtained from 14 Holstein cows. Eight cows developed mastitis within a week after calving and the other 6 remained healthy. The percentage of CD14+ cells in colostrum was significantly lower in mastitic cows than in healthy cows. There were no significant differences in other marker positive cells either in the colostrum or in the blood. The CD14+ cells in colostrum play an important role of defense against invading microorganisms in the mammary glands. Our results suggested that the lower percentage of CD14+ cells in colostrum might predict the incidence of mastitis in the following period.     

Distribution and sequence of pyknotic cells in rat fetuses exposed to busulfan

Busulfan, an antineoplastic bifunctional-alkylating agent, is known to induce developmental anomalies. In the present study, we examined the distribution and sequence of pyknotic cells in rat fetal tissues exposed to busulfan. Pregnant rats on gestation day 13 were administered intraperitoneally 30 mg/kg of busulfan, and fetal tissues were examined at 6, 12, 24, 36, 48, 72 and 96 hours after treatment (HAT). Pyknosis of component cells was observed markedly in the brain, moderately in the eyes and spinal cord and mildly in the craniofacial tissue, mandible, limb buds, tail bud, ganglions, alimentary tract, lungs, kidneys, pancreas and liver. In the brain, mitotic inhibition was also detected. Most of the pyknotic cells were considered to be apoptotic cells judging from the results of TUNEL staining and electron microscopic examination. Commonly in the above-mentioned tissues, pyknotic cells began to increase at 24 HAT, peaked at 36 or 48 HAT and disappeared at 96 HAT, which is when the histological picture returned to normal in most tissues except for the brain, spinal cord and eyes. The present study clarified the outline of busulfan-induced apoptosis in rat fetuses.