Genetic variation in resistance to blast disease (Pyricularia oryzae Cavara) in Japanese rice (Oryza sativa L.), as determined using a differential system

A total of 324 Japanese rice accessions, including landrace, improved, and weedy types were used to 1) investigate genetic variations in blast resistance to standard differential isolates, and 2) across the genome using polymorphism data on 64 SSR markers. From the polymorphism data, the accessions were classified into two clusters. Accessions from irrigated lowland areas were included mainly in cluster I, and upland and Indica types were mainly in cluster II. The accessions were classified into three resistance subgroups, A2, B1 and B2, based on the reaction patterns to blast isolates. The accessions in A2 were postulated to have at least two resistance genes Pish and Pik-s, whereas those in B1 had various combinations of the resistance genes Pish, Pia, Pii, Pi3, Pi5(t), and Pik alleles. The B2 accessions were resistant to almost all isolates, and many accessions of cluster II were included, and had Pish, Pia, Pii, Pi3, Pi5(t), certain Pik, Piz and Pita alleles, and unknown genes. The frequencies of accessions of B1 originating in Hokkaido, and those of B2 originating in the Kanto and Tohoku regions were remarkably higher than in the other regions.     

Genotypic variations in the plasticity of nodal root penetration through the hardpan during soil moisture fluctuations among four rice varieties

Rainfed lowland rice fields are characterized by soil moisture fluctuations (SMF) and the presence of hardpan that impedes deep rooting and thus limits water extraction from deep soil layer during the periods of drought. In this study, we used rootboxes with three layers; shallow layer, artificial hardpan, and deep and wet layer below the hardpan, to evaluate differences in the plasticity of nodal roots elongation through the hardpan and promote root branching below the hardpan in response to SMF among four rice varieties; Sasanishiki, Habataki, Nipponbare, and Kasalath. Experiments were conducted during the summer and autumn seasons. Plasticity was computed as the difference in root traits within each variety between the SMF and continuously well-watered treatments. In both experiments, Habataki consistently tended to exhibit higher root plasticity than the other three varieties by increasing number of nodal roots that penetrated the hardpan during rewatering period in SMF, when the soil moisture increased and penetration resistance decreased. This root plasticity then contributed to greater water use at the deeper soil during the subsequent drought period and overall shoot dry matter production. Habataki had significantly higher δ¹³C value in roots at deep layer than roots at the shallow and hardpan layers under SMF, which may indicate that these were relatively newly grown roots as a consequence of root plasticity. This study also indicates that CSSLs derived from Sasanishiki and Habataki varieties may be suitable for the analysis of QTLs associated with root plasticity expression in rainfed lowland with hardpan and experiencing SMF.     

Characterization of lignin-derived products from various lignocellulosics as treated by semi-flow hot-compressed water

To elucidate the decomposition behaviors of lignin from different taxonomic groups, five different lignocellulosics were treated with hot-compressed water (230 °C/10 MPa/15 min) to fractionate lignins into water-soluble portions, precipitates, and insoluble residues. The lignin-derived products in each fraction were characterized and compared. The delignification of monocotyledons [nipa palm (Nypa fruticans) frond, rice (Oryza sativa) straw, and corn (Zea mays) cob] was more extensive than that achieved for Japanese cedar (Cryptomeria japonica, gymnosperm) and Japanese beech (Fagus crenata, dicotyledon angiosperm). The water-soluble portions contained lignin monomers like coniferyl alcohol and phenolic acids, while the precipitates contained higher molecular weight lignin with high content of ether-type linkages. Lignin in the insoluble residues was rich in condensed-type structures. In all five lignocellulosics, ether-type linkages were preferentially cleaved, while condensed-type lignin showed resistance to hot-compressed water. In the monocotyledons, lignin–carbohydrate complexes were cleaved and gave lignins that had higher molecular weights than those eluted from the woods. These differences would facilitate the delignification in monocotyledons. Such information provides useful information for efficient utilization of various lignocellulosics.     

Complete gasification of cellulose in glow-discharge plasma

The behaviors of cellulose (commercially available filter paper) were investigated in glow-discharge plasma, where pyrolysis does not occur because of low temperatures. Cellulose filter papers were decomposed in a glow-discharge plasma with nitrogen flow even at low temperatures of around 50 °C and disappeared completely after a treatment of approximately 90 h without char formation. Tar formation was not observed on any surfaces inside the plasma chamber and vacuum lines. Hence, it was concluded that all cellulose was decomposed into gaseous products. An in-situ analysis of gaseous products by quadrupole mass spectrometry suggested the formation of H₂, H₂O, CO and CO₂ from cellulose. These findings indicate that a clean and complete gasification of cellulose can be achieved with glow-discharge plasma.     

Effect of nutrient levels during the far‐off period on postpartum productivity in dairy cows

The present study aimed to determine the effects of differing nutrient levels during the far‐off period on postpartum metabolism and milk production in lactating cows. Twenty‐six multiparous cows were assigned to three dietary treatments in the far‐off period: a low‐energy diet (L, n = 9, 80% intake of the total digestible nutrients requirement), a moderate‐energy diet (M, n = 8, 105%) and a high‐energy diet (H, n = 9, 130%). During the close‐up period, all cows were provided with 105% intake. After parturition, all cows were fed a lactation diet. The BCS recovery was slow, and low milk yield was found in the H group. In the L group, BCS recovery was favorable after parturition, and lactation persistence was increased. The L group had low rumen endotoxin activity and a high initial ovulation rate after parturition. These findings indicate that a high‐energy diet during the far‐off period has a deleterious effect on milk production. In contrast, the restricted diet in the far‐off period increased adaptability with respect to peri‐parturition metabolic changes, improved the post‐parturition nutritional state, and increased milk production. Furthermore, it suggests that the nutrient levels in the far‐off period affect rumen endotoxin activity and reproductive function after parturition.     

Rhodococcus equi virulence plasmids recovered from horses and their environment in Jeju, Korea: 90-kb type II and a new variant, 90-kb type V

Rhodococcus equi was isolated from fecal and soil samples from four native Jeju horse farms and six Thoroughbred farms in Jeju, Korea. The isolates were examined for the presence of virulence-associated 15-17-kDa antigens (VapA) by colony blotting, using the monoclonal antibody 10G5, and for the gene encoding VapA by PCR. R. equi was isolated from all 36 soil samples collected from the 10 farms with between 5.0 x 10(2) and 7.5 x 10(4) colony-forming units (cfu) per gram of soil, and from 37 of 40 fecal samples with between 5.0 x 10(1) and 1.1 x 10 (5) cfu per gram of feces. Virulent R. equi was isolated from seven farms and appeared in 2.0% of isolates (10 of 508). Of the 10 virulent isolates, four contained a 90-kb type II plasmid, which has been found in isolates from the Kiso native horses of Japan, and the other six contained a new variant, which did not display the EcoRI and EcoT22I digestion patterns of the 10 representative plasmids already reported (85-kb types I, II, III, and IV; 87-kb types I and II; 90-kb types I, II, III, and IV). We designated the new variant as the "90-kb type V" plasmid, because its EcoRI digestion pattern is similar to that of the 90-kb type II plasmid. This is the first report of the prevalence of virulent R. equi in Jeju, Korea. The same virulence plasmid type is found in both Korean and Japanese isolates, providing insight into the origin, ancestry, and dispersal of native horses in Korea and Japan.     

Flow cytometric analysis of peripheral blood mononuclear cells induced by experimental endotoxemia in horse

Cellular activation and functional cell surface markers were evaluated during experimentally-induced endotoxemia in healthy horses. Eight healthy adult horses were infused a low dose of endotoxin (lipopolysaccharide from Escherichia coli O26: B6, 30 ng/kg of body weight, IV) and five control horses were given an equivalent volume of sterile saline solution. Venous blood samples were collected for flow cytometric analysis of peripheral blood mononuclear cells (PBMCs) and to measure plasma endotoxin concentrations. Clinical signs of endotoxemia were recorded at 10, 20, 30, 40, 50 min, 1, 2, 3, 4, 8, 16, 24 and 48 hr after endotoxin or saline solution administration. Clinical findings characteristic of endotoxemia (tachycardia, tachypnea, increased rectal temperature, and leukopenia) occurred transiently in all horses administered endotoxin; however, plasma endotoxin concentrations were detectable in only 50% (4/8) of the endotoxin-infused horses. The percentage of CD4(+), CD5(+), and CD8(+) cells decreased while the percentage of CD14(+), IgM(+), and MHC class II(+) cells increased significantly after endotoxin infusion. Alterations in the immunophenotype of PBMCs from horses with experimentally-induced endotoxemia were associated with changes in vital signs, indicating that endotoxin altered the immuno balance.     

Molecular epidemiology of VapA-positive Rhodococcus equi in thoroughbred horses in Kagoshima,Japan

The prevalence of virulent R. equi having 15- to 17-kDa antigens (VapA) in fecal isolates from 13 thoroughbred foals and their dams on 5 farms in Kagoshima, Japan, and the plasmid profiles of VapA-positive isolates by restriction fragment digestion patterns were investigated to compare the genotypic variation among virulence plasmids of R. equi isolates from Japan. In total, 218 (24.6%) of 886 isolates from the feces of the 13 foals and 13 (12.5%) of 104 isolates from the feces of their dams demonstrated VapA-positive R. equi. Plasmid DNA preparations of 231 virulent isolates from foals and dams were analyzed by restriction enzyme digestion with endonucleases EcoRI, EcoT22I and HindIII and were divided into 3 types: 172 isolates contained a 90-kb type I plasmid, 57 contained a 90-kb type III plasmid and 2 contained a 90-kb type IV plasmid. This study demonstrates a geographic character in the distribution of virulence plasmids found in VapA-positive isolates from thoroughbred foals in Kagoshima.     

Somaclonal variation in Tricyrtis hirta plants regenerated from 1-year-old embryogenic callus cultures

The Liliaceous perennial Tricyrtis hirta, sometimes called ‘Japanese toad lily’, has recently become popular as an ornamental for pot and garden uses. Highly embryogenic callus cultures of this plant predominately consisted of diploid cells but also contained tetraploid cells after 1 year of establishment. In the present study, plans regenerated from the 1-year-old embryogenic callus cultures were subjected to ploidy level analysis and morphological characterization following 3 years of cultivation. Among 37 plants examined, 28 kept the diploid level (2n = 2x = 26) but nine were tetraploid (2n = 4x = 52) as indicated by FCM analysis and chromosome observation. Although no morphological alterations were detected in 26 out of 28 diploid regenerants, the remaining two showed noticeable variations: both were severely dwarf and had crimped leaves and many malformed flowers. The tetraploid regenerants had several horticulturally attractive characteristics compared with the diploid controls, such as longer shoots, thicker stems, and larger flowers. Thus regeneration of tetraploid plants from 1-year-old embryogenic callus cultures offers a possibility to improve the horticultural value of T. hirta, although regeneration of trueness-to-type plants is essential for utilizing the cultures for micropropagation and genetic transformation.     

Changes in the gene expression of adiponectin and glucose transporter 12 (GLUT12) in lactating and non-lactating cows

Glucose delivery and uptake by the mammary gland is a rate‐limiting step in milk synthesis. Insulin resistance is believed to increase throughout the body following the onset of lactation. To study glucose metabolism in peak‐, late‐, and non‐lactating cows we analyzed the expression of an adipokine, namely, adiponectin, decreased insulin resistance, leptin, and a novel insulin‐responsive glucose transporter (GLUT12) in the adipose tissue and mammary gland by using real‐time polymerase chain reaction. Our results demonstrated that the mRNA level of adiponectin in the adipose tissue was greater in non‐lactating cows than in peak‐lactating cows. In the adipose tissue, there were no significant differences in the abundance of GLUT12 mRNA between the peak‐, late‐, and non‐lactating cows. In contrast, in the mammary gland, the mRNA level of GLUT12 was greater in non‐lactating cows than in peak‐ and late‐lactating cows. In the adipose tissue, the mRNA level of leptin and peroxisome proliferator‐activated receptor gamma 2 (PPARγ2) was greater in non‐lactating cows than in peak‐lactating cows. The results of the present study suggest that in lactating cows adiponectin plays an important role in insulin resistance in the adipose tissue; in the mammary gland, GLUT12 expression is believed to be an important factor for insulin‐dependent glucose metabolism.