Flagellin glycosylation is ubiquitous in a broad range of phytopathogenic bacteria

Glycosylation of flagellin is known to be involved in filament stabilization, motility, and virulence in Pseudomonas syringae. Here we investigated flagellin glycosylation in other phytopathogenic bacteria. Analyses of deduced amino acid sequences, glycostaining, and molecular masses of purified flagellins revealed that flagellins from all phytopathogenic bacteria investigated were glycosylated. Furthermore, the flagellin in a glycosylation-defective mutant of Xanthomonas campestris pv. campestris (Xcc) had a reduced molecular mass, and motility and virulence of the mutant toward host leaves decreased. These results suggest that flagellin glycosylation is ubiquitous in most phytopathogenic bacteria and that flagellin glycosylation is required for virulence in Xcc.     

Upregulation of the genes for ferritin,RNase, and DnaJ in leaves of rice plants in response to sulfur deficiency

Microarray analysis was carried out with mRNAs isolated from aerial portions of rice plants subjected to sulfur deficiency. Among the candidate clones regulated by sulfur deficiency, three genes were identified by northern analysis that showed altered levels of mRNA accumulation. Genes similar to those for ferritin, S-like ribonuclease, and DnaJ-like protein were down-, down-, and upregulated by sulfate deficiency, respectively. These genes were found for the first time to be responsive to sulfur nutrition in the present study.     

Investigation on hydrogen abstraction from methyl glucoside by active oxygen species under oxygen delignification conditions III: effects of the origin of active oxygen species

Carbohydrate model compounds methyl β-d-glucopyranoside (MGPβ), methyl α-d-glucopyranoside (MGPα), and methyl β-d-mannopyranoside (MMPβ) and the deuterium compounds of MGPβ labeled at the anomeric or C-2 positions (MGPβ-1D, MGPβ-2D) were reacted with active oxygen species (AOS) generated in situ by reactions between O₂ and a co-treated phenolic lignin model compound, 4-hydroxy-3-methoxybenzyl alcohol (VAlc), under conditions simulating oxygen delignification (0.5 mol/l NaOH, 0.36 mmol/l Fe³⁺, 1.1 MPa O₂, 95°C). MGPβ was degraded more than MGPα but less than MMPβ when the pairs MGPβ/MGPα and MGPβ/MMPβ, respectively, were treated, which indicates that the configurational differences at the anomeric and C-2 positions influence the reactivity of AOS toward these compounds. When the pairs MGPβ/MGPβ-1D and MGPβ/MGPβ-2D were treated, no clear kinetic isotope effects were observed in either case. These results contrasted with those obtained when another phenolic compound, 2,4,6-trimethylphenol (TMPh), was used as the AOS generator instead of VAlc under exactly the same conditions. Clear kinetic isotope effects were observed when using TMPh. Because it is not easily accepted that the anomeric and C-2 hydrogen abstractions are minor reaction modes only for AOS generated in the VAlc system, it is suspected that the AOS do not show any clear kinetic isotope effect even though the AOS abstract an objective hydrogen.     

Leaf margin phenotype-specific restriction-site-associated DNA-derived markers for pineapple (Ananas comosus L.)

To explore genome-wide DNA polymorphisms and identify DNA markers for leaf margin phenotypes, a restriction-site-associated DNA sequencing analysis was employed to analyze three bulked DNAs of F1 progeny from a cross between a ‘piping-leaf-type’ cultivar, ‘Yugafu’, and a ‘spiny-tip-leaf-type’ variety, ‘Yonekura’. The parents were both Ananas comosus var. comosus. From the analysis, piping-leaf and spiny-tip-leaf gene-specific restriction-site-associated DNA sequencing tags were obtained and designated as PLSTs and STLSTs, respectively. The five PLSTs and two STSLTs were successfully converted to cleaved amplified polymorphic sequence (CAPS) or simple sequence repeat (SSR) markers using the sequence differences between alleles. Based on the genotyping of the F1 with two SSR and three CAPS markers, the five PLST markers were mapped in the vicinity of the P locus, with the closest marker, PLST1_SSR, being located 1.5 cM from the P locus. The two CAPS markers from STLST1 and STLST3 perfectly assessed the ‘spiny-leaf type’ as homozygotes of the recessive s allele of the S gene. The recombination value between the S locus and STLST loci was 2.4, and STLSTs were located 2.2 cM from the S locus. SSR and CAPS markers are applicable to marker-assisted selection of leaf margin phenotypes in pineapple breeding.     

Search for an index for the taste of Japanese Black cattle beef by panel testing and chemical composition analysis

To search for an index for chemical composition related to superior taste in Japanese Black beef, we conducted panel tests and analyzed the chemical composition of seven beef brands. Thirty‐five sirloin beefs from five heifers were used in this study, sold under seven beef brands graded as more than A4 on the Japanese Meat Grade scale. The chemical composition analyses assessed both raw and roasted meat, the latter of which was roasted under the same conditions as those used for the panel test. Results of the panel test and chemical composition analyses revealed that fatty acid composition, sugar content, adenosine triphosphage (ATP)‐related compounds, amino acid composition and odor composition in the sirloin meat differed among beef brands. Furthermore, the correlations of chemical compositions between roasted and raw meat were significantly high. Sugar content and ATP‐related compounds in roasted meat were significantly correlated with the item ‘overall evaluation’ of the panel test. ATP‐related compounds, such as inosinic acid, carnosine and taurine, in roasted and raw meat were correlated significantly with the item ‘umami intensity’ of the panel test. These results suggest that the composition of these components is important for an index related to the overall evaluation of beef.     

Histopathological characterization of the skeletal myopathy in rasH2 mice carrying human prototype c-Ha-ras gene

A skeletal myopathy is found in approximately 100% of rasH2 mice. To confirm detailed features of the rasH2 skeletal myopathy, the biceps femoris, diaphragm, triceps brachii, gastrocnemial (types I and II fiber-mixed muscles) and soleus muscle (type I fiber-dominant muscle) obtained from male rasH2 and non-transgenic littermates aged 10-13 and 34 weeks were examined. Variations in the muscle fiber size, early-scattered degeneration/necrosis and regeneration of muscle fibers were detected in 10-13-week-old rasH2 mice. The severity of the above muscular lesions was more prominent in older rasH2 mice. These lesions were noted in the type II myofiber dominant muscles (biceps femoris, triceps brachii and gastrocnemial). NADH-TR stain clearly demonstrated a disorganized intermyofibrillar network and necrotic change in muscle fibers. No specific morphological changes, like rod structure or tubular aggregation seen in some types of myopathy, were noted in Gomori trichrome and NADH-TR stains in the rasH2 mouse like in many types of muscular dystrophy. Electronmicroscopically, occasional muscle fiber degeneration/regeneration, invaded phagocytic cells, indistinct Z-band suggesting excessive contraction and dilatation of the sarcoplasmic reticulum were observed. In summary, the skeletal myopathy occurring in rasH2 mice is consistent with muscular dystrophy characterized morphologically by progressive degeneration and regeneration of myofibers. The myopathy is confined to the type II myofiber predominant muscles and is not associated with any pathognomonic lesions. These characteristics will provide us with a useful model for research in muscular dystrophy of diverse myofibers.     

The absence of Rhodococcus equi in Mongolian horses

In native Mongolian horses, the incidence and distribution of Rhodococcus equi are poorly understood. One hundred and fourteen equine fecal samples and 71 soil samples were collected from the camp sites of 26 nomadic families located in three areas less than 100 km from Ulaanbaatar, Mongolia. Five fecal samples were also collected from foals of Przewalski's Horses introduced into the Hustai National Park, Mongolia. No R. equi was isolated from the Mongolian horses or the soil samples. However, three colonies of R. equi were isolated from two fecal samples collected from foals of Przewalski's Horses. These isolates were avirulent, with neither 15- to 17-kDa antigens (VapA) nor a 20-kDa antigen (VapB) genes being detected. We concluded that native Mongolian horses and their environment appear free from contamination with R. equi.     

Effect of fat content on sensory characteristics of marbled beef from Japanese Black steers

To analyze the sensory characteristics of meat samples with a crude fat content between 23.8% and 48.6% taken from 34 Japanese Black steers, we grilled the meat and subjected it to analytical sensory evaluation. We also measured the amounts of moisture, protein, nucleic acid and glutamic acid. An increase in crude fat content increased the tenderness, juiciness, and fattiness in the meat quality evaluation. An increase in crude fat content reduced the crude protein and moisture contents; it also slightly reduced the nucleic acid and glutamic acid contents, although when the reductions in these umami components were assessed relative to the moisture content they changed little. Increasing the fat content up to a certain point greatly enhanced the umami intensity and beef flavor intensity in the meat quality evaluation and raised the overall evaluation score; the peak of the appropriate crude fat content for these purposes was about 36%.     

Inorganic and organic osmolytes accumulation in five halophytes growing in saline habitats around the Aiding Lake area in Turpan Basin,Northwest China

Halophytes dominate the plant community in saline soils. Here, osmoregulation via the accumulation of osmolytes is the basic strategy by which plants survive salinity stress. We investigated the accumulation of inorganic and organic osmolytes in the leaves of five halophytes (Tamarix hispida, Halocnemum strobilaceum, Kalidium foliatum, Karelinia caspica, and Phragmites australis) growing in the dry lakebed of Aiding Lake, Xinjiang, China. The succulent euhalophytes (H. strobilaceum and K. foliatum) accumulated large amounts of Na⁺, whereas other species had low Na⁺ concentrations. P. australis contained high concentrations of soluble carbohydrates, mainly sucrose, and amino acids, such as proline and alanine. K. caspica accumulated large quantities of mannitol. H. strobilaceum and K. foliatum had high glycine betaine contents. Only T. hispida accumulated γ-butyro betaine, which was found in high concentrations. Our findings indicate that at least four types of osmolytes (carbohydrates, polyols, amino acids, and betaines) function either alone, or in combination in the osmoregulation of these halophytes.     

Immunostimulating activity of a crude polysaccharide derived from green tea (Camellia sinensis) extract

Green tea extract is well-known to reduce the risk of a variety of diseases. Here, we investigated the immunostimulating activity of tea polysaccharide (TPS), one of the main components in green tea extract. The water extracts from mature or immature tea leaves were precipitated by using ethanol at room temperature. The sediment was washed with ethanol and acetone alternately and then dried. We used the phagocytic activity of macrophage-like cells as an indicator of immune function activation. Chemical components were analyzed by HPLC. The immunostimulating activity of TPS from immature leaves extract was higher than that of TPS from mature leaves, and its activities were dependent on the content of strictinin in the leaf extract. Futhermore, a mixture of catechin and TPS that removed polyphenols did not increase the immunostimulating activity. These results suggest that the catechin-polysaccharide complex is a very important molecule in the immunomodulating activity of tea extracts.