Microbiota analysis of Caspian Sea yogurt, a ropy fermented milk circulated in Japan

We analyzed the microbiota of domestic ropy fermented milk, Caspian Sea yogurt (or 'kasupikai yohguruto' in Japanese), circulated in Japan. We collected six varieties from five localities. Lactococcus ( L) lactis ssp. cremoris was isolated from all samples as the dominant strain at levels of 10⁸−10⁹ CFU/g. We show this strain produces an extracellular polysaccharide (EPS) that causes the unique characteristic viscosity of the product. From analysis of the RAPD pattern of 60 bacterial isolates from the six samples, we found that 59 strains from a total of 60 isolates were identical and produced this viscosity. Furthermore, PFGE analysis of representative strains from each sample indicated that the isolates could be classified into four subgroups. This suggests these L. lactis ssp. cremoris strains found in Caspian Sea yogurt may have been slightly mutated during subculture in Japan. In addition, Lactobacillus (L.) sakei ssp. sakei were isolated from three samples; L. plantarum , Gluconoacetobacter sacchari and Acetobacter aceti were isolated from two samples; and L. paracasei , L. kefiri , Leuconostoc ( Leu. ) mesenteroides were isolated from one sample.     

Enzymatic and mechanistic studies on the formation of N-phenylglycolohydroxamic acid from nitrosobenzene and pyruvate in spinach leaf homogenate

The biotransformation mechanism of an unknown metabolite formed enzymatically from nitrosobenzene (NOB) and pyruvate in spinach (Spinacea oleracea L.) was investigated using spinach leaf homogenate. The unknown metabolite was identified as N-phenylglycolohydroxamic acid (PGA). The activity of PGA formation was decreased by l-alanine, increased by l-serine, and completely inhibited by aminooxyacetic acid, an inhibitor of transaminases. These results indicate that the transaminase participates in PGA formation. Indeed, hydroxypyruvate and alanine were produced in the transamination between pyruvate and serine. Hydroxypyruvate served as a direct-acting glycoloyl donor for PGA formation. A good correlation between the activities of the 200 g supernatant of spinach homogenate and commercial yeast transketolase for PGA formation from several glycoloyl donors was obtained. These results suggest the following mechanism for PGA formation from NOB and pyruvate: transamination of l-serine into hydroxypyruvate, which serves as a glycoloyl donor to NOB.     

Metabolite profiling of shoot extract,root extract,and root exudate of rice under nitrogen and phosphorus deficiency

ABSTRACT

Root exudate is derived from plant metabolites and its composition is affected by plant nutrient status. A deficiency of mineral nutrients, such as nitrogen (N) and phosphorus (P), strongly affects the type and amount of plant metabolites. We applied a metabolite profiling technique to investigate root exudates of rice plants under N and P deficiency. Oryza sativa was grown in culture solution containing two N levels (0 and 60 mg N L^?1) or two P levels (0 and 8 mg P L^?1). Shoot extracts, root extracts, and root exudates were obtained from the rice plants 5 and 15 days after transplanting and their metabolites were determined by capillary electrophoresis/time-of-flight mass spectrometry. Shoot N concentration and dry weight of rice plants grown at ?N level were lower than those of plants grown at +N level. Shoot P concentration and dry weight of rice plants grown at ?P level were lower than those of plants grown at +P level. One hundred and thirty-two, 127, and 98 metabolites were identified in shoot extracts, root extracts, and root exudates, respectively, at the two N levels. One hundred and thirty-two, 128, and 99 metabolites were identified in shoot extracts, root extracts, and root exudates, respectively, at the two P levels. Seventy-seven percent of the metabolites were exuded to the rhizosphere. The concentrations of betaine, gamma-aminobutyric acid, and glutarate in root exudates were higher at both ?N and ?P levels than at their respective high levels. The concentration of spermidine in root exudates was lower at both ?N and ?P levels than at their respective high levels. The concentrations of the other metabolites in root exudates were affected differently by plant N or P status. These results suggest that rice roots actively release many metabolites in response to N and P deficiency.     

Solubilization of Insoluble Inorganic Phosphate by Hyphal Exudates of Arbuscular Mycorrhizal Fungi

Increased phosphate (P) uptake in plants by arbuscular mycorrhizal (AM) fungi is thought to depend mainly on the extension of external hyphae into soil. On the other hand, it is known that the hyphae of some kinds of ectomycorrhizal fungi release organic acids into soil and that they dissolve the insoluble inorganic P. This study collected hyphal exudates of AM fungi within compartmentalized pot culture and clarified their ability to solubilize insoluble inorganic P. Sterilized Andisol was packed in pots that were separated into root and hyphal compartments with a nylon net of 30 μm pore size. Seedlings of Allium cepa inoculated with AM fungi, Gigaspora margarita, or Glomus etunicatum were grown. Control pots were not inoculated. Mullite ceramic tubes were buried in the soil of each compartment and soil solution was collected. The anionic fraction of the soil solution was incubated with iron phosphate (4 mg FePO₄ in 1 mL of 0.4 acetate buffer). Solubilized P was measured. The AM colonization of plants inoculated with G. margarita and G. etunicatum was 86% and 54%, respectively. Adhesion of external hyphae was observed on the surface of the mullite ceramic tubes buried in soil of the hyphal compartment. Colonization of both fungi increased shoot P uptake and growth. Soil solution collected from the hyphal compartments of both fungi solubilized more P than did that from uninoculated plants. It is suggested that hyphal exudates can contribute to increased P uptake of colonized plants.     

Comparative toxicity of Al3+, Yb3+, and La3+ to root-tip cells differing in tolerance to high Al3+ in terms of ionic potentials of dehydrated trivalent cations

Abstract

Green manure legumes are often used to compare biomass production as well as nitrogen-fixing capacity. Mineral deficiency often limits the symbiotic nitrogen fixation of many legumes, thus limiting their productivity despite their high yielding potential (O’Hara et al. 1988; Flis et al. 1993). Leguminous species require large amounts of P for growth, nodulation, and nitrogen fixation. Consequently, they are often unable to grow in acid soils with low available P. The low P availability in tropical acid soils often arises from fixation of P by Al and Fe in soil. Generally, Al and Fe-phosphates are relatively unavailable to plants (McLachlan 1976; Ae et al. 1990).     

The plasma membrane intactness of root-tip cells is a primary factor for Al-tolerance in cultivars of five species

We investigated the role of the cell wall and plasma membrane (PM) of root-tip cells in Al tolerance in Al-tolerant and Al-sensitive cultivars of five plant species (rice, maize, pea, wheat, and sorghum). No correlation was found between the differences in Al tolerance and the cation exchange capacity of cell walls isolated from root-tips (0–1 em). Preliminary exposure to Al for 1 h was sufficient to inhibit subsequent root re-elongation in an Al-free solution, and the inhibitory effect was more pronounced in the Al-sensitive cultivars than in the Al-tolerant ones. Together with the inhibition of root re-elongation, the PM of the root-tip cells of all the Al-sensitive cultivars was more permeabilized than that of the Al-tolerant cultivars, based on the FDA-PI fluorescence staining technique. Exposure for 30 min to Al treatment at 100 µM significantly increased the PM permeability of protoplasts isolated from the root-tips for the Al-sensitive pea cultivar placed in a moderately hypotonic medium. Protoplasts from root-tip portions of all the Al-sensitive cultivars took up more Al than those of the Al-tolerant ones when treated with 100 pM Al under isotonic conditions for 30 min. The co-existence of DNP or hypotonic conditions led to a larger increase of Al uptake by the protoplasts from Al-sensitive maize cultivars. These results suggest that Al ions rapidly alter the PM of the root-tip portion in the Al-sensitive cultivars, irrespective of plant species, resulting in an increase of the PM permeability.     

Genetic variation in height growth curves observed in three clonal tests of sugi (Cryptomeria japonica D. Don) in Kyushu

Genetic variation in the height growth curves was investigated by analyzing data from three clonal trials of sugi (Crytomeria japonica D. Don) in Kyushu. Average height of twelve clones measured at each of five years until 30 years of age were fitted to the Chapman-Richards function with two parameters model. Clonal variation in the two parameters (an upper asymptote and a rate of growth parameter) was statistically significant across the tests, indicating that height growth pattern is under genetic control. The type of growth curve was clearly classified with the two parameters and agreed with the empirical classifications of the growth habit of local varieties. The rate of growth parameter was correlated with total height up to 20 years of age, while the upper asymptote was correlated only with the last ten years’ increment from 21 to 30 years where the clonal repeatability was decreasing. This indicates that a clonal selection for long rotation management system would not be reliable unless the height is measured accurately with more advanced devices or it is adjusted by the current increment of diameter.     

Identification of oyster-derived hypotensive peptide acting as angiotensin-I-converting enzyme inhibitor

Angiotensin-I-converting enzyme (ACE) plays a crucial role in the crisis of hypertension. Some peptides that originate from protease hydrolysates are known to suppress ACE activity in vitro and in vivo. Here, we investigated whether trypsin hydrolysate of oyster Crassostrea gigas showed hypotensive activity and ACE inhibition. The hydrolysate significantly suppressed systolic blood pressure and ACE activity in spontaneously hypertensive rats following a one-shot oral administration and a long-term feeding experiment lasting 9 weeks. Each hydrolysate from oyster tissue showed ACE inhibitory activity, indicating the hypotensive effect was due to synergism. One potent ACE inhibitory peptide, Asp-Leu-Thr-Asp-Tyr, was identified from the hydrolysate of the striate muscle, and the peptide exhibited hypotensive activity in vivo. Protease digestion analysis suggested that Asp-Tyr could be the real effector of this penta-peptide in vivo.     

Identification of a potent immunostimulatory oligodeoxynucleotide from Streptococcus thermophilus lacZ

Immunostimulatory sequences of oligodeoxynucleotides (ODNs), such as CpG ODNs, are potent stimulators of innate immunity. Here, we identified a strong immunostimulatory CpG ODN, which we named MsST, from the lac Z gene of Streptococcus (S.) thermophilus ATCC19258, and we evaluated its immune functions. In in vitro studies, MsST had a similar ability as the murine prototype CpG ODN 1555 to induce inflammatory cytokine production and cell proliferation. In mouse splenocytes, MsST increased the number of CD80+CD11c+and CD86+CD11c+ dendritic cells and CD4+CD25+ regulatory T cells. We also analyzed the effects of MsST on the expression of regulatory cytokines by real-time quantitative PCR. MsST was more potent at inducing interleukin-10 expression than the ODN control 1612, indicating that MsST can augment the regulatory T cell response via Toll-like receptor 9, which plays an important role in suppressing T helper type 2 responses. These results suggest that S. thermophilus , whose genes include a strong Immunostimulatory sequence-ODN, is a good candidate for a starter culture to develop new physiologically functional foods and feeds.