Distribution of the FoToml gene encoding tomatinase in formae speciales of Fusarium oxysporum and identification of a novel tomatinase from F. oxysporum f. sp. radicis-lycopersici, the causal agent of Fusarium crown and root rot of tomato

The antifungal glycoalkaloid -tomatine accumulates in tomato plants and may protect plants from fungal infection. Fusarium oxysporum f. sp. lycopersici, the causal agent of vascular wilt of tomato, produces a tomatinase (FoToml) that degrades -tomatine to the nontoxic compounds tetrasaccharide lycotetraose and tomatidine. Induction of tomatinases and the distribution of FoToml homologs were examined among 30 strains belonging to 16 formae speciales of F. oxysporum. Tomatinase activity was found in 27 strains belonging to 15 formae speciales, but FoToml homologs (>98% sequence identity) were detected in only six strains belonging to four formae speciales. To identify tomatinases other than FoToml, -tomatine-inducible proteins of another tomato pathogen F. oxysporum f. sp. radicis-lycopersici were analyzed by two-dimensional gel electrophoresis. A protein with a molecular mass of 64kDa accumulated in the -tomatine-induced culture filtrates, and the protein had tomatinase activity, degrading -tomatine to lycotetraose and tomatidine.     

Permeation of carboxylates as ionic compounds into fish meats by soaking and its osmotic dehydrating effect

Permeability of Na-gluconate, Na-malate and Na-acetate into fish-meat strips by soaking, and their osmotic dehydrating effects were investigated in comparison with those of sodium chloride (NaCl) and sorbitol. Carboxylic acids, anion species of carboxylates, were less permeable into soaked meats than chlorine of NaCl, and their permeability was similar to that of sorbitol. However, the permeation of sodium, cation species of the carboxylates and NaCl, was promoted with the concentration of free sodium ion dissociated in soaking solutions. The ratio of sodium to the anion species in soaked meats was varied, depending on the numbers of the sodium atom in the molecules as well as the dissociation degree of the ionic compounds in the soaking solution. Furthermore, the osmotic dehydrating effects of Na-gluconate and Na-malate were analogous with that of sorbitol, and higher than those of NaCl and Na-acetate. These results indicated that the permeation characteristics and the osmotic dehydrating effects of the carboxylates were associated with their ionization in soaking solution as well as their molecular weights.     

Hydathode function and changes in contents of elements in eddo exposed to zinc in hydroponic solution

To explore the zinc stress response in eddo, plants were grown for 15 d in hydroponic solutions containing 1 (control), 200, and 1000 μM zinc, and the hydathode function and changes in the contents of various elements in these plants were investigated. Under 1000 μM zinc, the dry weights of leaf blades and roots are decreased by 17 and 42%, respectively. The zinc contents in leaf blades, petioles, corms, and roots increased with increasing zinc levels in the solution. The zinc content in roots was 6.57 mg g^?1 dry weight, which was 2.8–4.3 times higher than in other plant parts under 1000 μM zinc. These results demonstrate that the severe root damage occurring under 1000 μM zinc is due to higher zinc content in the roots. Under zinc stress, the contents of iron and copper in roots increased, but the contents of magnesium and calcium in petioles, corms, and roots, iron in leaf blades and corms, and manganese in petioles and corms decreased. In the guttation fluid, the concentrations of zinc, magnesium, and potassium increased, while the iron concentration decreased under 1000 μM zinc. Thus, elemental changes occurred in the guttation fluid as well as in different plant parts in eddo. In the 200 and 1000 μM zinc treatments, the amount of zinc eliminated via guttation was 2.8 and 8.5 times higher, respectively, than in the control. The results indicate that guttation partly contributes to the excretion of excess zinc under zinc stress conditions.     

Analysis of plasmids encoding the tyrosine decarboxylase gene in Tetragenococcus halophilus isolated from fish sauce

In order to elucidate mechanisms of tyramine accumulation during fish sauce production, two tyramine-producing bacterial strains, referred to as TyrA and TyrB, were isolated from fish sauce mash accumulating over 141 mg of tyramine per 100 g of sample. Both strains were identified as Tetragenococcus halophilus based on phenotypic characterization and a 16S rRNA gene sequence analysis. Molecular analysis of the tyramine-producing gene in the two strains confirmed the presence of a ~30-kb plasmid encoding a single copy of the pyridoxal phosphate-dependent tyrosine decarboxylase gene (tdcA) along with three other genes related to tyramine synthesis (tdc cluster). The complete nucleotide sequences of plasmids extracted from the two strains indicated that both plasmids were almost identical, except for a 1.6-kb transposon sequence in the plasmid from the strain TyrB. Both plasmids had a replication region, a plasmid maintenance region, and two putative mobile genetic elements located upstream and downstream of the tdc cluster. This structure was identical to that of tetragenococcal plasmids encoding histidine decarboxylase (hdcA), which were sequenced previously. These results suggest a common origin for plasmids encoding hdcA and tdcA. In addition, the genes for both these biogenic amines are distributed among tetragenococcal species via this plasmid.     

Chemical Characteristics of Fog Water at Mt. Tateyama, Near the Coast of the Japan Sea in Central Japan

Measurements of the chemical composition of fog water at Murododaira (altitude, 2,450 m), on the western slope of Mt. Tateyama near the coast of the Japan Sea, were performed each autumn from 2004 through 2007. Strong acidic fogs (pH?<?4) containing high concentrations of nssSO ₄ ^2? were frequently observed in the autumn of 2005, when the air mass at Mt. Tateyama originated mainly from the polluted regions of Asia. The ratio of NO ₃ ^? /nssSO ₄ ^2? in fog water was relatively high in 2004 and 2007. High concentrations of nssCa²⁺ derived from dust particles were detected in 2006. Background Kosa particles might have been predominant in the free troposphere and could have neutralized acidic fogs in the autumn of 2006. High concentrations of sea-salt components were also observed in October 2005. The sea-salt particles might have been transported from the Pacific Ocean by a strong typhoon, and significant Mg²⁺ loss was observed. Peroxides higher than 100 μM, which are seriously harmful to vegetation were sometimes detected.     

Phospholipase Cζ (PLCζ) versus postacrosomal sheath WW domain‐binding protein (PAWP): Which molecule will survive as a sperm factor?

During mammalian fertilization, sperm is fused with the oocyte's membrane, triggering the resumption of meiosis from the metaphase II arrest, the extrusion of the second polar body, and the exocytosis of cortical granules; these events are collectively called 'oocyte activation.' In all species studied to date, the transient rise in the cytosolic level of calcium (in particular, the repeated calcium increases called 'calcium oscillations' in mammals) is required for these events. Researchers have focused on identifying the factor(s) that can induce calcium oscillations during fertilization. Sperm‐specific phospholipase C, i.e., PLC zeta (PLCζ), is a strong candidate of the factor(s), and several research groups using different species obtained evidence that PLCζ is a sperm factor that can induce calcium oscillations during fertilization. However, postacrosomal sheath Tryptophan‐Tryptophan (WW)—domain‐binding protein (PAWP) was recently shown to have a pivotal role in inducing calcium oscillations in some species. In this review, we focus on PLCζ and PAWP as sperm factors, and we discuss this controversy: Which of these two molecules survives as a sperm factor?     

Innate immune response of mammary gland induced by intramammary infusion of Bifidobacterium breve in lactating dairy cows

This study aimed to evaluate innate immune responses of mammary glands induced by intramammary infusion of Bifidobacterium breve in dairy cows. Somatic cell counts in quarters of cows showed a marked increase following B. breve infusion on days 1 and 2. Opsonized-stimulated chemiluminescence response in quarter milk was significantly (P<0.05) increased by B. breve infusion on days 1 to 3 compared to that of pre-infusion. Lactoferrin concentrations in B. breve-infused quarter milk increased significantly (P<0.05) on days 2 to 4 and 6 compared to those of pre-infusion. IgG and IgA concentrations in B. breve-infused quarters significantly (P<0.05) increased on days 2 to 4 for IgG and days 3, 4, 6 and 8 for IgA compared to those of pre-infusion. Interleukin (IL)-1β and IL-8 mRNA levels in somatic cells from B. breve-infused quarters were significantly (P<0.05) upregulated on day 1 compared to those on days 0 and 14. Conversely, IL-6 mRNA levels in somatic cells from B. breve-infused quarters on days 0, 1 and 14 and NF-κB mRNA levels on day 0 were significantly (P<0.05) down-regulated compared to those of control. IL-1β, tumor necrosis factor (TNF)-α and IL-6 concentrations increased on days 1, 3 and 7 after B. breve infusion in quarters. Intramammary infusion of B. breve (3 × 10⁹ cfu) induces a massive influx of leukocytes and enhances innate immune response in mammary glands. This event may contribute to the enhancing host defense in the mammary gland.     

Fetal and neonatal goiter in cynomolgus monkeys following administration of the antithyroid drug thiamazole at high doses to dams during pregnancy

To evaluate morphologic alterations in the thyroid gland in the second generation in cynomolgus monkeys, pregnant dams were exposed to high doses of thiamazole. In Experiment A, dams received thiamazole intragastrically via a nasogastric catheter from gestation day (GD) 50 to GD 150 or on the day before delivery. Initially, the dose level was 20 mg/kg/day (10 mg/kg twice daily); however, the dose level was subsequently decreased to 5 mg/kg/day (2.5 mg/kg twice daily), since deteriorated general conditions were observed in two dams. Six out of seven neonates died on the day of birth. The cause of neonatal death was tracheal compression and suffocation from goiter. The transplacental exposure to thiamazole affected the fetal thyroid glands and induced goiter in all neonates. The surviving neonate was necropsied 767 days after discontinuation of thiamazole exposure and showed reversibility of the induced changes. In Experiment B, dams were intragastrically administered thiamazole at 5 mg/kg/day (2.5 mg/kg twice daily) for treatment periods from GDs 51 to 70, 71 to 90, 91 to 110, 111 to 130 and 131 to 150. All fetuses showed enlarged thyroid glands but were viable. Histopathologically, hypertrophy and/or hyperplastic appearance of the follicular epithelium of the thyroid gland was observed at the end of each treatment period. The most active appearance of the follicular epithelium, consisting of crowded pedunculated structure, was demonstrated at end of the treatment period from GD 131 to 150. This is the first report on the morphology of fetal and neonatal goiter in the cynomolgus monkey.     

不同种植密度下温室间作玉米的水分利用情况与其营养元素吸收特征的关系

Dent corn, as a catch crop used for salt removal, was cultivated at different densities, i.e., 7.3 （low density）, 59.7 （normal density）, and 119.5 plants m-2 （high density）, during a 50 d fallow period after cultivation of a commercial crop in a greenhouse, to analyze the characteristics of nutrient salt （N, K, Mg, and Ca） uptake by roots and to study the effect of plant density on the characteristics associated with crop water use. Leaf area index for the high and normal density treatments reached extremely high values of 24.3 and 14.9, respectively. These values induced higher transpiration rates that were estimated using the Penman-Monteith model with the incorporation of specific parameters for crop and greenhouse conditions. The total N, K, Mg, and Ca contents in the crop canopy at harvest were 26.8, 13.0, 1.0, and 1.7 g m-S, respectively, under the high density treatment. The dynamics of salt uptake rates for high, normal, and low density treatments were evaluated by assessing weekly changes in salt content, and were subsequently compared against the transpiration rate. A positive linear relationship was obtained between these 2 parameters for all 3 density treatments and all tested salts. Hence, higher transpiration rates caused higher salt uptake rates through water absorption. On the other hand, salt uptake efficiency per unit water use by cultivation was lower in the low density treatment. Therefore, management procedures with dense planting that induce higher transpiration rates and lower evaporation rate are extremely important for the effective cultivation of corn catch crops.     

Synthesis of citrulline from ornithine by the small intestinal mucosa of cattle

Three segments of cattle small intestine (duodenum, upper jejuno‐ileum and lower jejuno‐ileum) were examined in an in vitro system for activity of ornithine carbamoyltransferase (OCT; EC 2.1.3.3) which is involved in the synthesis of citrulline (Cit) from ornithine (Orn). The mucosa of the three segments of small intestine was collected from Japanese black cattle, homogenized and then centrifuged. The supernatant fraction was used as the crude OCT enzyme solution. The OCT activity was assayed by the production of Cit from Orn determined directly by HPLC. The optimal pH and temperature for OCT activities in the duodenum, upper jejuno‐ileum and lower jejuno‐ileum of cattle small intestine were 7.47 and 39°C. Little difference was observed between the three segments. The OCT activity in cattle kidney was also examined for comparison, and almost no activity was found. The OCT activities in crude enzyme solutions of the three segments of small intestine were stable for up to one month of storage at ?20°C in Tris HCl buffer solution. Finally, the role of the small intestine in supplying Cit as a precursor for arginine synthesis in cattle kidney was discussed.