Norsalsolinol uptake into secretory vesicles via vesicular monoamine transporter and its secretion by membrane depolarization or purinoceptor stimulation in PC12 cells.

The intracellular dynamics of norsalsolinol, a neurotoxin candidate causing parkinsonism-like symptoms, in PC12 cells was studied. We found that dopamine and norsalsolinol are co-localized to secretory granule layer by sucrose density gradient in norsalsolinol-treated PC12 cells. The norsalsolinol was actively taken up into isolated secretory vesicle fraction from PC12 cells with a Km value of 41.5+/-6.8 microM. The uptake of 10 microM of norsalsolinol was sensitive to reserpine (1 microM), an inhibitor of vesicular dopamine transporter, and dopamine, an endogenous substrate, but insensitive to GBR-12909, an inhibitor of dopamine transporter on plasma membrane. In norsalsolinol-treated PC12 cells, exposure to high K+ or ATP resulted in simultaneous release of norsalsolinol and dopamine. Time course of a release of dopamine and that of norsalsolinol evoked by 50 mM KCl or 100 microM ATP corresponded to each other. These releases were dependent on the concentrations of secretagogues. These data suggest that norsalsolinol is taken up with dopamine into secretory vesicle via vesicular catecholamine transporter.     

The neuroexcitatory,convulsive and lethal effects of lindane analogs on Periplaneta americana (L.)

Lindane analogs, in which one or two chlorine atoms are replaced by various substituents, exhibit convulsive and lethal effects on Periplaneta americana (L.). Some analogs are as active as lindane. These effects are well related to the neuroexcitatory action which produces after-discharges in the central nervous system. Carbamate insecticides have potent convulsive and lethal effects on P. americana, which are also related to after-discharges produced in nerve cords. Convulsion caused by lindane analogs and carbamates is solely attributable to their neuroexcitatory effects on the nervous system, in spite of the difference in their ultimate modes of action.     

Identification of the medicinal off-flavor compound formed from ascorbic acid and (E)-hex-2-enal

A test apple beverage made up of apple juice (20%), high-fructose corn syrup (11.5%), citric acid (0.43%), trisodium citrate (0.02%), apple-odor flavor (0.1%), and ascorbic acid (0.02%) was stored at 40 °C and then analyzed for the change of odor in the beverage. Although no thermoacidophilic bacteria (TAB) were detected, a medicinal off-flavor was perceived after the 8 weeks of storage. Model experiments on the ingredients of the test apple beverage revealed that the off-flavor compound had been formed by ascorbic acid and (E)-hex-2-enal. Synthesis and NMR (1H, 13C, HMQC, and HMBC) analyses identified the compound as 6-propylbenzofuran-7-ol. The odor quality, retention index (RI), and mass spectrum of synthetic 6-propylbenzofuran-7-ol were identical with those of the medicinal odor compound from the test apple beverage. Sensory evaluation revealed the recognition thresholds for medicinal odor were 31.4 ppb in water and 24.0 ppb in apple beverage, and the detection thresholds were 19.6 ppb in water and 8.6 ppb in apple beverage, respectively. The quantified concentration of 6-propylbenzofuran-7-ol formed in test apple beverage was 90 ppb, approximately. This concentration was well above the odor threshold, so it was concluded that the compound was the source of the medicinal off-flavor.     

Purification and characterization of polyphenol oxidase from banana (Musa sapientum L.) pulp

Polyphenol oxidase (EC 1.10.3.1, PPO) in the pulp of banana (Musa sapientum L.) was purified to 636-fold with a recovery of 3.0%, using dopamine as substrate. The purified enzyme exhibited a clear single band on polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate (SDS)-PAGE. The molecular weight of the enzyme was estimated to be about 41000 and 42000 by gel filtration and SDS-PAGE, respectively. The enzyme quickly oxidized dopamine, and its K(m) value for dopamine was 2.8 mM. The optimum pH was at 6.5, and the enzyme activity was stable in the range of pH 5-11 at 5 degrees C for 48 h. The enzyme had an optimum temperature of 30 degrees C and was stable even after a heat treatment at 70 degrees C for 30 min. The enzyme activity was completely inhibited by L-ascorbic acid, cysteine, sodium diethyldithiocarbamate, and potassium cyanide. Under a low buffer capacity, the enzyme was also strongly inhibited by citric acid and acetic acid at 10 mM.     

Effect of bensulfuron-methyl (a sulfonylurea herbicide) on the soil bacterial community of a paddy soil microcosm

The effect of bensulfuron-methyl (BSM) on a soil microbial community in a model paddy microcosm was studied. Total bacterial numbers in the overlying water and surface soil were monitored for 2 months after the application of BSM at the field rate and a ten-fold field rate. Pentachlorophenol (PCP) was used for comparison. Neither chemical affected the total bacterial numbers remarkably, either in the overlying water or in the surface soil. In contrast, the nitrification potential was significantly suppressed by the BSM application. The bacterial community structure, as evaluated by the denaturing gradient gel electrophoresis (DGGE) of PCR amplification products from bacterial 16S rDNA, was unaffected by the BSM treatments over 8 weeks in the surface soil, compared with the control (no pesticide). In contrast, the surface soil exposed to PCP at a ten-fold field rate showed different patterns from the controls at 4 weeks and 8 weeks after application. The DGGE patterns of the overlying water were much more variable than those of the surface soil in any treatments. Cluster analysis showed that the BSM plots were classified within the same group as the control at 1 week after application and that the BSM and PCP plots from 2 weeks onward after application were grouped differently from the control. Of 22 clones excised from the DGGE gels, 20 clones belonged to the Proteobacteria and two belonged to the Verrucomicrobia. It was considered that the impact of BSM on the overall microbial community (total numbers, community structure of soil) was negligible, although BSM had an impact on some specific functions of the soil microbial community (nitrification) and a part of the community (overlying water).     

Thermal behavior of beta-1 subunits in lignin: pyrolysis of 1,2-diarylpropane-1,3-diol-type lignin model compounds

1,2-Diarylpropane-1,3-diol-type lignin model compounds, 1,2-bis(4-hydroxy-3-methoxyphenyl)propane-1,3-diol (1) and 1-(3,4-diethoxyphenyl)-2-(4-methoxyphenyl)propane-1,3-diol (2), were pyrolyzed at 500 degrees C for 4 s to clarify the thermal behavior of beta-1 subunits in lignin. Products were monitored by gas chromatography/mass spectrometry. The cleavage of the Calpha-Cbeta bond to produce benzaldehydes such as 4-hydroxy-3-methoxybenzaldehyde (9) and phenylethanals as the counterparts such as 4-hydroxy-3-methoxyphenylethanal (10) occurred in pyrolyses of both 1 and 2. In pyrolysis of 1, an oxetane pathway leading to the formation of Z/E-stilbenes without the gamma-CH2OH group such as Z/E-4,4'-dihydroxy-3,3'-dimethoxystilbene (3) was predominant. In pyrolysis of 2, the oxetane pathway was minor, while pathways producing a dimer with a =CgammaH2 group by loss of water and a dimer with an alpha-carbonyl group were predominant. Pyrolysis of Japanese cedar wood provided 3 and 10 in approximately 0.8% and 0.6% yields, respectively, based on the Klason lignin content, while pyrolysis of a guaiacyl bulk dehydrogenation polymer gave them in a very small amount.     

Development and evaluation of pyramiding lines carrying early or late heading QTLs in the indica rice cultivar ‘IR64’

The heading date is an important trait for determining regional and climatic adaptability in rice. To expand the adaptability of the indica rice cultivar ‘IR64’, we pyramided multiple early or late heading quantitative trait locus (QTLs) in the ‘IR64’ genetic background by crossing previously developed near-isogenic lines (NILs) with a single QTL for early or late heading. The effects of pyramiding QTLs were observed in three different climatic zones of the Philippines, Madagascar, and Japan. The early heading pyramiding lines (PYLs) headed 6.2 to 12.8 days earlier than ‘IR64’ while the late heading PYLs headed 18.8 to 27.1 days later than ‘IR64’. The PYLs tended to produce low grain yield compared to ‘IR64’. The low yield was not improved by combining SPIKE, which is a QTL that increases the number of spikelets per panicle. Conversely, ‘IR64-PYL(7+10)’ carrying Hd5 and Hd1 headed earlier, produced more tillers, and more panicles per m² than ‘IR64’, and mitigated the yield decrease in early heading. These results suggest that the effects of pyramided QTLs on heading date were consistent across various environments and PYLs could be used to enhance the adaptation of ‘IR64’ in other rice growing environments.     

Low water exchange Gracilariopsis bailiniae Zhang & B.M. Xia culture in intensive milkfish culture effluents: effects of seaweed density on seaweed production and effluent treatment

This study tested the feasibility of a low-cost seaweed biofiltration system for pond-based aquaculture through an indoor-integrated fish-seaweed culture experiment using weekly nutrient supply regime and different seaweed stocking densities. The culture experiment was conducted in glass aquaria that were stocked with Gracilariopsis bailiniae at 3 densities (low = 0.5 kg m^?2, middle = 2 kg m^?2 and high = 3.5 kg m^?2) and provided either with effluents from intensive milkfish (Chanos chanos) culture or with effluent-free seawater (control) as nutrient source. Stocking density was used as a factor in optimizing nutrient availability for growth and nutrient removal under such low water exchange conditions. Our results showed that G. bailiniae cultivated in milkfish effluents had higher growth, biomass and nitrogen yields than those cultivated in effluent-free seawater. Among the different stocking densities tested, highest growth rate (1.03 % day^?1) was obtained in the middle density. Increasing biomass and nitrogen yields were also obtained at this density until the end of the culture period. Poorer growth rates at low and high stocking densities were attributed to light limitation from phytoplankton and self-shading, respectively. Due to seaweed treatment, average outflow concentration of NH₄ ⁺ was reduced from half of its pretreated level. This study showed that a weekly effluent supply at 2 kg m^?2 seaweed stocking density can sustainably support the growth of G. bailiniae as long as the dissolved nutrients are present at high levels.     

Regulation of root-to-leaf Na and Cl transport and its association with photosynthetic activity in salt-tolerant soybean genotypes

ABSTRACT

Soil salinity is a major constraint to sustainable crop production. Genetic improvements are needed for growing soybean in salinity-prone environments. Salt-tolerant soybean genotypes alleviate a reduction in photosynthesis and growth under saline conditions; however, the detailed mechanisms involved remain unclear. Here, we aimed to clarify how Na and Cl root-to-leaf transport is quantitatively regulated, and to identify whether photosynthetic tolerance depends on traits associated with either stomata or with mesophyll tissues. Two pairs of pot-grown soybean near-isogenic lines (NILs) consisting of tolerant and susceptible counterparts, derived from a cross between salt-tolerant FT-Abyara and salt-sensitive C01, were subjected to salinity treatment in a rainout greenhouse. Comparison of photosynthetic responses between genotypes indicated that genotypic differences in salinity tolerance depended on the ability for sustained CO₂ assimilation in mesophyll tissues, rather than stomatal conductance. The ratio of photosynthetic rate to intercellular CO₂ concentration (A/Ci) declined exponentially with increasing Na and Cl concentration regardless of genotype, but tolerant genotypes effectively kept both elements at significantly low levels. Under saline conditions, tolerant genotypes reduced Na and Cl content at the two transport pathways: from root to stem, and from stem to leaf, but the reduction of Cl at each pathway was only minor. These results suggest that integrating genetic capacity for Cl transport regulation and osmotic adjustment should be an important target in salinity-tolerance soybean breeding.