Distribution of <Emphasis Type="Italic">tomato chlorotic dwarf viroid</Emphasis> in floral organs of tomato

In situ hybridization was used to analyze the distribution pattern of Tomato chlorotic dwarf viroid (TCDVd) in floral organs of tomato plants. Following TCDVd invasion of floral organs, it became localized only in sepals at an early developmental stage, then reached other floral organs at the flower opening stage, with the exception of part of the placenta and ovules. When distribution of TCDVd was compared with that of Potato spindle tuber viroid (PSTVd), TCDVd was not detected in the outer integument around the embryo sac even though PSTVd was able to invade there, suggesting that such specific distribution might reflect the frequent occurrence of viroid disease on crops caused by PSTVd-seed transmission.     

Isolation and characterization of aminopeptidase (Jc-peptidase) from Japanese cedar pollen (Cryptomeria japonica)

An aminopeptidase, Jc-peptidase, was purified from Japanese cedar pollen by seven steps, including precipitation with ammonium sulfate, ion-exchange chromatography, gel filtration, hydrophobic interaction chromatography on phenyl-agarose, and high-performance liquid chromatography. Purified Jc-peptidease has a molecular weight of 42 kDa and hydrolyzes the synthetic substrates of L-phenylalanyl-4-methylcoumaryl-7-amide (Phe-MCA) with Km = 5 x 10(-5) M, Tyr-MCA with Km = 7 x 10(-4) M, Leu-MCA with Km = 1 x 10(-3) M, and Met-MCA with Km = 1 x 10(-3) M. Other MCA analogues such as Arg-MCA or Glu-MCA failed to serve as its substrates. The activity was inhibited in the presence of phebestin, [(2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl-L-valyl]-L-phenylalanine, with Ki = 4.7 x 10(-5) M, or bestatin, [(2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl]-L-leucine, with Ki = 1.1 x 10(-4) M. According to amino acid sequence analysis, the N-terminal amino group seems to be blocked. The physiological function of the aminopeptidase (Jc-peptidase) has not been clarified in vivo.     

Effect of intermittent drainage in reduction of methane emission from paddy soils in Hokkaido,northern Japan

ABSTRACT

Emission of methane (CH₄), a major greenhouse gas, from submerged paddy soils is generally reduced by introducing intermittent drainage in summer, which is a common water management in Japan. However, such a practice is not widely conducted in Hokkaido, a northern region in Japan, to prevent a possible reduction in rice grain yield caused by cold weather. Therefore, the effects of intermittent drainage on CH₄ emission and rice grain yield have not been investigated comprehensively in Hokkaido. In this study, we conducted a three-year field experiment in Hokkaido and measured CH₄ and nitrous oxide (N₂O) fluxes and rice grain yield to elucidate whether the reduction in CH₄ emission can be achieved in Hokkaido as well as other regions in Japan. Four experimental treatments, namely, two soil types [soils of light clay (LiC) and heavy clay (HC) textures] and two water management [continuous flood irrigation (CF), and intermittent drainage (ID)], were used, and CH₄ and N₂O fluxes were measured throughout the rice cultivation periods from 2016 to 2018. Cumulative CH₄ emissions in 2016 were markedly low, suggesting an initially low population of methanogens in the soils presumably due to no soil submergence or crop cultivation in the preceding years, which indicates a possible reduction in CH₄ emission by introducing paddy-upland crop rotation. Cumulative CH₄ emissions in the ID-LiC and ID-HC plots were 21–91% lower than those in the CF-LiC and CF-HC plots, respectively, whereas the cumulative N₂O emissions did not significantly differ between the different water managements. The amount of CH₄ emission reduction by the intermittent drainage was largest in 2018, with a comparatively long period of the first drainage for 12 days in summer. Rice grain yields did not significantly differ between the different water managements for the entire 3 years, although the percentage of well-formed rice grains was reduced by the intermittent drainage in 2018. These results suggest that CH₄ emission from paddy fields can be reduced with no decrease in rice grain yield by the intermittent drainage in Hokkaido. In particular, the first drainage for a long period in summer is expected to reduce CH₄ emission markedly.     

Differentiation-inducing effects of small fruit juices on HL-60 leukemic cells

Epidemiological studies indicate that high intakes of fruits and vegetables are associated with a reduced risk of cancer, and several plant-derived drugs have been developed in medical oncology. Since only a small part of the flora has been tested for any kind of bioactivity, we chose small fruits as sources of differentiation-inducing activity against HL-60 leukemic cells. We have prepared juices from various small fruits that grow mainly in the northern part of Japan. Screening of 43 samples indicated that juices of Actinidia polygama Maxim., Rosa rugosa Thunb., Vaccinium smallii A. Gray, and Sorbus sambucifolia Roem. strongly induced differentiation of HL-60 cells to monocyte/macrophage characteristics in a concentration-dependent manner as indicated by histochemical and biochemical examinations.     

Effects of light quality on conidiophore formation of the melon powdery mildew pathogen <Emphasis Type="Italic">Podosphaera xanthii</Emphasis>

The lengths of conidiophores in fungal colonies of the melon powdery mildew pathogen Podosphaera xanthii Pollacci KMP-6 N cultured under greenhouse (natural) conditions differed markedly from those cultured in a growth chamber. We hypothesized that light wavelength was responsible for the differences in conidiophore length. In this study, we examined the effects of light-emitting diode (LED) irradiation (purple, blue, green, orange, and red light) and white light on colony development and conidiophore formation in KMP-6 N using a stereomicroscope and a high-fidelity digital microscope. Colonies on leaves were flat under greenhouse conditions and under red LED light irradiation but were stacked under growth chamber conditions and under purple, blue, green, and orange LED light irradiation. In addition, KMP-6 N formed catenated conidia comprising six conidia per conidiophore under greenhouse conditions and red light but more than seven conidia per conidiophore under growth chamber conditions and purple, blue, green, and orange light. Furthermore, almost none of the conidia on top of the conidiophores grown under blue light were fully constricted. Therefore, these fungi could not scatter their conidia and spread infection. This is the first report of the effects of LED lights on conidiophore formation in the melon powdery mildew fungus P. xanthii. The results provide insight into the mechanisms underlying the responses of conidiophores to light of specific wavelengths and conidial scatter from conidiophores of melon powdery mildew fungi.     

Culture method and growth characteristics of marine benthic dinoflagellate Ostreopsis spp. isolated from Japanese coastal waters

Blooms of toxic dinoflagellates of the genus Ostreopsis, which is known as a producer of palytoxin (PTX) analogs, may pose a threat to human health in tropical, subtropical, and temperate regions around the world. In the present study, we established a suitable culture method for Ostreopsis spp. isolated from Japanese coastal waters and characterized their growth potential using the method to discuss their bloom dynamics. Each clonal strain of Ostreopsis cf. ovata, Ostreopsis sp.?1, Ostreopsis sp.?5, and Ostreopsis sp.?6 was incubated in 25?×?150?mm test tubes with a flat bottom containing various kinds of medium. Since Ostreopsis spp. strains grew well in IMK and/or f/2 media, we selected these media for cultivation of all the Ostreopsis spp. isolates. Growth rates of O. cf. ovata (0.834?divisions/day), Ostreopsis sp.?1 (0.619?divisions/day), and Ostreopsis sp.?6 (1.04?divisions/day) that produce PTX analogs significantly differed (p?<?0.05) respectively and are clearly higher than those of other reported epiphytic dinoflagellate Gambierdiscus toxicus, Prorocentrum lima, and Coolia monotis cultures, which suggest that these species have ecological advantages to predominate through the algal succession in Japanese coastal waters, resulting in a potential risk to human health in this region.     

Host range and properties of <Emphasis Type="Italic">Tomato chlorotic dwarf viroid</Emphasis>

We characterised the host range and physical properties of Tomato chlorotic dwarf viroid. Among the 46 plant species inoculated with the viroid, two in the family Compositae and 23 in the family Solanaceae were found to be systemic hosts. The viroids in the crude sap from diseased tomato plants were thermally inactivated by heating to 100°C for at least 40 min. These viroids also lost their infectivity when diluted in phosphate buffer to at least 10⁻⁶, or after 3 days of incubation at room temperature. However, the infectivity of the viroids in dried crude sap from the plants persisted throughout the 50-day test period.     

Cardiopulmonary and sedative effects of intravenous or epidural methadone in conscious dogs

Cardiopulmonary and sedative effects of intravenous or epidural methadone were compared. Six beagles were randomly assigned to group MIV (methadone 0.5 mg/kg IV + NaCl 0.9% epidurally) or MEP (methadone 0.5 mg/kg epidurally + NaCl 0.9% IV). Cardiopulmonary, blood gas and sedation were assessed at time (T) 0, 15, 30, 60, 120, 240 and 480 min after drug administration. Compared to T0, heart rate decreased at T15–T120 in MIV (p < .001) and T15–T240 in MEP (p < .05); mean arterial pressure was reduced at T15–T60 in MEP (p < .01); respiratory rate was higher at T15 and T30 in both groups (p < .05); pH was lower at T15–T120 in MIV (p < .01) and T15, T30 and T120 in MEP (p < .05); PaCO₂ was higher at T15–T60 in MIV (p < .01) and T15, T30 and T120 in MEP (p < .01); sedation scores were higher at T15 and T30 in MIV and T15–T60 in MEP (p < .05). At T120 and T240, sedation score was higher in group MEP compared with group MIV (p < .01) In conclusion, cardiopulmonary and sedative effects of identical methadone doses are similar when administered IV or epidurally to conscious healthy dogs.     

Structure-Activity Relationship Study of the Neuritogenic Potential of the Glycan of Starfish Ganglioside LLG-3

LLG-3 is a ganglioside isolated from the starfish Linchia laevigata. To clarify the structure-activity relationship of the glycan of LLG-3 toward rat pheochromocytoma PC12 cells in the presence of nerve growth factor, a series of mono- to tetrasaccharide glycan derivatives were chemically synthesized and evaluated in vitro. The methyl group at C8 of the terminal sialic acid residue was crucial for neuritogenic activity, and the terminal trisaccharide moiety was the minimum active motif. Furthermore, the trisaccharide also stimulated neuritogenesis in human neuroblastoma SH-SY5Y cells via mitogen-activated protein kinase (MAPK) signaling. Phosphorylation of extracellular signal-regulated kinase (ERK) 1/2 was rapidly induced by adding 1 or 10 nM of the trisaccharide. The ratio of phosphorylated ERK to ERK reached a maximum 5 min after stimulation, and then decreased gradually. However, the trisaccharide did not induce significant Akt phosphorylation. These effects were abolished by pretreatment with the MAPK inhibitor U0126, which inhibits enzymes MEK1 and MEK2. In addition, U0126 inhibited the phosphorylation of ERK 1/2 in response to the trisaccharide dose-dependently. Therefore, we concluded that the trisaccharide promotes neurite extension in SH-SY5Y cells via MAPK/ERK signaling, not Akt signaling.