Identification and use of a wild plant with antimicrobial activity against Ralstonia solanacearum, the cause of bacterial wilt of potato

Fresh aerial tissue and roots of 14 wild plants in Okinawa prefecture were investigated for their antimicrobial activity against Ralstonia solanacearum , which causes bacterial wilt of potato. A 70% aqueous ethanol extract of fresh aerial tissue of Geranium carolinianum L. showed strong antimicrobial activity against R. solanacearum . This extract also showed antimicrobial activity against the pathogens causing common scab of potato and soil rot of sweet potato. The antimicrobial substance could be extracted with hot water, and was effective against R. solanacearum in soil. In the field test, a treatment combining incorporation of dried aerial tissue into the soil and solarization was highly effective for control of bacterial wilt of potato. These findings suggest that G. carolinianum L. could be used as a biological agent for the control of bacterial wilt of potato.     

TGF-β and TNF-α stimulate MMP-9 production in murine epidermal keratinocytes

Matrix metalloproteinases 2 and 9 (MMP‐2 and ‐9) are zinc‐dependent metalloenzymes and have gelatin‐degrading activity. Both MMP are known to be secreted by many types of cells and play important roles in several biological changes including tissue remodeling and wound healing. In the present study, a primary culture of murine epidermal keratinocytes was prepared and effects of transforming growth factor‐β (TGF‐β), tumor necrosis factor‐α (TNF‐α) and interferon‐γ (IFN‐γ) on expression of MMP‐2 and MMP‐9 by the keratinocytes was examined. Gelatin zymography revealed that murine epidermal keratinocytes secreted proenzyme forms of MMP‐2 and MMP‐9, but the active forms of both MMP were hardly detectable, indicating that in vitro autoactivation of these proenzymes did not occur. Both TGF‐β and TNF‐α stimulated MMP‐9 production in a dose‐dependent manner, but the MMP‐2 level was not changed. Interferon‐γ hardly affected production of MMP‐2 or MMP‐9. Ribonuclease protection assay demonstrated that TNF‐α increased the level of MMP‐9 mRNA 6‐fold compared to the control, whereas TGF‐β slightly up‐regulated it. These results suggest that expression of MMP‐9 could be regulated by several cytokines in murine epidermal keratinocytes.     

The abundance of juvenile yellowtail (Seriola quinqueradiata) near the Kuroshio: the roles of drifting seaweed and regional hydrography

We assess the effect of drifting seaweed (Sargassum sp.) biomass, geography and hydrography on juvenile yellowtail (Seriola quinqueradiata) abundance variation off the southeast coast of Japan, near the Kuroshio Current. The amount of drifting seaweed mats progressively increased northeastward into the cooler, coastal waters. Frontal structure indexed using a station‐to‐station ΔSST did not explain spatial variation in the seaweed mat distribution, although the western extent of the Kuroshio Current appeared to act as a boundary. Juvenile yellowtail constituted 51–62% of the fish collected in association with drifting seaweed mats in April 1996 and 1997 and 29% in June 1996. The abundance of juvenile yellowtail was positively correlated with seaweed biomass. The geographic distribution of juvenile yellowtail associated with drifting mats varied among sampling periods, being more southwesterly in April and more northeasterly in June. Simple multiple regression models based on seaweed biomass and geographic distribution (latitude) explained between 35% and 43% of the variation in juvenile yellowtail abundance in spring. Associations with spatial and temporal variations in hydrographic conditions did not contribute to explained variation in a meaningful manner. The results presented here indicate that, off the southeast coast of Japan during April, yellowtail juveniles are likely to be most abundant when seaweed biomass is high, occur offshore, and are bounded by the western extent of the Kuroshio Current near the 19–20°C SST isotherm.     

Expression of desmosomal proteins in rat keratinocytes during in vitro differentiation

The keratinocyte, the major component of the epidermis, expresses several proteins that characterize the keratinization during the differentiation. Proliferation and differentiation of cultured human keratinocytes are known to be regulated by the Ca2+ concentration in the culture medium. However, informations about the rat keratinocyte are relatively limited and their physiology is still an open question. To elucidate the characteristics of the rat keratinocyte, we established rat keratinocyte culture system and examined effects of extracellular calcium concentration on the expression of differentiation-related proteins. Keratinocytes were isolated from the newborn rat skin with 0.25% trypsin, followed by separation with a Percoll density gradient. The separated cells were grown in MCDB 153 medium containing several growth factors and Ca(2+)-free fetal bovine serum, then stimulated with Ca2+. Immunoblotting demonstrated strong expression of beta1 integrin in unstimulated cells, suggesting that the primary culture of rat keratinocytes was successfully established. Expression of desmoglein and transglutaminase was increased by Ca2+ stimulation, whereas beta1 integrin expression was decreased in response to increasing concentrations of Ca2+. These observations indicate that cultured rat keratinocytes maintain the ability to differentiate in vitro, which is similar to that of the basal keratinocytes in the epidermis.     

Spatial relationships among the cellular tapetum,visual streak and rod density in dogs

The dog visual system is well suited to dim light conditions due to rod-dominated retina and the reflective tapetum. The topographical distributions of rods and thickness of the tapetum of the dog were quantified in retinal whole mounts stained with thionine, and spatial relationships among the tapetum, rod density and visual streak of high ganglion cell density were elucidated. The relationship between the retina and tapetum was analyzed in parasagittal sections stained with thionine or hematoxylin-eosin. The tapetum was thick in its center, and the thickest part consisted of 9 to 12 tapetal cell layers. Rod density ranged from 200,000 to 540,000/mm². Maximum rod density was found in the area dorsal to the visual streak, and the density in that area was significantly higher than the rod density in the visual streak and accorded spatially with the thickest part of the tapetum. The horizontal visual streak was found over the horizontal line through the optic disc in the temporal half and extended slightly into the nasal half. The central area of the highest density of ganglion cells was approximately located midway between the nasal and temporal ends of the visual streak. The visual streak was located within the tapetal area, but ventrally to the thick part of the tapetum.     

Bacterial Blight of Welsh Onion : A New Disease Caused by Xanthomonas campestris pv. allii pv. nov.

In June of 1998, a new bacterial disease was observed on Welsh onion in Okinawa Prefecture, Japan. Infected plants in nursery boxes were stunted with tip dieback, and heavily infected plants died. In fields, the disease appeared on leaves as irregular gray spots or elliptical spots with creases in the center. These spots enlarged and spread rapidly continued cloudy or rainy weather, and formed blight lesions on outer leaves. Yellow mucoid bacterial colonies were consistently isolated from these lesions. The causal bacterium was identified as a pathovar of Xanthomonas campestris on the basis of bacteriological properties. The bacterium was pathogenic to Welsh onion, onion, but nonpathogenic to chive, Chinese chive and hyacinth. Of Liliaceae plants, which contain Welsh onion and onion, only hyacinth has been reported as a host for the genus Xanthomonas, namely X. campestris pv. hyacinthi. However, strains of X. campestris pv. hyacinthi were not pathogenic against either Welsh onion or onion. From these results, the bacterium isolated from Welsh onion is considered to be a new pathovar of X. campestris, and the name of X. campestris pv. allii pv. nov. is proposed. A strain MAFF 311173 is designated as the pathotype strain. Received 29 March 2000/ Accepted in revised form 4 July 2000