Overexpression of a fungal laccase gene induces nondehiscent anthers and morphological changes in flowers of transgenic tobacco

Laccases play important roles in the development of fruiting bodies and in lignin degradation by basidiomycetes. In this study, we present novel phenotypes of transgenic tobacco plants with a chimeric gene for fungal laccase under the control of the cauliflower mosaic virus 35S promoter. At the flowering stage, the transgenic plants that produced recombinant laccase had brownish anthers instead of the greenish anthers of wild-type plants. The brownish anthers exhibited male sterility with a nondehiscent phenotype at varying frequencies. The frequency of nondehiscence depended on the temperature at which plants were cultivated and it was higher at 24°C than at 29°C. The cell wall structures of transgenic anther tissues were almost the same as in the wild type, but the stomium was severely deformed, and abnormal components were apparent in cells of the endothecium and epidermis. Furthermore, the pattern of deposition of flavonoids in the transgenic anther epidermis differed from the wild-type pattern. The expression of laccase also induced other phenotypic changes in the flowers of transgenic plants, namely, increased petal number, fused and petaloid stamens, and doubling of floral organs. These results indicate that the ectopic expression of laccase influences various aspects of flower development.     

Root pruning is effective in alleviating the inhibition of soybean growth caused by anaerobic stress for a short period

Soybean is an important upland crop,but its productivity is often limited by anaerobic stress caused by waterlogging.The ability to adjust root growth under environmental constraints is an important physiological trait for adapting to an everchanging environment,and root pruning is an artificial technique for regenerating the root system.In the present study,we investigated whether root pruning in soybean can effectively alleviate the inhibitory effects of anaerobic stress.Soybean plants were af...     

Identification of overwintering vegetative cells of the bivalve-killing dinoflagellate <Emphasis Type="Italic">Heterocapsa circularisquama</Emphasis> in Uranouchi Inlet,Kochi Prefecture,Japan

ABSTRACT:   Red tides of Heterocapsa circularisquama have led to serious damage of bivalve aquacultures in western coastal areas of Japan. To understand the whole picture regarding the ecology of this species, it is essential to clarify its overwintering mechanisms. In this study, the population dynamics of H. circularisquama were investigated from February 2004 to November 2005, and overwintering cells were identified for the first time in water columns of Uranouchi Inlet, Kochi Prefecture, Japan. Heterocapsa circularisquama cells were detected by the indirect fluorescent antibody technique using monoclonal antibodies that specifically recognize and react to this species. Vegetative cells were almost always detected from the first observation in February 2004 to November 2005 with temperatures of 10.5–30.6°C. During the period from winter to spring, this species survived in areas with a temperature higher than 10°C. The overwintering cells of H. circularisquama were isolated in March 2004, and identification was made via observation of the morphology and body scales of the cultured cells. These overwintering cells were identified as H. circularisquama and reacted to the monoclonal antibody. These results indicate that H. circularisquama can overwinter and survive throughout the year in a vegetative cell state in Uranouchi Inlet.     

Effects of cultivation of preceding plants on the development of Fusarium wilt of cucumber and daikon

The effects of the cultivation of preceding plants on the inhibition of Fusarium wilts, vessel diseases of cucumber (Cucumis sativus) caused by Fusarium oxysporum f. sp. cucumerinum (FOC) and yellows of daikon (Raphanus sativus var. longipinnatus) caused by F. oxysporum f. sp. raphani (FOR), were examined. When the cucumber or daikon plant was cultivated as a succeeding plant in the pots where the soils had been inoculated with a spore suspension of FOe or FOR and then various preceding plants had been cultivated, the percentage of disease of the succeeding plants was lower in some plots than in the control plot without preceding plants. Spinach (Spinacia oleracea, cv. Atlas) and burdock (Arctium lappa, cv. Takinogawa) inhibited significantly the vessel disease caused by FOe. Thrnip (Brassica rapa, Parental Line Nou No.1) and spinach (S. oleracea, cv. Try) suppressed the yellows caused by FOR. When the number of FOC or FOR was estimated by the recovery using Komada med.ium and by testing their pathogenicity to the host plant, cucumber or daikon, the numbers of both were not reduced in the plots even where these plants had been cultivated initially. The soil extract from the pots, where preceding plants had been cultivated, did not affect the conidia of FOC and FOR. In both FOC and FOR experiments, the cultivation of preceding plants increased the number of bacteria in soil. Additionally, in the FOR experiment, preceding plants, turnip and spinach, increased the number of F. oxysporum (FOX) except for FOR and decreased the proportion of FOR to FOX. It was suggested that these preceding plants might affect the pathogens of Fusarium wilt indirectly by the changes in the numbers and flora of soil microorganisms.     

Recovery of leptospires from miniature pigs experimentally infected with Leptospira interrogans serovar Manilae strain UP-MMC under immunosuppressive conditions by dexamethasone

Leptospira interrogans serovar Manilae strain UP-MMC was inoculated into miniature pigs to assess its pathogenicity. Leptospires were recovered from the whole blood, kidneys, and livers in the acute phase without showing any clinical signs. Under immunosuppressive conditions by dexamethasone, leptospires were recovered from the kidneys and their genes were detected from the urine in the chronic phase. These results indicate that leptospires persisted in the kidneys until the chronic phase, and excretion of leptospires in the urine was enhanced under immunosuppressive conditions, resulting in horizontal transmission among pigs on farms.     

Enhancement of Drug Resistance by Lysophosphatidic Acid Receptor-3 in Mouse Mammary Tumor FM3A Cells

Lysophosphatidic acid (LPA) acts as a simple phospholipid that interacts with G protein-coupled transmembrane LPA receptors. Recently, it has been reported that each LPA receptor plays different biological roles in acquisition of the malignant property of tumor cells. In this study, to assess the involvement of LPA receptor-3 (LPA₃) in cell survival after treatment with anticancer drugs, we generated Lpar3-expressing FM3A-a3A9 cells from mouse mammary tumor FM3A cells and examined the cell survival rate after treatment with anticancer drugs compared with Lpar3-unexpressing cells. Cells were treated with 0.005 to 10 μM of cisplatin (CDDP) or doxorubicin (DOX) for 3 days. For the CDDP and DOX treatments, the cell survival rate of FM3A-a3A9 cells was significantly higher than that of Lpar3-unexpressing cells. The expression level of the Mdr1a gene in FM3A-a3A9 cells was higher than that of Lpar3-unexpressing cells, whereas no significant difference in multidrug resistance 1b (Mdr1b) and glutathione S-transferase mu1 (Gstm1) expressions was found. These results suggest that LPA₃ may enhance the cell survival rate after treatment with anticancer drugs in mouse mammary tumor cells, correlating with increased expression of the Mdr1 gene.     

Food-grade mulberry powder enriched with 1-deoxynojirimycin suppresses the elevation of postprandial blood glucose in humans

Mulberry 1-deoxynojirimycin (DNJ), a potent glucosidase inhibitor, has been hypothesized to be beneficial for the suppression of abnormally high blood glucose levels and thereby prevention of diabetes mellitus. However, DNJ contents in commercial mulberry products were as low as about 0.1% (100 mg/100 g of dry product), implying that the bioavailability of DNJ might not be expected. We carried out studies in two directions: (1) production of food-grade mulberry powder containing a maximally high DNJ content; (2) determination of the optimal dose of the DNJ-enriched powder for the suppression of the postprandial blood glucose through clinical trials. The following method was used: (1) DNJ concentrations in mulberry leaves from different cultivars, harvest seasons, and leaf locations were determined using hydrophilic interaction chromatography with evaporative light scattering detection. (2) Healthy volunteers received 0, 0.4, 0.8, and 1.2 g of DNJ-enriched powder (corresponding to 0, 6, 12, and 18 mg of DNJ, respectively), followed by 50 g of sucrose. Before and 30-180 min after the DNJ/sucrose administration, plasma glucose and insulin were determined. The following results were obtained: (1) Young mulberry leaves taken from the top part of the branches in summer contained the highest amount of DNJ. After optimization of the harvesting and drying processes for young mulberry leaves (Morus alba L. var. Shin ichinose), DNJ-enriched powder (1.5%) was produced. (2) A human study indicated that the single oral administration of 0.8 and 1.2 g of DNJ-enriched powder significantly suppressed the elevation of postprandial blood glucose and secretion of insulin, revealing the physiological impact of mulberry DNJ (effective dose and efficacy in humans). This study suggests that the newly developed DNJ-enriched powder can be used as a dietary supplement for preventing diabetes mellitus.     

Immuno-magnetic separation and agar layer methods for the isolation of freeze-injured Yersinia enterocolitica O:8 from water

To develop an effective method to isolate an injured pathogenic Yersinia enterocolitica O:8 organism from environmental samples, we compared the isolation of freeze-injured and non-injured Y. enterocolitica O:8 and found that the isolation was more successful when immuno-magnetic separation (IMS) with anti-Y. enterocolitica O:8 antibody was used. Plating onto cefsulodin-irgasan-novobiocin (CIN) agar and Virulent Yersinia enterocolitica (VYE) agar by means of the agar layer method was found to be effective in isolating the injured cells. The alkali treatment which is generally used for selective detection of Yersinia organism failed to isolate freeze-injured pathogenic Y. enterocolitica O:8 cells. Recovery methods without using the alkali treatment were superior for detecting freeze-injured Y. enterocolitica O:8. Our results demonstrate that the IMS and the agar layer methods should be used to isolate injured pathogenic Yersinia organisms from environmental samples such as water.     

Nonsteroidal anti-inflammatory drugs induce hypermotilinemia and disturbance of interdigestive migrating contractions in instrumented dogs

Nonsteroidal anti-inflammatory drugs (NSAIDs) induce gastric ulcers due to inhibition of prostaglandin production. Prostaglandins have an influence on physiological gastrointestinal motility, but the relationships between NSAID-induced gastric ulcer, gastrointestinal motility and motilin are unknown. Fifteen dogs were allocated randomly to three groups in which either gelatin, meloxicam or indomethacin was administered. Fecal occult blood and gastrointestinal motility were monitored continuously for 6 days. In addition, analyses of the plasma motilin concentration, gastrointestinal endoscopy and gastric emptying, and detection of motilin cells were performed. Gastrointestinal motility was disturbed in the indomethacin group, presenting as disappearance of interdigestive migrating contractions (IMCs) 3 days before gastric ulcers were detected. Delayed gastric emptying and hypermotilinemia were observed significantly more often in the indomethacin group compared with the other groups. Motilin cell-crypt/villi ratio in the indomethacin group significantly decreased in the duodenum and jejunum, compared with the other groups. No significant changes in any tests were observed in the meloxicam group, when compared with the gelatin group. These findings suggest that the disturbance of IMCs caused by hypermotilinemia, with changes in motilin cell distribution, and delayed gastric emptying induced by indomethacin may contribute to the development of gastric ulcers.