Histopathology of red crown rot of soybean

Histopathological analyses of red crown rot (RCR) of soybean were performed using field-grown and indoor-grown soybean plants. These analyses revealed that soybean plants formed a periderm to prevent pathogen invasion. When a break formed in the periderm, an additional periderm newly differentiated into the inner cells and prevented pathogen invasion. If the pathogen reached the intravascular tissues, some xylem parenchyma cells died, and the vessels were occluded by gum. Periderm formation and vessel occlusion were observed not only in slightly diseased soybean plants, but also in seriously diseased plants, indicating that both reactions function as part of the defense response against RCR.     

Serum from dogs Infected with Babesia gibsoni inhibits maturation of reticulocytes and erythrocyte 5'-nucleotidase activity in vitro

Erythrocyte 5'-nucleotidase is thought to be involved in the maturation of erythrocytes. In the present study, in vitro incubation of canine erythrocytes demonstrated that significant inhibition of 5'-nucleotidase activity occurred in the presence of serum from dogs infected with Babesia gibsoni, when the enzyme was assayed with cytidine 5'-monophosphate (5'-CMP) and inosine 5'-monophosphate (5'-IMP) as substrates. The multiplication of B. gibsoni in in vitro culture also resulted in a significant decrease in the enzyme activity of erythrocytes in the culture. Furthermore, the infected serum and 5'-CMP retarded the maturation of canine reticulocytes in vitro. These results suggested that nucleotides such as 5'-CMP and 5'-IMP might accumulate in young erythrocytes and/or serum in dogs infected with B. gibsoni as a result of decreased activity of erythrocyte 5'-nucleotidase, resulting in the delayed maturation of reticulocytes.     

<Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>-mediated transformation for random insertional mutagenesis in <Emphasis Type="Italic">Colletotrichum lagenarium</Emphasis>

Random insertional mutagenesis using a marker DNA fragment is an effective method for identifying fungal genes relevant to morphogenesis, metabolism, and so on. Agrobacterium tumefaciens-mediated transformation (AtMT) has long been used as a tool for the genetic modification of a wide range of plant species. Recent study has indicated that A. tumefaciens could transfer T-DNA not only to plant cells but also to fungal cells. In this study, AtMT was applied to Colletotrichum lagenarium for random insertional mutagenesis. We constructed a binary vector pBIG2RHPH2 carrying a hygromycin-resistant gene cassette between the right and left borders of T-DNA. Optimal co-cultivation of C. lagenarium wild-type 104-T with pBIG2RHPH2-introduced A. tumefaciens C58C1 led to the production of 150–300 hygromycin-resistant transformants per 10⁶ conidia. Southern blot analysis revealed that T-DNA was mainly integrated at a single site in the genome and at different sites in transformants. The T-DNA inserts showed small truncations of either end, but the hygromycin-resistant gene cassette inside the T-DNA was generally intact. The mode of T-DNA insertion described above resulted in highly efficient gene recovery from the transformants by thermal asymmetrical interlaced-polymerase chain reaction. The fungal genomic DNA segments flanking T-DNA were identified from five of eight mutants that had defective melanin biosynthesis. The sequence from one of the segments was identical to that of the melanin biosynthesis gene PKS1 of C. lagenarium, which we previously characterized. These results strongly support our notion that AtMT is a possible tool for tagging genes relevant to pathogenicity in the plant pathogenic fungus C. lagenarium.     

Evaluation of the Endophyte Enterobacter cloacae SM10 Isolated from Spinach Roots for Biological Control against Fusarium Wilt of Spinach

Six hundred sixty-three isolates of microorganisms, including fungi and bacteria, were collected from surface-sterilized roots of spinach (Spinacia oleracea L.) growing in commercial greenhouses in Kyoto Prefecture. These isolates were screened for their ability to control Fusarium wilt of spinach caused by Fusarium oxysporum f. sp. spinaciae. In primary screening, spinach seeds were treated with the isolates, sown in pots containing sterilized soil, and then challenge-inoculated with the pathogen. Nine bacteria were effective in reducing disease incidence. Subsequently, spinach seeds were treated with the selected isolates, then sown in an infested field and grown from June to July 1998. Four bacteria reduced disease incidence. One of these four, designated as SM10, significantly suppressed the disease. Based on bacteriological properties, SM10 was identified as a strain of Enterobacter cloacae. SM10 was observed within xylem vessels of spinach roots using light and immunoelectron microscopy, indicating E. cloacae SM10 was an endophytic bacterium of spinach. Received 4 July 2000/ Accepted in revised form 13 September 2000     

Protective effect of a lignan-containing flaxseed extract against CCl(4)-induced hepatic injury

Carbon tetrachloride (CCl(4)) -induced hepatotoxicity is a commonly used model for investigating lipid peroxidation-related tissue injury. In the present study, the effect of flaxseed extract was observed on histological sections, glutathione-content and DNA strand breaks. Lignan-containing flaxseed extract (1.6 g/kg body weight/day) was daily administered with intragastric injection to rats for three days, on the fourth day, CCl(4) (2 g/kg) was intraperitoneally injected. Liver tissue was sampled at 24 hr after administering CCl(4). Liver-necrosis was observed in CCl(4)-injected rats without pretreatment of flaxseed extract. Pretreatment of flaxseed extract reduced extent of the necrosis found 24 hr after the intraperitoneal administration of CCl(4). Pretreatment of flaxseed extract protect against CCl(4)-induced decrease of reduced glutathione-content measured from reactions with 5,5'-dithiobis-(2-nitrobenzoic acid) and also protect against the elevation of DNA strand breaks in the liver cells measured by comet assay. Flaxseed-extract appears to protect liver cells against CCl(4)-induced necrosis.     

Characterization of Grapevine Algerian latent virus isolated from nipplefruit (Solanum mammosum) in Japan

Alfalfa mosaic virus (AMV), Cucumber mosaic virus (CMV), Potato virus Y (PVY), Tomato bushy stunt virus nipplefruit strain (TBSV-Nf), and an unknown spherical virus were isolated from nipplefruit (Solanum mammosum) cultivated in Chiba Prefecture, Japan. The spherical virus was identified as Grapevine Algerian latent virus nipplefruit strain (GALV-Nf) from the genus Tombusvirus, based on its physical properties, serological relationships, and analysis of genomic RNA. The genomic RNA of GALV-Nf is 4731 nucleotides long and encodes five open reading frames as well as those of other tombusviruses. Nipplefruit infected with GALV-Nf had severe stunting, leaf deformation, and clear mosaic symptoms. This is the first report of an isolation of GALV in Japan. An erratum to this article is available at .     

Incidence of viruses in <Emphasis Type="Italic">Alstroemeria</Emphasis> plants cultivated in Japan and characterization of <Emphasis Type="Italic">Broad bean wilt virus-2, Cucumber mosaic virus</Emphasis> and <Emphasis Type="Italic">Youcai mosaic virus</Emphasis>

Alstroemeria plants were surveyed for viruses in Japan from 2002 to 2004. Seventy-two Alstroemeria plants were collected from Aichi, Nagano, and Hokkaido prefectures and 54.2% were infected with some species of virus. The predominant virus was Alstroemeria mosaic virus, followed by Tomato spotted wilt virus, Youcai mosaic virus (YoMV), Cucumber mosaic virus (CMV), Alstroemeria virus X and Broad bean wilt virus-2 (BBWV-2). On the basis of nucleotide sequence of the coat protein genes, all four CMV isolates belong to subgroup IA. CMV isolates induced mosaic and/or necrosis on Alstroemeria. YoMV and BBWV-2 were newly identified by traits such as host range, particle morphology, and nucleotide sequence as viruses infecting Alstroemeria. A BBWV-2 isolate also induced mosaic symptoms on Alstroemeria seedlings.     

Adsorption of OP<Subscript>1</Subscript>-related bacteriophages requires the gene encoding a TonB-dependent receptor-like protein in <Emphasis Type="Italic">Xanthomonas</Emphasis><Emphasis Type="Italic">oryzae</Emphasis> pv. <Emphasis Type="Italic">oryzae</Emphasis>

Xanthomonas oryzae pv. oryzae strain T7174R is lysed by bacteriophage OP_1h and OP_1h2. Three mutants tolerant to both OP_1h and OP_1h2 were isolated by transposon mutagenesis. The mutants had an insertion of the transposon in XOO1687, which is predicted to encode a TonB-dependent receptor gene. Plasmid pHMIroNB that contained XOO1687 of T7174R was constructed, and the mutant was transformed with the plasmid. The transformant recovered sensitivity to OP_1h and OP_1h2. Electron microscopic analysis demonstrated that OP_1h and OP_1h2 can adsorb to the wild type and the transformant, but they could not adsorb to the phage-tolerant mutant. These results suggest that the TonB-dependent receptor gene relates to adsorption and infection of T7174R by OP_1h2 and OP_1h. Y. Inoue and S. Tsuge have contributed equally to this work.     

Chemical components of hardwood barks stripped by the alien squirrel Callosciurus erythraeus in Japan

The Pallas squirrel (Callosciurus erythraeus; common name in Japan: Formosan squirrel) was introduced in the 1950s and has established populations throughout southeastern Kanagawa Prefecture, Japan. Some tree species in natural forests, parks, and gardens in residential areas have been intensively debarked by the squirrels, especially in winter and spring. The amounts of chemical components, such as polyphenols, resins, flavanols, and sugars, in the bark were compared among species and individual trees collected in the forest of Yokohama Nature Sanctuary. Interspecific differences in the extent of stripping were not related to the amounts of the four chemical components. For individual trees, the bark with feeding scars tended to contain more sugar than that without scars.