AAL-toxin,a potent natural herbicide which disrupts sphingolipid metabolism of plants

AAL-toxin, a product of Alternaria alternata (Fr.) Keissl., is effective as a herbicide at low concentrations against a range of broadleaf plants (e.g. jimsonweed, prickly sida and black nightshade). However, monocotyledonous crops such as maize and wheat, as well as some varieties of tomato, are tolerant to it. The IC50 values for cellular electrolyte leakage and chlorophyll loss in duckweed (Lemna pausicostata L.) after 72 h treatment were 20–40 nM. Similar results were obtained with a susceptible tomato variety. AAL-toxin caused rapid cellular leakage of electrolytes, followed by cellular collapse; the first symptom at the ultrastructural level is disruption of the plasma membrane. The effects of the toxin are not light-dependent and appear to be associated with dysfunction of the plasma membrane. Fumonisins and sphingoid bases such as phytosphingosine cause similar effects, although these compounds are less potent (fumonisins, about 10-fold; sphingoid bases, about 100-fold). Recent studies suggest that in duckweed and in susceptible tomato varieties, AAL-toxin- and fumonisin B1 -induced disruption of sphingolipid metabolism is an early event in the cascade of events leading to phytotoxic injury and cell death.     

Occurrence of strawberry marginal chlorosis caused by “Candidatus Phlomobacter fragariae” in Japan

In February 2004, a disease of strawberry (Fragaria × ananassa Duch.), causing little-leaf, proliferation, malformation of fruits, and marginal chlorosis of leaves, occurred in Ehime Prefecture, Japan. The causal pathogen was identified as a phloem-restricted bacterium-like organism, “Candidatus Phlomobacter fragariae,” based on polymerase chain reaction (PCR) detection, electron microscopy, and sequence analysis of PCR products. This is the first report of strawberry marginal chlorosis in Asia. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession number AB246669.     

Novel erythrocyte pits in the small tropical ruminant, lesser mouse deer

We examined unique erythrocyte pits of the peripheral blood and bone marrow in the lesser mouse deer, Tragulus javanicus, using scanning electron microscope (SEM) and transmission electron microscope (TEM). Under the SEM observation, the pit was observed as a hole on both mature erythrocytes of the peripheral blood and immature erythrocytes of the bone marrow. By the TEM, the mature erythrocytes had a vacuole, which showed complicated shape and occupied considerable space within the cytoplasm. The vacuole was communicated extracellularly by perforation, which corresponded to the hole on the cell surface. In the bone marrow, erythroblast and reticulocytes have a cytoplasmic vacuole. This abnormal feature of the erythrocytes is peculiar to the mouse deer, and not found in other tropical ruminants. Despite the disadvantage of volume loss from the small erythrocytes, the mouse deer were healthy and showed no signs of anaemia.     

Degradation and metabolism of a fungicide, 2,4,5,6-tetra-chloroisophthalonitrile (TPN) in soil

Summary Degradation of a fungicide, 2,4,5,6-tetrachloroisophthalonitrile (TPN) in soil was studied under laboratory conditions. TPN degraded more rapidly under 60% WHC conditions than at 20%, 40% and 100% WHC, while its degradation was rapid at temperatures of 25°C-30°C, evidently due to the microbial degradation. TPN degraded mainly through dechlorination and partly a substitution reaction. The degradation products identified by gas chromatographic analyses were: 2,4,5-trichloroisophthalonitrile (abbreviated as 2,4,5-Cl₃-IPN), 2,4,6-Cl₃-IPN, 2,4-Cl₂-lPN, 2,5-Cl₂-IPN, 4-Cl-IPN, 5-Cl-IPN, IPN, 2,5,6-Cl₃4-(OH)-IPN and 2,5,6-Cl₃-4-(OCH₃)-IPN. Peaks with longer retention times than that of TPN were not identified. Tentative degradation pathways were proposed on the basis of the identified degradation products. About 90% of the bacterial strains isolated from the soil to which TPN had been added degraded TPN, suggesting enrichment of the soil with TPN-degrading bacteria.Japanese International Cooperation Agency     

High temperatures activate local viral multiplication and cell-to-cell movement of Melon necrotic spot virus but restrict expression of systemic symptoms

The infection of melon plants by Melon necrotic spot virus (MNSV) and the development of necrotic disease symptoms are a seasonal occurrence in Japan, which take place between winter and early summer, but not during mid-summer. In this paper we investigate the effect of three different temperatures (15, 20, and 25 degrees C) on the local and systemic expression of MNSV in melon plants. Previously, the incidence of plants expressing systemic symptoms caused by MNSV and other viruses was found to be greater at temperatures less than 20 degrees C. In this study, our temperature-shift experiments support previous studies that found the expression of systemic symptoms increases as temperature falls from 25 to 20 degrees C and decreases as temperature rises from 20 to 25 degrees C. However, MNSV replication in melon cells and local viral movement within leaves following the inoculation of melon protoplasts or cotyledons were more frequent at 25 degrees C than at 15 or 20 degrees C.     

Development of cleaved amplified polymorphic sequence (CAPS)-based markers for identification of sweetpotato cultivars

To develop a simple and reliable method to identify sweetpotato cultivars, we designed cleaved amplified polymorphic sequence (CAPS)-based markers and used them to perform genotyping of Japanese sweetpotato cultivars. In order to screen the CAPS-based markers, 13 primer pairs were designed from the exon sequences of 11 sweetpotato genes to amplify fragments containing an intron. By digesting the amplified products with 8 restriction enzymes having different recognition sites, a total of 27 polymorphic marker fragments were obtained. Genotyping of 60 Japanese sweetpotato cultivars using these markers suggested that the markers can effectively distinguish sweetpotato cultivars. Among the genes used for primer design, the gene encoding the dihydroflavonol 4-reductase (DFR) showed the largest degree of polymorphism. To our knowledge, this is the first report on the development of CAPS-based markers in sweetpotato.     

Isolation of pathogenicity- and gregatin-deficient mutants of <Emphasis Type="Italic">Phialophora gregata</Emphasis> f. sp. <Emphasis Type="Italic">adzukicola</Emphasis> through <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>-mediated transformation

Phialophora gregata f. sp. adzukicola, a causal agent of brown stem rot in adzuki beans, produces phytotoxic compounds: gregatins A, B, C, D, and E. Gregatins A, C, and D cause wilting and vascular browning in adzuki beans, which resemble the disease symptoms. Thus, gregatins are considered to be involved in pathogenicity. However, molecular analyses have not been conducted, and little is known about other pathogenic factors. We sought to isolate nonpathogenic and gregatin-deficient mutants through Agrobacterium tumefaciens-mediated transformation (ATMT) for cloning of pathogenicity-related genes. The co-cultivation of P. gregata and A. tumefaciens for 48 h at 20°C with 200 μM acetosyringone resulted in approximately 80 transformants per 10⁶ conidia. The presence of acetosyringone in the A. tumefaciens pre-cultivation period led to an increase in T-DNA copy number per genome. Of 420 and 110 transformants tested for their pathogenicity and productivity of gregatins, one nonpathogenic and three gregatin-deficient mutants were obtained, respectively. The nonpathogenic mutant produced gregatins, whereas the gregatin-deficient mutants had pathogenicity comparable to the wild-type strain. This is the first report of ATMT of P. gregata. Further analysis of these mutants will help reveal the nature of the pathogenicity of this fungus including the role of gregatin in pathogenesis.