Suppression of conidial germination and appressorial formation by silicate treatment in powdery mildew of strawberry

The mode of action of soluble silicon against strawberry powdery mildew (Sphaerotheca aphanis var. aphanis) was investigated in four experiments. First, silicon-treated leaves from plants grown with silicate (Si+) and control leaves were excised, inoculated with conidia, and subsequent germination and formation of appressoria in a petri dish was assessed after 24 h. The germination rate was 49.7% on Si+ leaves, and was 67.2% on control leaves (t-test, P < 0.01). Second, we soaked cellulose membranes in various solvents and then placed the membranes on 4% water agar, dusted the membranes with conidia, and examined after 12 h. No difference was apparent between any treatment and the control (distilled water). Third, strawberries growing hydroponically with additional silicon in the medium were inoculated with conidia, and leaves were observed with a scanning electron microscope 1–2 days after inoculation. Germ tubes and secondary hyphae were shorter and had fewer branches on Si+ leaves than on the control. Moreover, penetration appeared to be inhibited. Fourth, the cuticle was separated from leaves from plants grown as in the third experiment, placed on water agar, and dusted with conidia. Germination of conidia, observed with a light microscope, on Si+ leaves was suppressed markedly to 40%–60% of that of the control. These results suggested that soluble silicon induced physiological changes in the cuticle layer after absorption by the plant. In addition, soluble silicate reduced germination of conidia, formation of appressoria, and possibly the penetration of powdery mildew.     

A new <Emphasis Type="Italic">Rhizoctonia</Emphasis> sp. closely related to <Emphasis Type="Italic">Waitea circinata</Emphasis> causes a new disease of creeping bentgrass

Isolates of an unidentified Rhizoctonia sp. (UR isolates) were obtained from creeping bentgrass and Kentucky bluegrass with reddish brown sheath and foliar rots. Because the UR isolates anastomosed with isolates of three varieties of Waitea circinata (var. oryzae, var. zeae, and var. circinata), colony morphology, hyphal growth rate at different temperatures, pathogenicity, sequence analysis of the internal transcribed spacers (ITS) region of ribosomal RNA genes (rDNA) were compared. The colony color of mature UR isolates was distinct from isolates of the other three varieties of W. circinata. In pathogenicity tests on creeping bentgrass, the severity of the disease caused by UR isolates was significantly higher than that caused by the three varieties of W. circinata. Sequence similarities of the rDNA-ITS region between UR isolates and between isolates within each variety were high (97–100%), but they were lower among isolates from UR and the varieties of W. circinata (88–94%). In a phylogenetic tree based on the rDNA-ITS sequences, UR isolates formed a cluster separate from each of the clusters formed by the three varieties of W. circinata. These results indicate that the UR isolates clearly differ from the three varieties of W. circinata. We therefore propose that the UR isolates be classified as new Rhizoctonia sp. that are closely related to W. circinata and that the disease on creeping bentgrass should be called Waitea reddish-brown patch disease (Sekikasshoku-hagusare-byo in Japanese).     

Role of the α subunit of heterotrimeric G-protein in probenazole-inducing defense signaling in rice

 Certain cellular responses to stresses and stimuli are regulated by a G-protein-mediated signaling pathway. A rice dwarf mutant that is defective in the α subunit of the heterotrimeric G-protein was found to be fully protected from blast fungus by the plant activator probenazole (PBZ) despite the 1-day delay in induction of the PR-10 gene PBZ1 by PBZ. These results suggest that the signaling pathway for protection by PBZ is not via the G-protein, although G-protein is involved in the induction of PBZ1 by PBZ. Received: March 27, 2002 / Accepted: August 20, 2002     

An avian pathogenic Escherichia coli strain produces a hemolysin, the expression of which is dependent on cyclic AMP receptor protein gene function

An avian pathogenic Escherichia coli strain M1000 showed a clear zone of erythrocyte lysis on sheep blood agar plates. The hemolytic activity was not detected in the culture supernatant nor was any DNA sequence homologous to the E. coli -hemolysin gene detected in the chromosome or plasmid DNA of the strain, indicating that the observed hemolysis was different from -type. To identify the genetic determinant responsible for the hemolysis, we performed random Tn5 insertional mutagenesis and obtained one mutant, named M5005, that totally lacked the hemolytic activity. Cloning and nucleotide sequencing of the region flanking the transposon insertion site in the M5005 chromosome revealed that the transposon was inserted within an open reading frame of the cyclic AMP receptor protein (CRP) gene, which is involved in one of the global regulatory networks of gene expression in E. coli. Nucleotide sequence analysis of the intact crp gene of the strain M1000 showed that the CRP protein of M1000 is 99% identical to that of K-12. Introduction of the intact crp gene on a low copy plasmid into the mutant M5005 restored the hemolytic phenotype, confirming that the mutation site in M5005 was in the crp gene. CRP plays a central role in catabolite repression, the phenomenon by which the synthesis of many enzymes required to metabolize various sugars is repressed in the presence of glucose. When the hemolytic activity of E. coli M1000 grown in the presence of glucose was examined, the hemolysis was totally impaired. These results indicate that the avian pathogenic E. coli strain M1000 produces a hemolysin the expression of which is dependent on crp gene function.     

Effects of pre-slaughter nutritional condition on intramuscular collagen solubility, pyridinoline cross-links and meat tenderness in aged goats

Effects of preslaughter nutritional condition on intramuscular collagen characteristics were studied, in order to clarify the potential of intensive feeding to aged animals in improving meat tenderness. Ten castrated male goats were assigned into one of two groups: one group was allowed ad libitum access to a concentrate diet (total digestible nutrients 70%, crude protein 15%) and Italian ryegrass hay (ADLIB‐group), and the other group was fed a restricted amount of their diet (concentrate diet 0.5% of bodyweight/day; hay 1.5% of bodyweight/day) to maintain their bodyweight (MAIN‐group). After 3 months of the experimental period, goats were slaughtered and meat samples were obtained immediately. Goats in ADLIB‐group had lower total and insoluble collagen concentrations, higher fat concentrations and collagen solubility than those in MAIN‐group, but soluble collagen concentrations of muscles were similar for both groups. Goats in ADLIB‐group had lower pyridinoline concentrations than those in MAIN‐group, in all muscles, but the differences of pyridinoline concentration between the groups were not statistically significant. Warner‐Bratzler shear force values of Longissimus and Biceps femoris muscles were lower in ADLIB‐group than in MAIN‐group. The increase of meat tenderness by preslaughter intensive feeding seemed to be associated with the increase in intramuscular fat deposition and high collagen solubility, and pyridinoline cross‐link appeared to be one of the factors related to collagen solubility.     

Development of immune assay system for both CpG and non-CpG DNA from lactic acid bacteria using a transfectant of swine Toll-like receptor 9

Bacterial DNA is expected to be a potent immune stimulating agent to Toll‐like receptor (TLR) 9 expressed cells such as macrophages, monocytes, B lymphocytes, NK cells and dendritic cells. In the present study, we constructed a transfectant of swine TLR9 with mammalian cells. We demonstrated that the transfectant, recognizing both CpG and non‐CpG oligonucleotides from lactic acid bacteria, induced NF‐κB activation by gene reporter assay. These findings indicate that the swine TLR9 transfectant will be highly useful for the screening of immunostimulatory DNA from lactic acid bacteria.     

Proteome analysis of cashmere

As an analysis of the cashmere proteins by Type IV 2‐DE, ten kinds of components, including three components with molecular mass 42–50 kDa whose expression level increased in the winter, were separated. In analyzing nine components of these ten using a mass spectrometer, the three components of molecular mass 70–120 kDa and pI 5.3 were identified as keratin type II microfibrillar (accession no. KRSHL2 ), keratin 48 k type I microfibrillar component 8c‐1 (accession no. KRSHL1 ) and cytosolic phospholipase A2 (accession no. O77793 ), respectively. The three components whose expression level increased in the winter, were identified as keratin type I microfibrillar 48 kDa component 8C‐1 (accession no. P02534 ) and keratin type I microfibrillar 47.6 kDa (accession no. P25690 ) (pI 5.2/42 kDa), keratin type II microfibrillar component 7C (accession no. P15241 ) and keratin typeII‐sheep (accession no. S34165 ) (pI 5.5/45 kDa), and the keratin type II microfibrillar component 5 (accession no. P25691 ) (pI 5.8–6.0/45 kDa), respectively. The three components of less than 17 kDa were identified as hair keratin type II intermediate filament (accession no. CAA51836 ) (pI 5.2) and keratin high‐sulfur matrix protein IIIB2 (accession no. P02447 ) with a different isoelectric point (pI 5.4 and 5.9), respectively.     

Characterization of new photosynthesis-inhibiting imidazolidine derivatives. 2. Interpretation of pre-emergence herbicidal performance from photosyghesis-inhibiting activity and soil adsorption coefficient

Ruling factors governing pre-emergence herbicidal activity were analysed for 16 photosynthesis-inhibiting 5-hydroxy-3-methyl-2–oxo-imidazolidine-1-carboxamide derivatives. Herbicidal performance was quantified by the reduction in area of experimental weed vegetation, measured by a computer-aided image analysis system. A system for fluorometric estimation of photosynthesis inhibitor concentration in aqueous solution greatly facilitated determination of the soil adsorption coefficients (K_d). Maximum herbicidal performance was found for N-sec-butyl-5-hydroxy-3-methyl-2-oxo-imidazolidine-1-carboxamide, a compound with the second lowest soil adsorptivity and average photosynthesis-inhibiting activity. A multiple regression analysis suggested that herbicidal performance of the soil-applied imidazolidine derivatives was determined by a balance between K_d and photosynthesis-inhibiting activity. In the present experimental system, however, the main influence was attributed to K_d.     

Lipophilic,hydrolytic and photolytic properties and fungicidal activity against Botrytis cinerea of 1-(4-substituted phenoxymethyl)-2, 2-dimethylpropyl imidazole-1-carboxylates and -thiocarboxylates

The sterol 14α-demethylation inhibitors 1-(4-substituted phenoxymethyl)-2, 2-dimethylpropyl imidazole-1-carboxylates and their corresponding thiocarboxylates were optimized for maximum in-vitro activity against Botrytis cinerea Pers. ex Fr. in terms of the lipophilic parameter π. The activity of both carboxylates and thiocarboxylates was strongly related with π and predicted to be maximum at π 1.38 and 0.72 respectively. However, the preventive efficacy of the carboxylates against B. cinerea on Solanum melongena L. grown in a greenhouse did not correlate with the parameter. Despite the unfavourable lower lipophilicity, compounds with p-alkoxy substituents were superior to those with any other substituent. Moreover, the p-methoxy-substituted thiocarboxylate 46 was much less effective in greenhouse tests than the corresponding carboxylate 15, despite their equivalent in-vitro activity. To clarify these discrepancies, hydrolytic and photolytic stabilities of several representative compounds including 15 and 46 were investigated. The carboxylates examined were much more labile to hydrolysis than the thiocarboxylate 46; however, the four thiocarboxylates studied were less stable to light than the carboxylates. Consequently, the above discrepancies were attributable mainly to the superior stability of the carboxylates to photolysis as compared to the thiocarboxylates.