Flagellin from Pseudomonas syringae pv. tabaci induced hrp-independent HR in tomato

We have previously shown that flagellin of Pseudomonas syringae pv. tabaci is an elicitor that induces a hypersensitive reaction (HR) in nonhost tomato cells. Flagellin is the major HR elicitor produced by this pathogen, as shown by the inability of a flagellin-defective mutant, ΔfliC, to induce HR. Also, a ΔfliD mutant that secretes large amounts of monomer flagellins induces a strong HR in tomato. In this study, the possible involvement of an Hrp type III secretion system (TTSS) in flagellin-induced HR was investigated using flagella-defective mutants or Hrp TTSS-defective mutants. The hrcC gene encodes HrcC protein, which is required for Hrp pilus formation in the outer membrane. An hrcC mutation, introduced into the wild-type, ΔfliC, and ΔfliD mutants of P. syringae pv. tabaci did not affect swimming motility or flagellin secretion, whereas all ΔhrcC, ΔfliC, and ΔfliD mutants lost the ability to cause disease on host tobacco leaves. However, the ΔhrcC mutant and the ΔfliD/ΔhrcC double mutant were still able to induce HR cell death, expression of one of the defense-related genes hsr203J, and the generation of hydrogen peroxide in nonhost tomato cells. Thus, flagellin is required for both pathogenicity in host tobacco and HR in nonhost tomato. On the other hand, hrp TTSS is necessary for pathogenicity on host tobacco but is not indispensable to induce HR in nonhost tomato. These results clearly show that flagellin-induced HR is hrp-independent in tomato.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession number AB049570     

Defense responses of Arabidopsis thaliana inoculated with Pseudomonas syringae pv. tabaci wild type and defective mutants for flagellin (ΔfliC) and flagellin-glycosylation (Δorf1)

Flagellin, an essential component of the bacterial flagellar filament, is capable of inducing a hypersensitive response (HR), including cell death, in a nonhost plant. A flagellin-defective mutant (ΔfliC) of Pseudomonas syringae pv. tabaci lacks both the flagellar filament and motility, whereas a flagellin-glycosylation-defective mutant (Δorf1) retains the flagellar filament but lacks the glycosyl modification of flagellin protein. To investigate the role of flagellin protein and its glycosylation in the interaction with its nonhost Arabidopsis thaliana, we analyzed plant responses after inoculation with these bacteria. Inoculation with wild-type P. syringae pv. tabaci induced HR, with the generation of reactive oxygen species and cell death. In contrast, inoculation with either ΔfliC or Δorf1 mutant induced a low level of HR, and inoculated leaves developed a disease-like yellowing. These mutant bacteria multiplied better than the wild-type bacteria in A. thaliana. These results indicate that A. thaliana expresses a defense reaction in response to the bacterial flagellin with its glycosyl structure.     

Seasonal changes of azimuthal,radial, and tree-to-tree variations in sap flux affect stand transpiration estimates in a <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> forest,central Taiwan

Stand transpiration (E) estimated using the sap flux methods is affected by the azimuthal, radial, and tree-to-tree variations of sap flux. Although several studies have examined the relative importance of the three variations in estimating E, the seasonality of the three variations remains unknown. In the current study, we attempted to clarify whether the relative importance of these three variations could show seasonal changes. Using sap flux data measured in a subtropical cloud forest from August 2010 to July 2011, we calculated the differences resulting from omitting the three variations in estimating E. The effects of the three variations in estimating E showed seasonality. The azimuthal and tree-to-tree variations were more pronounced during winter, whereas the radial variation was more pronounced during summer. However, the effect of tree-to-tree variation was consistently much larger than the other two variations throughout the study period. The tree-to-tree variation is more important in estimating E monthly, seasonally and annually than both the azimuthal and radial variations, although all three variations have shown seasonality. In addition, the sensor allocation for summer would be acceptable for the practical estimation of E if aiming at the long time scale.     

<Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>-mediated transformation as a tool for random mutagenesis of <Emphasis Type="Italic">Colletotrichum trifolii</Emphasis>

We transformed Colletotrichum trifolii, the causal agent of alfalfa anthracnose, using Agrobacterium tumefaciens as a new tool for random insertional mutagenesis. Fungal spores of C. trifolii were transformed with T-DNA including the hygromycin phosphotransferase gene (hph). Southern analysis showed that every randomly selected transformant had a unique hybridization pattern of T-DNA, suggesting that the T-DNA was randomly integrated into the fungal genome. More significantly, about 75% of transformants had a single copy of the T-DNA. The results demonstrate that insertional mutagenesis via A. tumefaciens is a useful tool for studying the function of C. trifolii genes.     

Bacterial DNA activates immunity in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

To initiate defense responses against invasion of pathogenic organisms, animals and plants must recognize microbe-associated molecular patterns (MAMPs). In this study, the elicitor activity of bacterial DNA on the model plant Arabidopsis thaliana was examined. EcoRI-digested plasmid DNA induced defense responses such as generation of reactive oxygen species and deposition of callose, whereas SmaI- and HapII-digested plasmid DNA and EcoRI-digested herring DNA did not remarkably induce these responses. Further, methylation of the CpG sequence of plasmid DNA and Escherichia coli DNA reduced the level of the defense responses. The endocytosis inhibitors wortmannin and amantadine significantly inhibited DNA-induced defense responses. These results suggest that non-methylated CpG DNA, as a MAMP, induced defense responses in Arabidopsis and that non-methylated DNA seems to be translocated into the cytoplasm by endocytosis.     

Changes in daily rainfall characteristics in major rice producing provinces in Laos

In order to find the changes in rainfall characteristics and to examine the flood damage to lowland rice, a statistical analysis was carried out on the 40-year daily rainfall record at four rainfall stations (Thangon, Paksane, Seno, and Pakse) in Laos. The rainfall record was divided into two periods of 20 years each. The results of this analysis revealed the trends in changes regarding the frequency and the timing of the heavy daily rainfall events at the four stations. In Paksane in particular, heavy daily rainfall events tended to occur in the latter half of the rainy season.     

Hormone injections enhance the tolerance of land-locked ayu spermatozoa to cryopreservation

We evaluated the effects of maturation-stimulating hormones on the post-thaw motility of land-locked form ayu ( Plecoglossus altivelis ) spermatozoa. Male ayu were administered three intraperitoneal injections of either salmon pituitary extract (SPE; 0.2 or 0.6 mg g⁻¹ BW day⁻¹) or of 17, 20β-dihydroxy-4-pregnen-3-one (DHP; 2 or 10 μg g⁻¹ BW day⁻¹), the maturation-inducing steroid (MIS) in ayu. Before cryopreservation, the motility of spermatozoa of the SPE- and DHP-treated groups was significantly higher than that of the control group. Similarly, the comparative post-thaw motility (presented as a percentage of the motility obtained before cryopreservation) was significantly higher in the SPE group than in the control; however, there was no significant difference between the DHP group and the control. The effect of SPE and DHP on pre- and post-cryopreservation motility was not dose dependent. Our results suggest that the hormone(s) present in salmon pituitary are effective in enhancing the tolerance of ayu sperm cells to cryopreservation and that the MIS (DHP) is not involved in this process.     

Nitro-PAHs and PAHs in Atmospheric Particulate Matters and Sea Sediments in Hiroshima Bay Area, Japan

Atmospheric and sea sediment concentrations were measured for eight nitrated polycyclic aromatic hydrocarbons (nitro-PAHs) and three unsubstituted PAHs in a suburban area and sea sediments in the Hiroshima Bay watershed area, Japan, from July to December, 2006 (atmospheric particulate matter) and in September and November, 2004 (sea sediments). Atmospheric concentration was higher in winter than summer for both nitro-PAHs and PAHs. Concentrations in sea sediments were less than 10%, and pattern was similar to those of atmospheric particles. Several combustion emission sources were also measured, and the 1-NP/Pyr ratio was compared to environmental values. The ratio of atmospheric and sea sediments were significantly lower than diesel particulate matters. Further, the vehicle emission loading and sea sedimentation loading was evaluated in this watershed area, and from the comparison, the existence of other important sources PAHs were suggested.