SpringerActa Medica Okayama1345-26308562019A class III peroxidase PRX34 is a component of disease resistance in Arabidopsis405412ENLeiZhaoLaboratory of Plant Pathology and Genetic Engineering, Graduate School of Environmental and Life ScienceOkayama UniversityLe Thi PhuongLaboratory of Plant Pathology and Genetic Engineering, Graduate School of Environmental and Life ScienceOkayama UniversityMai Thanh LuanLaboratory of Plant Pathology and Genetic Engineering, Graduate School of Environmental and Life ScienceOkayama UniversityAprilia Nur FitriantiLaboratory of Plant Pathology and Genetic Engineering, Graduate School of Environmental and Life ScienceOkayama UniversityHidenoriMatsuiLaboratory of Plant Pathology and Genetic Engineering, Graduate School of Environmental and Life ScienceOkayama UniversityHirofumiNakagamiRIKEN Center for Sustainable Resource ScienceYoshiteruNoutoshiLaboratory of Plant Pathology and Genetic Engineering, Graduate School of Environmental and Life ScienceOkayama UniversityMikihiroYamamotoLaboratory of Plant Pathology and Genetic Engineering, Graduate School of Environmental and Life ScienceOkayama UniversityYukiIchinoseLaboratory of Plant Pathology and Genetic Engineering, Graduate School of Environmental and Life ScienceOkayama UniversityTomonoriShiraishiLaboratory of Plant Pathology and Genetic Engineering, Graduate School of Environmental and Life ScienceOkayama UniversityKazuhiroToyodaLaboratory of Plant Pathology and Genetic Engineering, Graduate School of Environmental and Life ScienceOkayama University PRX34 mediates the oxidative burst in Arabidopsis. Here we characterized two additional Arabidopsis prx34 null mutants (prx34-2, prx34-3), besides the well-studied prx34-1. Due to a decrease in corresponding peroxidase, the activity that generates reactive oxygen species (ROS) was significantly lower in cell wall extracts of prx34-2 and prx34-3 plants. Consistently, the prx34-2 and prx34-3 exhibited reduced accumulation both of ROS and callose in Flg22-elicitor-treated leaves, leading to enhanced susceptibility to bacterial and fungal pathogens. In contrast, ectopic expression of PRX34 in the wild type caused enhanced resistance. PRX34 is thus a component for disease resistance in Arabidopsis.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama2186-77551022013β-caryophylleneの植物に対する生育促進作用 および耐病性増進作用の解析714ENYasuoYamagiwaKazuhiroToyodaYoshishigeInagakiYukiIchinoseMitsuroHyakumachiaTomonoriShiraishiA plant growth-promoting fungus, Talaromyces wortmannii strain FS2 was isolated from an agricultural field at Okayama Pref. FS2 enhanced seed germination, root elongation and leaf growth of Brassica rapa var perviridis (Komatsuna). Such plant growth-promoting effect was observed in the same sealed chamber where FS2 was cultured on PDA medium separated from seedlings, suggesting effective volatile compound(s). GC‒MS analysis showed that FS2 emitted at least seven terpenoids, of which a volatile was identified as β‒caryophyllene. β‒caryophyllene alone promoted the growth of cucumber, Nicotiana benthamiana and barley. Furthermore β‒caryophyllene increased the yield of cucumber fruits. Interestingly, we found that β‒caryophyllene conditioned these plants to be resistant to respective diseases caused by Colletotrichum orbiculare, Botrytis cinerea or Blumeria graminis f. sp hordei. The findings indicate that β‒caryophyllene has desirable dual features and therefore, it is available to cultivation of many crops.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama2186-77551022013岡山県の栽培圃場における植物生育促進菌の探索と同定16ENYasuoYamagiwaKazuhiroToyodaYoshishigeInagakiYukiIchinoseTomonoriShiraishiA plant growth-promoting fungus was isolated from an agricultural field in Okayama Prefecture, Japan. The strain FS2, which enhanced seed germination, root elongation and leaf growth of Brassica rapa var. perviridis, was identified as Talaromyces wortmannii based on ITS1 sequence and its morphology.No potential conflict of interest relevant to this article was reported.Acta Medica Okayama27932008Modulation of defense signal transduction by flagellin-induced WRKY41 transcription factor in Arabidopsis thaliana303312ENKuniakiHigashiYasuhiroIshigaYoshishigeInagakiKazuhiroToyodaTomonoriShiraishiYukiIchinose<p>Flagellin, a component of the flagellar filament of Pseudomonas syringae pv. tabaci 6605 (Pta), induces hypersensitive reaction in its non-host Arabidopsis thaliana. We identified the WRKY41 gene, which belongs to a multigene family encoding WRKY plant-specific transcription factors, as one of the flagellin-inducible genes in A. thaliana. Expression of WRKY41 is induced by inoculation with the incompatible pathogen P. syringae pv. tomato DC3000 (Pto) possessing AvrRpt2 and the non-host pathogens Pta within 6-h after inoculation, but not by inoculation with the compatible Pto. Expression of WRKY41 was also induced by inoculation of A. thaliana with an hrp-type three secretion system (T3SS)-defective mutant of Pto, indicating that effectors produced by T3SS in the Pto wild-type suppress the activation of WRKY41. Arabidopsis overexpressing WRKY41 showed enhanced resistance to the Pto wild-type but increased susceptibility to Erwinia carotovora EC1. WRKY41-overexpressing Arabidopsis constitutively expresses the PR5 gene, but suppresses the methyl jasmonate-induced PDF1.2 gene expression. These results demonstrate that WRKY41 may be a key regulator in the cross talk of salicylic acid and jasmonic acid pathways.</p>
No potential conflict of interest relevant to this article was reported.SpringerActa Medica Okayama27942008Gac two-component system in Pseudomonas syringae pv. tabaci is required for virulence but not for hypersensitive reaction313322ENMizuriMarutaniFumikoTaguchiYujiroOgawaMijan Md.HossainInagakiYoshishigeKazuhiroToyodaTomonoriShiraishiYukiIchinose<p>Pseudomonas syringae pv. tabaci 6605 causes wildfire disease on host tobacco plants. To investigate the regulatory mechanism of the expression of virulence, Gac two-Component system-defective mutants, Delta gacA and Delta gacS, and a double mutant, Delta gacA Delta gacS, were generated. These mutants produced smaller amounts of N-acyl homoserine lactones required for quorum sensing, had lost swarming motility, and had reduced expression of virulence-related hrp genes and the algT gene required for exopolysaccharide production. The ability of the mutants to cause disease symptoms in their host tobacco plant was remarkably reduced, while they retained the ability to induce hypersensitive reaction (HR) in the nonhost plants. These results indicated that the Gac two-component system of P. syringae pv. tabaci 6605 is indispensable for virulence on the host plant, but not for HR induction in the nonhost plants.</p>No potential conflict of interest relevant to this article was reported.Acta Medica Okayama1822007Elicitin-responsive lectin-like receptor kinase genes in BY-2 cells152159ENMichikoSasabeKanaNaitoHirokoSuenagaTakakoIkedaKazuhiroToyodaYoshishigeInagakiTomonoriShiraishiYukiIchinose<p>The inhibition of elicitor-induced plant defense responses by the protein kinase inhibitors K252a and staurosporine indicates that defense responses require protein phosphorylation. We isolated a cDNA clone encoding Nicotiana tabacum lectin-like receptor protein kinase 1 ( NtlecRK1), an elicitor-responsive gene; in tobacco bright yellow ( BY-2) cells by a differential display method. NtlecRK forms a gene family with at least three members in tobacco. All three NtlecRK genes potentially encode the N-terminal legume lectin domain, transmembrane domain and C-terminal Ser/Thr-type protein kinase domain. Green fluorescent protein ( GFP) fusion showed that the NtlecRK1 protein was located on the plasma membrane. In addition, NtlecRK1 and 3 were responsive to INF1 elicitin and the bacterial elicitor harpin. These results indicate that NtlecRKs are membrane-located protein kinases that are induced during defense responses in BY-2 cells.</p>
No potential conflict of interest relevant to this article was reported.American Society for MicrobiologyActa Medica Okayama0021-9193188242006A homologue of the 3-oxoacyl-(acyl carrier protein) synthase III gene located in the glycosylation island of Pseudomonas syringae pv. tabaci regulates virulence factors via N-acyl homoserine lactone and fatty acid synthesis83768384ENFumikoTaguchiYujiroOgawaKasumiTakeuchiTomokoSuzukiKazuhiroToyodaTomonoriShiraishiYukiIchinosePseudomonas syringae pv. tabaci 6605 possesses a genetic region involved in flagellin glycosylation. This region is composed of three open reading frames: orf1, orf2, and orf3. Our previous study revealed that orf1 and orf2 encode glycosyltransferases; on the other hand, orf3 has no role in posttranslational modification of flagellin. Although the function of Orf3 remained unclear, an orf3 deletion mutant (Delta orf3 mutant) had reduced virulence on tobacco plants. Orf3 shows significant homology to a 3-oxoacyl-(acyl carrier protein) synthase III in the fatty acid elongation cycle. The Delta orf3 mutant had a significantly reduced ability to form acyl homoserine lactones (AHLs), which are quorum-sensing molecules, suggesting that Orf3 is required for AHL synthesis. In comparison with the wild-type strain, swarming motility, biosurfactant production, and tolerance to H2O2 and antibiotics were enhanced in the Delta orf3 mutant. A scanning electron micrograph of inoculated bacteria on the tobacco leaf surface revealed that there is little extracellular polymeric substance matrix surrounding the cells in the Delta orf3 mutant. The phenotypes of the Delta orf3 mutant and an AHL synthesis (Delta psyI) mutant were similar, although the mutant-specific characteristics were more extreme in the Delta orf3 mutant. The swarming motility of the Delta orf3 mutant was greater than that of the Delta psyI mutant. This was attributed to the synergistic effects of the overproduction of biosurfactants and/or alternative fatty acid metabolism in the Delta orf3 mutant. Furthermore, the amounts of iron and biosurfactant seem to be involved in biofilm development under quorum-sensing regulation in P. syringae pv. tabaci 6605.No potential conflict of interest relevant to this article was reported.Blackwell PublishingActa Medica Okayama1462-5814862006Identification of glycosylation genes and glycosylated amino acids of flagellin in Pseudomonas syringae pv. tabaci923938ENFumikoTaguchiKasumiTakeuchiEtsukoKatohKatsuyoshiMurataTomokoSuzukiMizuriMarutaniTakayukiKawasakiMinakoEguchiShizueKatohHanaekakuChihiroYasudaYoshishigeInagakiKazuhiroToyodaTomonoriShiraishiYukiIchinoseA glycosylation island is a genetic region required for glycosylation. The glycosylation island of flagellin in Pseudomonas syringae pv. tabaci 6605 consists of three orfs: orf1, orf2 and orf3. Orf1 and orf2 encode putative glycosyltransferases, and their deletion mutants, Delta orf1 and Delta orf2, exhibit deficient flagellin glycosylation or produce partially glycosylated flagellin respectively. Digestion of glycosylated flagellin from wild-type bacteria and non-glycosylated flagellin from Delta orf1 mutant using aspartic N-peptidase and subsequent HPLC analysis revealed candidate glycosylated amino acids. By generation of site-directed Ser/Ala-substituted mutants, all glycosylated amino acid residues were identified at positions 143, 164, 176, 183, 193 and 201. Matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS) analysis revealed that each glycan was about 540 Da. While all glycosylation-defective mutants retained swimming ability, swarming ability was reduced in the Delta orf1, Delta orf2 and Ser/Ala-substituted mutants. All glycosylation mutants were also found to be impaired in the ability to adhere to a polystyrene surface and in the ability to cause disease in tobacco. Based on the predicted tertiary structure of flagellin, S176 and S183 are expected to be located on most external surface of the flagellum. Thus the effect of Ala-substitution of these serines is stronger than that of other serines. These results suggest that glycosylation of flagellin in P. syringae pv. tabaci 6605 is required for bacterial virulence. It is also possible that glycosylation of flagellin may mask elicitor function of flagellin molecule.No potential conflict of interest relevant to this article was reported.Acta Medica Okayama842007Suppression of Cdc27B expression induces plant defence responses365373ENChikakoKudoTomokoSuzukiSumieFukuokaShutaAsaiHirokoSuenagaMichikoSasabeYoshitakaTakanoTetsuroOkunoKazuhiroToyodaTomonoriShiraishiYukiIchinoseYoshi-ShigeInagaki<p>Non-host resistance is the most general form of disease resistance in plants because it is effective against most phytopathogens. The importance of hypersensitive responses (HRs) in non-host resistance of Nicotiana species to the oomycete Phytophthora is clear. INF1 elicitin, an elicitor obtained from the late-blight pathogen Phytophthora infestans, is sufficient to induce a typical HR in Nicotiana species. The molecular mechanisms that underlie the non-host resistance component of plant defence responses have been investigated using differential-display polymerase chain reaction (PCR) in a model HR system between INF1 elicitin and tobacco BY-2 cells. Differential-display PCR has revealed that Cdc27B is down-regulated in tobacco BY- 2 cells after treatment with INF1 elicitin. Cdc27B is one of 13 essential components of the anaphase- promoting complex or cyclosome ( APC/ C)-type E3 ubiquitin ligase complex in yeast. This APC/C-type E3 ubiquitin ligase complex regulates G2-to-M phase transition of the cell cycle by proteolytic degradation. In this study, we investigated the roles of this gene, NbCdc27B, in plant defence responses using virus-induced gene silencing. Suppression of NbCdc27B in Nicotiana benthamiana plants induced defence responses and a gain of resistance to Colletotrichum lagenarium fungus. Elicitin-induced hypersensitive cell death (HCD) was inhibited mildly in plants silenced with tobacco rattle virus:: Cdc27B. Cdc27B could manage the signalling pathways of plant defence responses as a negative regulator without HCD.</p>No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-0254 9912010病原菌の感染行動に及ぼす FFC セラミック水の効果について2734ENTomonoriShiraishiKazuhiroToyodaTomokoSuzukiAkaneMeguroSachikoHasegawaTomioNishimuraHitoshiKunohIn this report, an effect of FFC-ceramic (FFC-Japan Co. Ltd., Tsu) water on the process of infection by a pea fungal pathogen, Mycosphaerella pinodes was investigated. Energy dispersive X-ray analysis showed that both of the FFC-ceramic water and a common ceramic water contained mainly Ca and S elements, of which the relative atomic percentages were 53~56% and 44~45%, respectively. Lesion formation by pycnospores of M. pinodes on pea leaves was inhibited severely by the application with both ceramic waters at the 1/2~1/6 concentration of saturated solution. Cytological observation under microscope showed that germination, germ-tube elongation and penetration were severely inhibited by these ceramic waters. However, such inhibitory effect of FFC-ceramic water was superior to that of the common ceramic water. On ethanol-killed pea epidermal tissues, both FFC-ceramic water and the common ceramic water blocked the germination, germ-tube elongation and penetration by the pathogen, indicating
the direct effect of both ceramic waters on the fungus. In this case, the inhibiting effect of FFC ceramic water was more intensive than the common ceramic water. CaSO(4) at a 1/2~1/4 concentration of saturated solution blocked penetration by the fungus on the killed epidermis of onion bulb but scarcely affected germination and germ-tube elongation. Based on these results, we discussed the role of FFC-ceramic water in disease tolerance of plants and its availability for cultivation.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-0254 9912010FFC セラミック水(TM)による植物アピラーゼの活性化作用2126ENKazuhiroToyodaSachikoMatsuokaAkaneMeguroSachikoHasegawaTomioNishimuraHitoshiKunohTomonoriShiraishiThe FFC ceramics(TM) from FFC Japan Co., Ltd. are now widely used in the fields of agriculture, fishery and food industry in Japan. Recently the FFC ceramic beads-based technology has been also applied to meet several environmental problems including pollution in sea, lakes and rivers. In this study the FFC ceramic water was tested for effect on plant enzyme, potato apyrase (EC 3.6.1.5; ATP-diphosphohydrolase), which hydrolyses nucleoside triphosphate (NTP) and -diphosphate (NDP) to produce corresponding nucleoside monophosphate (NMP) and inorganic phosphate (Pi). Addition of the FFC ceramic water to the enzyme reaction mixture markedly enhanced ATP-hydrolyzing activity, when used as ATP as substrate. However, the concomitant presence of Ca(2+) chelator, EGTA (O,O'-bis(2-aminoethyl)ethyleneglycol-N,N,N',N'-tetraacetic acid) with the FFC ceramic water, completely abolished the enzyme activation. In fact, exogenous calcium ion such as CaSO4 mimicked the FFC ceramic water. These results indicate that apyrase activation by the FFC ceramic water largely depends on calcium ions. On the other hand, when the FFC ceramic water prepared from "used" ceramics was tested for the apyrase activity, the enhanced effect on apyrase was decreased compared to the FFC ceramic water from "new" ones. This result, consistent with our present data covering concentration of calcium ions and conductivity, indicates that long and/or successive usage of the ceramic beads results in decrease of contents of released minerals, especially calcium ions. The apyrase-based enzyme assay presented here is probably applicable to estimate and quantify the effect of FFC ceramic water.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-0254 9912010Enhancement of Growth and Yield of Barley by the Soil Conditioner FFC-ace1320ENKeikoFujitaTomokoSuzukiSachikoHasegawaAkaneMeguroHiroyukiSugiuraKazuhiroToyodaTomonoriShiraishiEiSakaguchiTomioNishimuraHitoshiKunohThe effects of a unique soil conditioner, FFC-ace, on photosynthesis, transpiration, growth and yield of barley were examined in a field experiment. FFC-ace well-mixed with sandy soil greatly enhanced root and shoot growth, tillering and the number of grains per stock. The total yield in the treated plot increased by about 172%. The plants grown in the FFC-ace plot were greener and contained a higher level of chlorophyll, compared with the control. Photosynthesis and transpiration, which are tightly linked to productivity were also significantly enhanced at the broad range of photon flux observed in our study. The quality of grain harvested from the FFC-ace plot was similar to the control plot in terms of nutritional and inorganic components. The increased photosynthesis in the FFC-ace treated barley reflects a higher absorption of CO(2) from the atmosphere. It was also noted that the efficiency of water utilization for photosynthesis was significantly greater under the high light intensity in the treated plot. The relationship between application of FFC-ace and absorption of atmospheric CO(2) is discussed. Our investigation provides
data showing that application of FFC-ace to soil significantly reduces water requirements for plant growth and yield.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02548611997Orthovanadate Induces Phytoalexin Production in Pea Suspension-Cultured Cells3341ENTomoharuKawaharaKazuhiroToyodaAkinoriKibaYukiIchinoseTetsujiYamadaTomonoriShiraishiWe previously reported that the addition of orthovanadate suppressed the defense responses of plant differentiated tissues induced by a fungal elicitor.In this report,the effect of orthovanadate on the defense response of pea cultured cells was examined.The activities of ATPase and PI metabolism in plasma membrance fraction,which was prepared from suspension-cultured cells,were inhibited in vitro by orthovanadate as well as those in plasma membrances from pea epicotyl tissues. However,orthovanadate alone induced the accumulation of a phytoalexin, pisatin in suspention-clutured cells of pea in a manner similar to CuCl2.The viability of pea suspension-cultured cells was decreased by orthovanadate as well as by CuCl2.These results indicated that orthovanadate acts as an abiotic elicitor to pea suspension-cultured cell as observed in those of red bean,peanut and Perunia hybrida.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02548711998Phosphorylation of Phosphatidylinositols and Production of Ｌｙsophospholipid in Pea Plasma Membrane Are Coordinately Regulated by Elicitor and Suppressor from Mycosphaerella pinodes
109116ENKazuhiroToyodaMasashiKoyamaRumiMizukoshiYukiIchinoseTetsujiYamadaTomonoriShiraishiEffects of elicitor and suppressor from a pea pathogen, Mycosphaerella pinodes, on Pl etabolism in pea plasma membrane were examined in vitro. The elicitor induced rapid phosphorylation of phosphatidylinositols as well as production of lysophospholipid in plasma membranes, but these responses were severely inhibited by the suppressor. These results indicate that a membrane-associated phospholipase A is regulated coordinately by fungal signals, together with Pl metabolism, and that it may participate in signal transduction pathways leading to defense responses. To evaluate a possible rote of phospholipase A activation in induction of a pea defense response, the effect of free fatty acid on induction of a phytoalexin accumulation was also examined. When pea leaves were treated with linoleic- or linolenic acid, most commonly released in plant cells by phospholipase A, the accumulation of pisatin was induced even in the absence of the elicitor. It is, therefore, conceivable that free fatty acid(s) released from plasma membrane is also implicated in the early stage of elicitor-signal transduction in pea.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02548711998Transformation of Mutualistic Fungal Acremonium Endophyte99107ENAhamadYunusYukiIchinoseTomonoriShiraishiTetsujiYamadaConditions have been developed for transforming protoplasts of the Acremonium endophyte by PEG 4000 and electroporation. Transformation by PEG exhibited a higher number of transformants than by electroporation. lntegration of iaaM gene into the genome was examined by PCR and Southern blot hybridization analysis. PCR product showed that transformants banded at around 1.7 kb corresponding to the size of iaaM gene. Hybridization of the digests of genomic DNA with iaaM gene as DNA probe showed that the number of hybridized band signals was different between transformant and non-transformant. These results might indicate that PEG is an effective method for the transformation of Acremonium endophyte and that there are repeated copies of the iaaM homologous sequences in the genome of Acremonium.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02548711998Cis-Regulatory Elements and Trans-acting Factors Involved in the Activation of a Member of Elicitor-Responsive Pea Chalcone Synthase Gene Family, PSCHS29197ENYukiIchinoseMasayukiItoHikaruSekiKazuhiroToyodaTomonoriShiraishiTetsujiYamadaTo elucidate the elicitor-mediated transcriptional activation in one of the chalcone synthase genes, PSCHS2 in pea, we investigated the putative cis-regulatory elements in the promoter sequence and trans-acting DNA binding proteins. The promoter up to -471 from the transcription start site of PSCHS2 gave considerable level of basal transcriptional activity. Nuclear extract from elicitortreated pea epicotyls formed DNA-protein complexes with three independent DNA fragments spanning from +83 to -484 of PSCHS2 with low mobility (LMC,low mobility complex) in the gel mobility shift assay. Since the LMC formation was blocked by the treatment of nuclear extract with alkaline phosphatase, the phosphorylation of some nuclear factor(s) assists LMC formation. These results indicate that the bindings of the putative positive nuclear factors to the multiple cisregulatory elements in PSCHS2 promoter region were enhanced by elicitortreatment that might result in transcriptional activation.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02548811999Co-purification of Plasma Membrane ATPase and Phosphatidylinositol Kinase from Pea Plasma Membranes3138ENKazuhiroToyodaYukiIchinoseTetsujiYamadaTomonoriShiraishiThe plasma membrance ATPase was partially purified by a linear glycerol density gradient centrifugation of the detergent-solubilized plasma membrance poteins and subsequent separation by a size-exclusion column chromatogrphy. A purified
ATPase preparation is shown to contain a 97.6kDa protein that was cross-reacted with an antibody raised against mung bean H+-ATPase. The preparation also exhibited the phosphorylation of exogenous phosphatidylionsitol(PI) when supplized with [γ-32P]ATP. These results indicate that one form plasma membrance ATPase is co-purified with PI kinase.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02548811999Molecular Cloning of a cDNA Encoding a Putative DNA-Binding Zinc-Finger Protein in Pea2530ENYukiIchinoseAiEndohShirohSanematsuKazuhiroToyodaTomonoriShiraishiTetsujiYamadaWe constructed cDNA library from pea epicotyls treated with fungal elicitor for 5hrs, and performed differential screening using the individual 32P-cDNA probe derived from poly(A)+ RNA prepared from elicitor- and water-treated epicotyls. As a result of the screening, we have isolated from elicitor- and water-treated epicotyls. As a results of the screening, we have isolated about 90 cDNA clones as candidates for elicitor-inducible genes, and their nucleotide sequences have been partially determined. One of these clones, E31 was a pea homolog of the putative zinc-finger proteins, Ljpzf in Lotus japonicus and Gmpz in soybean. E31 possesses 1,726bp insert, and encodes an open reading frame corresponding to position 82 amino acids from N-terminus to the C-terminal end in Ljpzf. The protein product of E31 was designated as Pspzf. Pspzf also possesses nuclear localization signal(NLS), HKRK, and Cys3His2Cys3(RIngH2) motif at the same position to LjpZF and putative C-terminal end of the deduced amino acid sequences, respectively. Since zinc-finger motif is one of the well-known DNA-binding domains, Ljpzf and Pspzf might be able to bins to a particular DNA sequence and regulate transcriptional activity in plants.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02549212003Possible Involvement of AAAG Motif and PsDof1 in Elicitor-Induced Gene Expression in Pea2126ENHikaruSekiMizuriMarutaniYoshishigeInagakiKazuhiroToyodaTomonoriShiraishiYukiIchinoseRecently, we, isolated cDNA clone, PsDof1, from clicitor-treated pea cDNA library. The putative gene product, a PsDof1, encodes DNA binding protein that specifically binds the DNA fragment containing AAAG core sequence. In this paper we report that GST-PsDof1 fusion protein specifically binds to the promoter region containing AAAG core sequence(s) of PsCHS1, one of the elicitor-inducible genes encoding chalcone synthase (CHS). Furthermore the addition of DNA fragment containing AAAG motif to the 35S minimal promoter provided the elicitor-responsibility in transient transfection assay using pea protoplasts. These results suggest that PsDof1 might be involved in defense responses by acitivating the transcription by a binding to AAAG core sequence in the promoter of the defense-related genes in pea.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02548011992A Reporter Gene Expression Under the Control of a Pea Phenylalanine Ammonia Lyase-gene Promoter5160ENTetsujiYamadaTadaakiHashimotoPermpongSriprasertsakHisaharuKatoYukiIchinoseTomonoriShiraishiHachiroOkuHigh yields of viable pea protoplasts were produced from suspension cultured cells derived from calli formed from embryogenic tissues or leaves and the conditions for the optimum expression of chloramphenicol acetyltransferase (CAT) fused to the phenylalanine ammonia-lyase gene of Pisum sativum (pPAL1-15) were investigated by transient assay after electroporation. A fungal elicitor isolated from a pea pathogen, Mycosphaerella pinodes, and the reduced from of glutathione induced the expression of PAL promoter but orthovanadate, a plasma membrane ATPase inhibitor, considerably suppressed the gene expression. Rice protoplasts were also prepared from the suspension cultured cells derived from embryonic tissues, and the effects of elicitors on the expression of CAT in pPAL1-15-electroporated rice protoplasts were examined. No distinctive induction of CAT activity was observed by the treatment of rice protoplasts with a chitosan oligomer elicitor.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02545911982Isolation and Partial Characterization of an Elicitor of Pisatin Production from Spore Germination Fluid of Pea Pathogen, Mycosphaerella pinodes19ENPorntipThanutongHachiroOkuTomonoriShiraishiSeijiOuchiAn elicitor of pisatin accumulation was isolated and partially characterized from spore germination fluid, mycelial extract, and cell wall preparation of a pea pathogen, Mycosphaerella pinodes. The elicitor was found to be a polysaccharide or glycoprotein and the active component was composed of glucose. The approximate molecular weight was 70,000 daltons. This elicitor elicited pisatin accumulation in pea leaves at a concentration of 10 ug/ml and the maximum activity was ca. 350-400 ug/ml, but did not elicit phytoalexins in soybean, bean, and red clover. High molecular weight preparations from the mycelial extract and mycelial wall of Mycosphaerella melonis and Stemphylium sarcinaeforme, nonpathogens of pea, elicited pisatin accumulation in pea leaves.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02545411979Effect of Fungicides, Benomyl and Thiram on Soil Microflora and Some Inhabitant Fungi18ENHachiroOkuKazuoOkiTomonoriShiraishiKenjiSatoSeijiOuchiEffects of Benomyl and Thiram on the soil microflora and some soil inhabitant fungi were studied using the_ soil (sandy loam) of the experimental field, Okayama University. Under field conditions, heavy application of Benomyl did not affect significantly the soil microflora. Thiram, however, reduced the fungal population in soil to 1/6 at the next day of treatment, but recovered to the normal level after 6 days. Under laboratory conditions, both fungicides did not affect soil microflora. Population of Benomyl-to lerant fungi was about 1/10 of the total fungi and increased slightly in the field soil by treatment with Benomyl at the later stage of experiment during June to October. Neither Thiram-tolerant fungus nor bacterium was found in both Thiram-treated and non-treated soils. A fungus highly tolerant to Benomyl was isolated and identified as Aspergillus versicolor, and found to not have the metabolic activity to degrade BCM. The absorption of BCM by the mycelia of this tolerant fungus, A. versicolor, was less than half of the BCM-sensitive one, such as Cladosporium harbarum.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02544911977ブドウ褐点病の防除について16ENHachiroOkuMotomuHatamotoSeijiOuchiTomonoriShiraishiMichihiroTateishiShintaroFujiiブドウ褐点病防除のための農薬のスクリーニングを行なうとともに,発生生態の観察結果を含めて本病の防除施策について論じた. 供用農薬のなかでは,ベンレート,サニパー,DF125が有効であった. 自然発生,特に汚果の防止には硫酸ニコチンの混用が有効であった. 外気の直接導入を避けるような温室出入口の構造改善,室内浄化,誘枝線の更新,過湿の回避などが有効な技術的な防除策と考えられた。No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02544811976Brown Spot of Grapes Caused by Cladosporium cladosporioides and Cladosporium herbarum1722ENSeijiOuchiMotomuHatamotoHachiroOkuTomonoriShiraishiTatsuoYokoyamaMichihiroTateishiShintaroFujiiCladosporium cladosporioides (Fresenius) de Vries and Cladosporium herbarum (Pers. ) Link ex Fr. were isolated from grape berries that had been commonly called 'black navel', and were found to cause the disease. In view of the colour of lesions that were observed most frequently at the stage of maturing, the disease was named brown spot of grapes.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02544711976オオムギうどんこ病菌吸器細胞壁におけるキチン成分について2124ENTomonoriShiraishiSeijiOuchiHachiroOku純寄生性病害の感染成立に中心的役割を演ずると考えられている吸器の性質を明らかにするために,オオムギうどんこ病菌の吸器形成過程における吸器壁のキチン質の生成に関して組織化学的に検討した. その結果,吸器壁はキトーサン反応陽性で,菌糸壁と同様キチン質がその骨格をなしているものと考えられる. さらに,経時的な検討の結果,感染初期に形成される吸器原基において,すでにキチン反応が陽性であり,キチン合成系の活性化は,吸器機能発現のための一つの重要な過程であると推定される。No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02544711976Uptake and Dechlorination of an Organochlorine Fungicide, Tetrachloroisophthalonitrile, Daconil, by Some Soil Fungi713ENHachiroOkuTomonoriShiraishiSeijiOuchiSayokoHoriuchiFungi tolerant to an organochlorine fungicide, tetrachloroisophthalonitrile (TCPN), was selected and isolated from various soil samples, and examined for the activity to metabolize TCPN. The population of tolerant fungi was extremely large in the plastic house - soil on which TCPN has been used for six years to control diseases. This is probably due to the change of soil microflora owing to the decline of TCPN-sensitive competitors. Many tolerant fungi took up TCPN very rapidly from culture filtrate. Of ten tolerant isolates tested, two isolates, C-F- I and V-P-5, were found to metabolize TCPN. The metabolite was isolated in pure form by preparative TLC, and identified as trichloroisophthalonitrile by mass spectrometry. C-F-1 was identified as Aspergillus luchuensis and V-P-5 as Penicillium godlewskii. Namely, these soil fungi dechlorinate from tetrachloroisophthalontrile to trichloroisophthalonitrile.No potential conflict of interest relevant to this article was reported.岡山大学農学部Acta Medica Okayama0474-02544211973Phytoalexin Induction by Some Agricutural Fungicides and Phytotoxic Metabolites of Pathogenic Fungi1720ENHachiroOkuToshiroNakanishiTomonoriShiraishiSeijiOuchiThe endocarp of the fresh pea pod incubated with solution or suspension of agricultural fungicides, phytotoxic metabolites of plant pathogenic fungi formed the pea phytoalexin, pisatin. Among the compounds tested, cycloheximide, triazine, dichlone, phenyimercury acetate, UV degradation product of phenylmercury acetate, triphenyltin fungicides, and 3-hydroxy-5-methylisoxazole induced pisatin. Ophiobolin, a toxin from Cochliobolus mtyabeanus, and ascochitine, a toxic metabolite from Ascochyta fabae, also induced pisatin. The possibilities of the development of harmless plant disease control agents was discussed in relation- to the induced synthesis of phytoalexins.No potential conflict of interest relevant to this article was reported.