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1.
Akinori Kiba Takako Ohgawara Kazuhiro Toyoda Miho Inoue-Ozaki Tadahiro Takeda Uppalapati Srinivasa Rao Toshiaki Kato Yuki Ichinose Tomonori Shiraishi 《Journal of General Plant Pathology》2006,72(4):228-237
In the plant cell wall of Pisum sativum seedlings, we found an NTPase (E.C. 3.6.1.5.) with ATP-hydrolyzing activity that was regulated by an elicitor and suppressors
of defense from pea pathogen Mycosphaerella pinodes. The ATPase-rich fraction was purified from pea cell walls by NaCl solubilization, ammonium sulfate precipitation, and chromatography
with an ATP-conjugated agarose column and an anion-exchange column. The specific activity of the final ATPase-rich fraction
increased 600-fold over that of the initial NaCl-solubilized fraction. The purified ATPase-rich fraction also had peroxidase
activity and generated superoxide, both of which were regulated by the M. pinodes elicitor and suppressor (supprescins). Active staining and Western blot analysis also showed that the ATPase was copurified
along with peroxidases. In this fraction, a biotinylated elicitor and the supprescins were bound primarily and specifically
to ca. 55-kDa protein (CWP-55) with an N-terminal amino acid sequence of QEEISSYAVVFDA. The cDNA clone of CWP-55 contained five ACR domains, which are conserved in
the apyrases (NTPases), and the protein is identical to a pea NTPase cDNA (GenBank accession AB071369). Based on these results, we discuss a role for the plant cell wall in recognizing exogenous
signal molecules. 相似文献
2.
Tomoharu Kawahara Kazuhiro Toyoda Akinori Kiba Akiko Miura Takako Ohgawara Mikihiro Yamamoto Yoshishige Inagaki Yuki Ichinose Tomonori Shiraishi 《Journal of General Plant Pathology》2003,69(1):33-38
Two nucleoside triphosphatase (NTPase) cDNA clones were isolated from a cDNA library of Pisum sativum L., cv. Midoriusui. The genes encoding the cDNAs were designated PsAPY1 and PsAPY2. PsAPY1 included the N-terminal amino acid sequence of an NTPase bound to pea cell wall. The phylogenic analysis indicated that PsAPY1 belongs to an NTPase subfamily responsive to environmental stimuli and that PsAPY2 belongs to a discrete subfamily, the physiological role of which is almost unknown. The adenosine triphosphatase activity
of recombinant PsAPY1 was regulated by an elicitor and a suppressor from the pea pathogen Mycosphaerella pinodes. Based on these findings, we discuss the role of NTPases in response to biological stresses.
Received: May 27, 2002 / Accepted: July 31, 2002 相似文献
3.
Tomonari Kasai Tomoko Suzuki Kozue Ono Ken'ichi Ogawa Yoshishige Inagaki Yuki Ichinose Kazuhiro Toyoda Tomonori Shiraishi 《Journal of General Plant Pathology》2006,72(5):265-272
We previously reported that the release of O2
− from isolated pea cell walls was enhanced by a 70-kDa glycoprotein elicitor but was suppressed by mucin-type glycopeptide
suppressors (supprescins A and B) prepared from pycnospore germination fluid of Mycosphaerella pinodes, causal agent of Mycosphaerella blight of pea. Here, we show that superoxide dismutase (SOD) in the apoplast fluid/cell wall
of pea seedlings responds to the fungal elicitor and suppressor molecules. In a pharmacological study and with internal amino
acid sequencing, the apoplastic SOD in a pea cultivar Midoriusui was found to be a Cu/Zn type SOD. We cloned a full-length
cDNA of the Cu/Zn-SOD and designated it as PsCu/Zn-SOD1. An increase in PsCu/Zn-SOD1 mRNA and the PsCu/Zn-SOD1 protein was induced by treatment with the elicitor more intensively
than by wounding. Such induction by the elicitor or wounding, however, was inhibited by the concomitant presence of supprescins.
The SOD activity of recombinant PsCu/Zn-SOD1 was regulated directly by these signal molecules in a manner similar to their
effect on the SOD activity in the apoplastic fluid and in the cell wall-bound proteins. Based on these findings, we discuss
a role for PsCu/Zn-SOD1 in the pea defense response.
The nucleotide sequence data of PsCu/Zn-SOD1 reported are available in the DDBJ/EMBL/GenBank databases under accession number AB189165. 相似文献
4.
Hirotaka Takahashi Kazuhiro Toyoda Yuzo Hirakawa Kunihiko Morishita Toshiaki Kato Yoshishige Inagaki Yuki Ichinose Tomonori Shiraishi 《Journal of General Plant Pathology》2006,72(3):143-151
Apyrases (NTPases) are associated with both compatible and incompatible interactions between plants and microorganisms. Previously
we reported that the ATPase activities of cell-wall-bound apyrases of several leguminous plants, such as pea, cowpea, soybean,
and kidney bean, were enhanced by a glycoprotein elicitor and were inhibited in a species-specific manner by mucin-type glycopeptide
suppressors secreted from a pea pathogenic fungus, Mycosphaerella pinodes. In this study, we isolated two apyrase genes, VsNTPase1 and VsNTPase2, from a cDNA library of Vigna sinensis Endl. cv. Sanjakusasage. Based on phylogenetic analysis, VsNTPase1 may belong to a group that responds to environmental stimuli.
In a transient assay using DNA bombardment, a fusion protein of green fluorescent protein (GFP) and the N-terminal putative signal sequence of VsNTPase1 was distributed in the nucleus, cytoplasm (cytoskeletal structure), and cell
wall. On the other hand, a fusion protein of GFP and the N-terminal putative VsNTPase2-signal sequence was localized in the cytoplasm, especially in small particles (perhaps mitochondria).
A recombinant VsNTPase1 expressed in Spodoptera frugiperda 21 cells responded directly to signal molecules from several phytopathogenic microorganisms. Here, we discuss the role of
apyrases in recognizing and responding to exogenous signals.
The nucleotide sequences of VsNTPase1 and VsNTPase2 in this article have been submitted to DDBJ as accession numbers AB196769
and AB196770, respectively. 相似文献
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7.
Rui TANAKA Fumiko TAGUCHI Yuki ICHINOSE Kazuhiro TOYODA Tomonori SHIRAISHI Tetsuji YAMADA 《Journal of General Plant Pathology》2001,67(2):148-151
To investigate the role of the proteinaceous elicitor, harpin, on host and nonhost plants, we isolated the harpin-coding gene,
hrpZ, from Pseudomonas syringae pvs. pisi, glycinea, tabaci and tomato. Effects of the recombinant harpin proteins on pea plants were analyzed and compared with the effects of the corresponding
bacterial treatment. After inoculation of pea with pea pathogen P. syringae pv. pisi, the bacterial population increased and the accumulation of PAL-mRNA and pisatin was inhibited. The nonpathogenic pathovars, glycinea, tabaci and tomato induced both defense responses in pea. However, none of the harpins induced the hypersensitive reaction or accumulation of
PAL-mRNA and pisatin in pea. Harpins from P. syringae pvs. glycinea, tomato and pisi did induce these defense responses in tobacco, however, suggesting that externally applied harpins either are not recognized
or are nonfunctional in pea plants.
Received 27 June 2000/ Accepted in revised form 21 February 2001 相似文献
8.
ABSTRACT Fractions solubilized with NaCl from cell walls of pea and cowpea plants catalyzed the formation of blue formazan from nitroblue tetrazolium. Because superoxide dismutase decreased formazan production by over 90%, superoxide anion (O(2) ) may participate in the formation of formazan in the solubilized cell wall fractions. The formazan formation in the fractions solubilized from pea and cowpea cell walls was markedly reduced by exclusion of NAD(P)H, manganese ion, or p-coumaric acid from the reaction mixture. The formazan formation was severely inhibited by salicylhydroxamic acid and catalase, but not by imidazole, pyridine, quinacrine, and diphenyleneiodonium. An elicitor preparation from the pea pathogen Mycosphaerella pinodes enhanced the activities of formazan formation nonspecifically in both pea and cowpea fractions. The suppressor preparation from M. pinodes inhibited the activity in the pea fraction in the presence or absence of the elicitor. In the cowpea fraction, however, the suppressor did not inhibit the elicitor-enhanced activity, and the suppressor alone stimulated formazan formation. These results indicated that O(2) generation in the fractions solubilized from pea and cowpea cell walls seems to be catalyzed by cell wall-bound peroxidase(s) and that the plant cell walls alone are able to respond to the elicitor non-specifically and to the suppressor in a species-specific manner, suggesting the plant cell walls may play an important role in determination of plant-fungal pathogen specificity. 相似文献
9.
Kazuhiro Toyoda Eri Yasunaga Masanobu Niwa Yuko Ohwatari Atsushi Nakashima Yoshishige Inagaki Yuki Ichinose Tomonori Shiraishi 《Journal of General Plant Pathology》2012,78(5):311-315
Blue native PAGE analysis for cell wall proteins from pea epicotyls demonstrated that cell wall-associated ecto-apyrase (ATPase) formed a large protein complex(es) ranging from 450 to 900?kDa; one of the components of the complex was copper amine oxidase (CuAO), which catalyzes the oxidation of amines with the subsequent generation of ammonia and hydrogen peroxide. CuAO activity was coordinately regulated in vitro with ATP-hydrolyzing activity by an elicitor and a suppressor from Mycosphaerella pinodes. Moreover, treatment of cell wall proteins with the suppressor caused the appearance of the apyrase monomer. On the basis of these results, M. pinodes may target the apyrase-containing protein complex(es) of the host to attenuate cell wall-based, extracellular defense(s) including the production of hydrogen peroxide. 相似文献
10.
Inducible responses in plants against pathogen attack play a major role in resistance to disease. The defense responses are
mostly associated with the expression of various kinds of inducible genes. We employed differential hybridization to isolate
elicitor-inducible genes (EIGs) of tobacco (Nicotiana tabacum cv. Samsun NN) using the tobacco-fungal elicitor system. A cDNA library was constructed from tobacco leaves treated for 12
hr with hyphal wall components (HWC) prepared from Phytophthora infestans, and six EIGs were identified. Expression of all EIGs was induced after inoculation with the soybean pathogen Pseudomonas syringae pv. glycinea (nonpathogenic on tobacco) or treatment with salicylic acid, and a variety of expression patterns of EIG mRNAs was observed.
Sequence analysis of EIG cDNAs revealed similarities to genes for SAR8.2 (EIG-B39 and EIG-D14), glycine-rich protein (EIG-G7), extensin (EIG-I30), acyltransferase (EIG-I24) and unknown protein (EIG-J7). Possible roles of EIG products in disease resistance are discussed.
Received 30 August 2000/ Accepted in revised form 30 November 2000 相似文献
11.
Chinnapun Dutsadee Churngchow Nunta 《Physiological and Molecular Plant Pathology》2008,72(4-6):179-187
Elicitin and a new protein 75 kDa elicitor were purified from the culture filtrate of Phytophthora palmivora, a pathogen of Hevea brasiliensis (rubber plant). Elicitin was obtained by using a one step of DEAE cellulose chromatography and the new elicitor was obtained by two steps of chromatography: a DEAE cellulose column followed by a hydrophobic column. Both elicitors were stable to heat and a wide range of pH values, but were sensitive to ProteaseK. Both elicitors induced scopoletin, peroxidase isozymes (with substrate o-dianisidine and scopoletin) and total phenolic compounds in cell suspension of H. brasiliensis with similar kinetics. In addition, both elicitors induced peroxidase enzyme (o-dianisidine), total phenolic compounds and enhanced local resistance against P. palmivora on young rubber tree seedlings. However, the increase of peroxidase enzyme and total phenolic compounds in rubber tree seedlings was different from those in cell suspension. Furthermore, during the expression of local resistance the zoospore of P. palmivora induced the peroxidase enzyme (o-dianisidine) more rapidly and with higher level than the control plants. H. brasiliensis is more responsive to the new elicitor than elicitin in triggering defense responses. That is the new elicitor was active at a concentration lower than those required for elicitin, about a 30-fold decrease for activation defense responses in cell suspension. For induction of peroxidase enzyme (o-dianisidine), phenolic compounds and local resistance of rubber plants against P. palmivora, the 75 kDa protein was active at about a 2-fold lower concentration when compared to elicitin. 相似文献
12.
Masashi Amano Kazuhiro Toyoda Akinori Kiba Yoshishige Inagaki Yuki Ichinose Tomonori Shiraishi 《Journal of General Plant Pathology》2013,79(1):12-17
When an elicitor is applied to plants to induce resistance, one of the first detectable events is the efflux of ions from the treated tissue. Here we are the first to demonstrate that an elicitor from Mycosphaerella pinodes evokes leakage of Na+ and K+ ions from isolated cell walls of pea and cowpea in vitro, as observed for epicotyl tissues. Pharmacological experiments showed that this elicitor-stimulated leakage was sensitive to vanadate and N-(3-methylphenyl)biphenyl-4-sulfonamide (NGXT-191), that inhibit a cell wall-associated ATPase (apyrase). Vanadate or NGXT-191 suppressed elicitor-induced superoxide generation and expression of defense genes in vivo. On the basis of these results, we assume that the leakage of these ions, probably associated with an ATP-dependent process(es) in the cell wall, is likely associated with induced defenses of pea and cowpea. 相似文献
13.
Carlos García Mata Lorenzo Lamattina Raúl Oscar Cassia 《European journal of plant pathology / European Foundation for Plant Pathology》2001,107(5):557-562
This work shows that the infection of potato (Solanum tuberosum) detached leaves by the late blight pathogen Phytophthora infestans, was drastically reduced by adding deferoxamine, an exogenous iron chelator. Reactive oxygen species in leaves inoculated with P. infestans were also reduced after adding deferoxamine. A leaf ferritin cDNA fragment was obtained by PCR and used as probe for screening a tuber cDNA library. A cDNA (named StF1) encoding the iron-storing potato ferritin was cloned. StF1 is 915 bp in length and has an open reading frame of 230 amino acids that contains the information for the mature 28 kDa subunit of potato ferritin. StF1 was used as probe in northern blot hybridizations to analyze expression of the ferritin gene. In leaves, ferritin mRNA accumulated in response to pathogen attack. In tubers, ferritin mRNA increased upon treatment with the elicitor eicosapentaenoic acid. These results suggest that iron plays a role in the potato-P. infestans interaction. 相似文献
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15.
为明确帚枝霉属Sarocladium内生生防真菌HND5菌株外泌激发子蛋白SbES的诱导辣椒抗病作用机理,通过构建SbES蛋白的毕赤酵母Pichia pastoris重组蛋白表达菌株,利用纯化后的SbES重组蛋白处理辣椒植株,检测辣椒对棒孢叶斑病的抗性,以及相关抗病反应与抗病基因表达的变化。结果表明,0.1 mg/mL SbES重组蛋白可有效诱导辣椒产生对棒孢叶斑病的抗性,可激发辣椒叶片活性氧爆发、微过敏反应和胼胝质积累等抗病反应;并能有效提高辣椒叶片中与活性氧爆发、过敏性反应、胼胝质合成和植保素合成等抗病反应相关基因,以及水杨酸、茉莉酸和乙烯信号传导关键基因的表达。推测帚枝霉属内生真菌激发子蛋白SbES可通过激活多种抗病信号传导途径来激发辣椒产生对棒孢叶斑病的抗性。 相似文献
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17.
Pseudomonas syringae pv. pisi is a seedborne pathogen distributed worldwide that causes pea bacterial blight. Previous characterization of this pathogen has been carried out with relatively small and/or geographically limited samples. Here, a collection of 91 strains are examined that include strains from recent outbreaks in Spain (53 strains) and from 14 other countries, and that represent all races and the new race 8, including the type race strains. This collection was characterized on the basis of 55 nutritional tests, genetic analysis (rep‐PCR, amplification of AN3 and AN7 specific markers, and multilocus sequence typing (MLST)) and pathogenicity on the differential pea cultivars to identify races. Principal component analysis and distance dendrograms confirm the existence of two genetic lineages within this pathovar, which are clearly discriminated by the AN3/AN7 markers, rep‐PCR and MLST. Strains from races 1 and 7 amplified the AN3 marker; those from races 2, 6 and 8 amplified AN7, while strains of races 3, 4 and 5 amplified either AN3 or AN7. Nevertheless, strains were not grouped by race type by any of the genetic or biochemical tests. Likewise, there was no significant association between metabolic and/or genetic profiling and the geographical origin of the strains. The Spanish collection diversity reflects the variability found in the worldwide collection, suggesting multiple introductions of the bacteria into Spain by contaminated seed lots. 相似文献
18.
B. Schippers Jacoba Sj. Voetberg 《European journal of plant pathology / European Foundation for Plant Pathology》1969,75(4):241-258
Germination of chlamydospores ofFusarium oxysporum f. sp.pisi race 1 in the rhizosphere of pea seedlings and red clover seedlings grown in natural soil heavily infested with the pathogen, was highest in percentage along the actively growing parts of the roots. At these sites, exudation of ninhydrinpositive substances and reducing sugars was most intense with seedlings grown in vitro.No significant difference in the percentage of germinating chlamydospores ofFusarium oxysporum f. sp.pisi race 1 were observed in the rhizosphere soil and on the root surface of homologous parts of roots of seedlings and mature plants of a susceptible Rondo and a resistant Rovar pea cultivar grown in natural soil heavily infested with the pathogen. Differences in the growth of mycelium of the pathogen on the root surface, or in the attachment of the mycelium to the root surface of both cultivars were not observed. Epidermis and cortex cells of roots of both cultivars reacted on penetration by the pathogen by producing a cellulose thickening of the cell wall, which later became infiltrated with a ligning-like material. A selective effect on the activities of the pathogen in the rhizosphere, on the root surface and in the epidermis and cortex in relation to resistance thus could not be demonstrated. Formation of new chlamydospores from germ tubes of germinating chlamydospores was frequently observed in the rhizosphere of the susceptible and resistant pea cultivar and in the rhizosphere of red clover seedlings. 相似文献
19.
Intercellular washing fluid (IWF) obtained from the susceptibleArabidopsis accession Ws-eds1 inoculated withPeronospora parasitica isolate Emoy-2, contained an elicitor of necrosis with ecotype specificity towardsArabidopsis accessions with particular resistance genes. This elicitor caused necrosis on the highly resistant accessions La-er, Nd-1
and partly on Col-5, but not on the susceptible accessions Ws-eds1 and Oy-0. In resistant plants, injection of IWF caused hypersensitive reaction (HR)-like cell collapse which was associated
with the accumulation of phenolics and lignin-like material in walls of cells undergoing cell death. The elicitor is sensitive
to proteinase K and pronase enzymes, heating and autoclaving but insensitive to periodate oxidation, freezing and thawing,
and is not dialyzable. Results suggest that the elicitor is a protein. Fractionation experiments using size-exclusion membranes
revealed that elicitor activity has a molecular weight in excess of 100 kDa.
http://www.phytoparasitica.org posting July 13, 2003. 相似文献
20.
Kana Naito Yasuhiro Ishiga Kazuhiro Toyoda Tomonori Shiraishi Yuki Ichinose 《Journal of General Plant Pathology》2007,73(4):281-285
Flagellin in Pseudomonas syringae is a potent elicitor of defense responses including hypersensitive cell death in dicot plants. The oligopeptides flg22 consisting
of 22 conserved amino acids near the N-terminus of flagellins is reported to induce plant defense responses. Because glycosylation
of the central domain of flagellin affects its elicitor activity, we investigated whether any peptide sequence in addition
to flg22 is required for flagellin-induced hypersensitive reaction. A study of recombinant flagellin polypeptides indicated
that the N-terminal domain including the conserved flg22 is required for flagellin-induced hypersensitive cell death in Arabidopsis thaliana. 相似文献