• Title/Summary/Keyword: Nonpathogenic mutant

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Identification of hrcC, hrpF, and maA Genes of Xanthomonas campestris pv. glycines 8ra: Roles in Pathogenicity and Inducing Hypersensitive Response on Nonhost Plants

  • Park, Byoung-Keun;Ingyu Hwang
    • The Plant Pathology Journal
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    • v.15 no.1
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    • pp.21-27
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    • 1999
  • Nonpathogenic mutants of Xanthomonas campestris pv. glycines were generated with Omegon-Kim to isolate genes essential for pathogenicity and inducing hypersensitive response (HR). Three nonpathogenic multants and two mutants showing slow symptom development were isolated among 1,000 colonies tested. From two nonpathogenic mutants, 8-13 and 26-13, genes homologous to hrcC and hrpF of X. campestris pv. vesicatoria were identified. The nonpathogenic mutant 8-13 had a mutation in a gene homologous to hrpF of X. campestris pv. vesicatoria and failed to cause HR on pepper plants but still induced HR on tomato leaves. The nonpathogenic mutant 26-13 had an insertional mutation in a gene homologous to hrcC of X. campestris pv. vesicatoria and lost the ability to induce HR on pepper leaves but still caused HR on tomato plants. Unlike other phytopathogenic bacteria, the parent strain and these two mutants of X. campestris pv. glycines did not cause HR on tobacco plants. a cosmid clone, pBL1, that complemented the phenotypes of 8-13 was isolated. From the analysis of restriction enzyme mapping and deletion analyses of pBL1, a 9.0-kb Eco RI fragment restored the phenotypes of 8-13. pBL1 failed to complement the phenotypes of 26-13, indicating that the hrcC gene resides outside of the insert DNA of pBL1. One nonpathogenic mutant, 13-33, had a mutation in a gene homologous to a miaA gene encoding tRNA delta (2)-isopentenylpyrophosphate transferase of Escherichia coli. This indicated that tRNA modifications in X. campestris pv. glycines may be required for expression of genes necessary for pathogenicity. The mutant 13-33 multiplied as well as the parent strain did in the culture medium and in planta, indicating that loss of pathogenicity is not due to the inability of multiplication in vivo.

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Characterization of Nonhypersensitive Mutant and Nonpathogenic Mutant of Xanthomonas campestris pv. campestris (Xanthomonas campestris pv. campestris의 비과민성 돌연변이주와 비병원성 돌연변이주의 특성)

  • Kim, Mee-Hyang;Bae, Dong-Won;Lee, Jun-Teak;Yun, Han-Dae;Kim, Hee-Kyu
    • Korean Journal Plant Pathology
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    • v.13 no.3
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    • pp.179-183
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    • 1997
  • We have screened hypersensitive responses of 18 cultivars of Nicotiana tabacum to Xanthomonas campestris pv. campestris. TC500 cultivar produced the most strong hypersensitive response (HR) to Xanthomonas campestris pv. campestris. By NTG mutagenesis, nonhypersensitive mutants (XHN 514-774, XHN 620-831) were generated, which does not induce hypersensitive response on tobacco leaves (Nicotiana tabacum cv. TC500). Also nonpathogenic mutant (XPN 1001), which does not incite any of the black rot symptoms on leaves was generated. We observed that HR mutants were still pathogenic on cabbage leaves producing black rot symptoms and nonpathogenic mutant induced HR in tobacco leaves. The inplanta growth of wild type and HR mutants were examined for up to 120 hrs after inoculation : population of wild type strain increased to $10^{8}$ in 24hrs, but rapidly declined thereafter; HR-mutants increased to more than $10^{6}$ in 48 hrs after inoculation but subsequently stabilized and slowly decreased. We observed that wild type and these mutans produced similar amounts of degradative enzymes such as protease, pectate lyase, cellulase and amylase.

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Biological Control of Fusarium Wilt of Spinach by Nonpathogenic Isolates of Fusarium oxysporum (비병원성 Fusarium oxysporum 구조를 이용한 시금치 시들음병의 생물학적 방제)

  • 신동범;죽원이명
    • Korean Journal Plant Pathology
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    • v.14 no.2
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    • pp.145-149
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    • 1998
  • Four nonpathogenic isolates of Fusarium oxysporum isolated from spinach showed suppressive effect on the occurrence of the Fusarium wilt of spinach caused by F. oxysporum f. sp. sprinaciae, among which NF01 controlled the disease most effectively. And NF01 was not pathogenic to tomato, cucumber, radish and spinach. This isolate was further tested for the biological control of the disease. The isolate was not inhibitory to the growth of the pathogen on potato sucrose agar medium, however the Fusarium wilt disease occurred less by drenching spore suspension of the nonpathogenic isolate. The control effect of the isolate was higher at lower inoculum level of the pathogen than at the higher inoculum level, and in the pretreatments than the simultaneous treatment of the isolate with the pathogen inoculation. The nit mutants of the isolate were easily formed on chlorate containing media, and was reisolated selectively as nit mutant from infected soil and plants. The reisolation rate of the isolate as opposed to pathogen was high at preinoculated soil and plants relative to the simultaneous inoculation of the isolate with the pathogen.

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Suppression Effect on Soft-rot by Bacteriocin-producing Avirulent Pectobacterium carotovorum subsp. carotovorum Pcc21-M15 (박테리오신을 분비하는 비병원성 돌연변이주에 의한 무름병 방제 효과)

  • Roh, Eun-Jung;Lee, Seung-Don;Heu, Sung-Gi
    • Research in Plant Disease
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    • v.16 no.2
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    • pp.136-140
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    • 2010
  • Pectobacterium carotovorum subsp. carotovorum causes soft rot disease in diverse plants. Carocin D is bacteriocin that is produced by Pectobacterium carotovorum subsp. carotovorum Pcc21 strain. Nonpathogenic mutant P. carotovorum subsp. carotovorum Pcc21-M15 strain was obtained by mutagenesis with Tn5 insertion and screened pathogenesity. P. carotovorum subsp. carotovorum Pcc21-M15 and E. coli (pRG3431), carocin D gene-transformed E. coli, produce carocin D against P. carotovorum subsp. carotovorum Pcc3. Pathogenic P. carotovorum subsp. carotovorum Pcc3 and mixture with Pcc21-M15 or E. coli (pRG3431) were treated with lettuces. Pcc21-M15 and E. coli (pRG3431) effectively suppressed the development of soft-rot disease. While symptoms in 90% of Pcc3-treated lettuces were observed after 3 days, only 25% of Pcc3 and Pcc21-M15-treated lettuces were observed to be infected after 6 days. These results suggest that the nonpathogenic strain P. carotovorum subsp. carotovorum. Pcc21-M15 and E. coli (pRG3431) are effective to soft-rot disease suppression.

Effect of Scutellariae Radix as a Novel Antibacterial Herb on the ppk(Polyphosphate Kinase) Mutant of Salmonella typhimurium

  • Hahm, Dae-Hyun;Yeom, Mi-Jung;H.Lee, Eun-Joo;Shim, In-Sop;Lee, Hye-Jung;Kim, Hong-Yeoul
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1061-1065
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    • 2001
  • The antibacterial effects of water extracts of Scutellariate Radix (a dried root of Scutellaria baicalensis GEORGI) and its major flavonoid components, Baicalin and Baicalein, on Salmonella typhimurium, a representative enteric pathogen, were studied. Through a Kriby-Bauer disc analysis, the growth-inhibition activity of Scutellariae Radix against. S. typhimurium was found to be compatible with commercial antibiotics, such as ampicillin, chloramphenicol, and streptomycin. In contrast, the growth of a nonpathogenic E. coli strain was unaffercted by Scutellariae Radix. To examine the effect of polyphosphate kinase (ppk), a putative virulence factor, on the antibacterial activity of Scutellariae Radix, the growth profile of a ppk mutant of S. typhimurium was investigated in a tryptic soy broth containing different concentrations of water extracts of Scutellariae Radix. The ppk mutant was able to grow in 6 mg/ml of water extracts of Scutellariae Radix, whereas in 6 mg/ml of water extracts of Scutellariae Radix, whereas the wild-type could not, implying that the inactivation of ppk made S. typhimurium more resistant to the antibacterial activity of Scutellariae Radix. No enhanced resistance was observed in a ppk mutant of S. typhimurium complemented with a ppk expression vector. The attenuation of the virulence by ppk inactivation was also observed in a virulence assay using BLAB/c mice. Neither Baicalin nor Baicalein exhibited any growth-inhibition activity against S. typhimurium. The water extracts of Scutellariae Radix stimulated the transcription of ppk, especially in the early growth-stage of S. typhimurium.

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A Short-chain Dehydrogenase/reductase Gene is Required for Infection-related Development and Pathogenicity in Magnaporthe oryzae

  • Kwon, Min-Jung;Kim, Kyoung-Su;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • v.26 no.1
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    • pp.8-16
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    • 2010
  • The phytopathogenic fungus Magnaporthe oryzae is a major limiting factor in rice production. To understand the genetic basis of M. oryzae pathogenic development, we previously analyzed a library of T-DNA insertional mutants of M. oryzae, and identified ATMT0879A1 as one of the pathogenicity-defective mutants. Molecular analyses and database searches revealed that a single TDNA insertion in ATMT0879A1 resulted in functional interference with an annotated gene, MGG00056, which encodes a short-chain dehydrogenase/reductase (SDR). The mutant and annotated gene were designated as $MoSDR1^{T-DNA}$ and MoSDR1, respectively. Like other SDR family members, MoSDR1 possesses both a cofactor-binding motif and a catalytic site. The expression pattern of MoSDR1 suggests that the gene is associated with pathogenicity and plays an important role in M. oryzae development. To understand the roles of MoSDR1, the deletion mutant ${\Delta}Mosdr1$ for the gene was obtained via homology-dependent gene replacement. As expected, ${\Delta}Mosdr1$ was nonpathogenic; moreover, the mutant displayed pleiotropic defects in conidiation, conidial germination, appressorium formation, penetration, and growth inside host tissues. These results suggest that MoSDR1 functions as a key metabolic enzyme in the regulation of development and pathogenicity in M. oryzae.