• Title/Summary/Keyword: Resistance mechanisms

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Suppression of UDP-glycosyltransferase-coding Arabidopsis thaliana UGT74E2 Gene Expression Leads to Increased Resistance to Psuedomonas syringae pv. tomato DC3000 Infection

  • Park, Hyo-Jun;Kwon, Chang-Seob;Woo, Joo-Yong;Lee, Gil-Je;Kim, Young-Jin;Paek, Kyung-Hee
    • The Plant Pathology Journal
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    • v.27 no.2
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    • pp.170-182
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    • 2011
  • Plants possess multiple resistance mechanisms that protect themselves against pathogen attack. To identify unknown components of the defense machinery in Arabidopsis, gene-expression changes were monitored in Arabidopsis thaliana under 18 different biotic or abiotic conditions using a DNA microarray representing approximately 25% of all Arabidopsis thaliana genes (www.genevestigator.com). Seventeen genes which are early responsive to salicylic acid (SA) treatment as well as pathogen infection were selected and their T-DNA insertion mutants were obtained from SALK institute. To elucidate the role of each gene in defense response, bacterial pathogen Pseudomonas syringae pv. tomato (Pst) DC3000 was inoculated onto individual T-DNA insertion mutants. Four mutants exhibited decreased resistance and five mutants displayed significantly enhanced resistance against Pst DC3000-infection as measured by change in symptom development as compared to wild-type plants. Among them, member of uridin diphosphate (UDP)-glycosyltransferase (UGT) was of particular interest, since a UGT mutant (At1g05680) showed enhanced resistance to Pst-infection in Arabidopsis. In systemic acquired resistance (SAR) assay, this mutant showed enhanced activation of SAR. Also, the enhanced SAR correlated with increased expression of defense-related gene, AtPR1. These results emphasize that the glycosylation of UGT74E2 is a part of the SA-mediated disease-resistance mechanism.

A Comparison between Bacterial Cr(VI) Resistance and Cr(VI) Reduction among Environmental Isolates (미생물의 크롬내성과 크롬환원의 상호 비교)

  • 오영숙;최성찬
    • Korean Journal of Microbiology
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    • v.33 no.2
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    • pp.125-130
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    • 1997
  • Microbial reduction of hexava1ent(VI) to trivalent(lII) chromium is regarded as one of the mechanisms that confers resistance to bacteria. In order to verify this hypothesis, we compared Cr(VI) resistance with Cr(VI) reduction among 20 phenotypically distinct environmental isolates from Cr-contaminated and uncontaminated soils. With glucose as an electron donor, Cr(VI) reduction by washed cell suspensions ranged from 0.014 to 0.305 mM Cr(VI) reduced $h^1$. Cr(VI) resistance of the isolates were measured by growth inhibitions on a liquid medium containing 2 mM Cr(VI) based on their decrease of $A_{630}1$ as compared to the controls without Cr(VI). The isolates had a broad range of resistance from no inhibition to 93.4% inhibition of their growth. Upon correlation analysis, there was no significant relationship between those two phenomena. At a population level, a comparison of % resistant viable counts among the Cr-contaminated and uncontaminated soils showed 19.1 % and 0.4% of their total viable counts, respectively. The difference of % resistance between two site,. strongly suggested that the Cr(VI) present in the soils influences natural selection for resistant phenotypes. However, it is unlikely that the Cr(VI) resistance is dependent solely on the reduction as judged by the correlation analysis.

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Delamanid, Bedaquiline, and Linezolid Minimum Inhibitory Concentration Distributions and Resistance-related Gene Mutations in Multidrug-resistant and Extensively Drug-resistant Tuberculosis in Korea

  • Yang, Jeong Seong;Kim, Kyung Jong;Choi, Hongjo;Lee, Seung Heon
    • Annals of Laboratory Medicine
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    • v.38 no.6
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    • pp.563-568
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    • 2018
  • Background: Delamanid, bedaquiline, and linezolid have recently been approved for the treatment of multidrug- and extensively drug-resistant (MDR and XDR, respectively) tuberculosis (TB). To use these drugs effectively, drug susceptibility tests, including rapid molecular techniques, are required for accurate diagnosis and treatment. Furthermore, mutation analyses are needed to assess the potential for resistance. We evaluated the minimum inhibitory concentrations (MICs) of these three anti-TB drugs for Korean MDR and XDR clinical strains and mutations in genes related to resistance to these drugs. Methods: MICs were determined for delamanid, bedaquiline, and linezolid using a microdilution method. The PCR products of drug resistance-related genes from 420 clinical Mycobacterium tuberculosis strains were sequenced and aligned to those of M. tuberculosis H37Rv. Results: The overall MICs for delamanid, bedaquiline, and linezolid ranged from ${\leq}0.025$ to >1.6 mg/L, ${\leq}0.0312$ to >4 mg/L, and ${\leq}0.125$ to 1 mg/L, respectively. Numerous mutations were found in drug-susceptible and -resistant strains. We did not detect specific mutations associated with resistance to bedaquiline and linezolid. However, the Gly81Ser and Gly81Asp mutations were associated with resistance to delamanid. Conclusions: We determined the MICs of three anti-TB drugs for Korean MDR and XDR strains and identified various mutations in resistance-related genes. Further studies are needed to determine the genetic mechanisms underlying resistance to these drugs.

Pi5 and Pii Paired NLRs Are Functionally Exchangeable and Confer Similar Disease Resistance Specificity

  • Vo, Kieu Thi Xuan;Lee, Sang-Kyu;Halane, Morgan K.;Song, Min-Young;Hoang, Trung Viet;Kim, Chi-Yeol;Park, Sook-Young;Jeon, Junhyun;Kim, Sun Tae;Sohn, Kee Hoon;Jeon, Jong-Seong
    • Molecules and Cells
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    • v.42 no.9
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    • pp.637-645
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    • 2019
  • Effector-triggered immunity (ETI) is an effective layer of plant defense initiated upon recognition of avirulence (Avr) effectors from pathogens by cognate plant disease resistance (R) proteins. In rice, a large number of R genes have been characterized from various cultivars and have greatly contributed to breeding programs to improve resistance against the rice blast pathogen Magnaporthe oryzae. The extreme diversity of R gene repertoires is thought to be a result of co-evolutionary history between rice and its pathogens including M. oryzae. Here we show that Pii is an allele of Pi5 by DNA sequence characterization and complementation analysis. Pii-1 and Pii-2 cDNAs were cloned by reverse transcription polymerase chain reaction from the Pii-carrying cultivar Fujisaka5. The complementation test in susceptible rice cultivar Dongjin demonstrated that the rice blast resistance mediated by Pii, similar to Pi5, requires the presence of two nucleotide-binding leucine-rich repeat genes, Pii-1 and Pii-2. Consistent with our hypothesis that Pi5 and Pii are functionally indistinguishable, the replacement of Pii-1 by Pi5-1 and Pii-2 by Pi5-2, respectively, does not change the level of disease resistance to M. oryzae carrying AVR-Pii. Surprisingly, Exo70F3, required for Pii-mediated resistance, is dispensable for Pi5-mediated resistance. Based on our results, despite similarities observed between Pi5 and Pii, we hypothesize that Pi5 and Pii pairs require partially distinct mechanisms to function.

A Study for the Improvement of Weld Quality Through Force Control of Servo Gun in Resistance Spot Welding using Robot (저항 점 용접 로봇에서 서보건의 가압력 제어를 통한 용접 강도 향상에 대한 연구)

  • Park, Young-Whan;Lee, Jong-Gu;Rhee, Se-Hun
    • Journal of Welding and Joining
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    • v.24 no.6
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    • pp.13-20
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    • 2006
  • Resistance spot welding is widely used for joining sheet metals in the automotive manufacturing process. Recently, servo-gun is used to increase the productivity and precise control the acting force. However, force control mechanisms have not been investigated with servo-guns until now. In this paper, it is proved that servo-motor current is proportional to torque and by experiment, experimental equation between servo-motor current and electrode force was derived. Algorithm for feedback control of electrode force was suggested using current measurement. In addition, applying soft touch method to this system the impact between electrode and specimen, which is the problem of air gun, could be reduced. Indentation made the force decrease in holding time of resistance spot welding. In order to overcome this problem, force compensation using the servo gun was used and it improved weld strength in good welding current range.

Overexpression of cysteine protease in transgenic Brassica rapa enhances resistance to bacterial soft rot and up-regulate the expression of various stress-regulated genes

  • Jung, Yu-Jin;Kang, Kwon-Kyoo
    • Journal of Plant Biotechnology
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    • v.37 no.3
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    • pp.327-336
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    • 2010
  • Cysteine proteases have been known as a critical factor in plant defense mechanisms in pineapple, papaya, or wild fig. Papain or ficin is one kind of cysteine proteases that shows toxic effects to herbivorous insects and pathogenic bacteria. However, resistance to bacterial soft rot of plants genetically engineered with cysteine protease has been little examined thus far. We cloned a cysteine protease cDNA from Ananas comosus and introduced the gene into Chinese cabbage (Brassica rapa) under the control of the cauliflower mosaic virus 35S promoter. The transgene was stably integrated and actively transcribed in transgenic plants. In comparisons with wild-type plants, the $T_2$ and $T_3$ transgenic plants exhibited a significant increase in endo-protease activity in leaves and enhanced resistance to bacterial soft rot. A cDNA microarray analysis revealed that several genes were more abundantly transcribed in the transgenic than in the wild type. These genes encode a glyoxal oxidase, PR-1 protein, PDF1, protein kinase, LTP protein, UBA protein and protease inhibitor. These results suggest an important role for cysteine protease as a signaling regulator in biotic stress signaling pathways, leading to the build-up of defense mechanism to pathogenic bacteria in plants.

Determination of epidemiological tetracycline MIC cut-off value for Vibrio ichthyoenteri

  • Han, Hyun-Ja;Kim, Do-Hyung;Lee, Deok-Chan;Won, Kyoung-Mi;Lee, Soon-Jeong;Cho, Mi-Young;Jee, Bo-Young;Kim, Jin-Woo
    • Journal of fish pathology
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    • v.24 no.2
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    • pp.95-102
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    • 2011
  • Normalized resistance interpretation (NRI) analysis for tetracycline was applied to generate information on the epidemiological cut-off value for Vibrio ichthyoenteri isolated from diseased olive flounder (Paralichthys olivaceus) larvae. Thus, 42 strains of V. ichthyoenteri were used to determine minimum inhibitory concentration (MIC) values of tetracycline using Etest. Also, 11 tetracycline resistance related genes were investigated by PCR method. Most tetracycline-resistant strains harbored both tetB and tetM with a few exceptions. NRI-derived mean and 2 SD above the mean of theoretical normal distributions of susceptible isolates were 0.33 mg/L and 1.66 mg/L, respectively. The epidemiological cut-off value for V. ichthyoenteri from the calculations could be set to S ${\leq}$ 2 mg/L. Of the 42 strains, 15 were classified as non-wild type (NWT), and MIC values of the NWT strains vary regardless of tetB and tetM detection, suggesting that there may be other mechanisms involved in tetracycline resistance in this Vibrio species.

Incidence of Tetracycline Resistance Genes, tet(M) and tet(O), in Streptococci Isolated from Dental Plaques of Koreans

  • Kim, Yeon-Hee;Lee, Si Young
    • International Journal of Oral Biology
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    • v.39 no.1
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    • pp.9-13
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    • 2014
  • Streptococci are among the normal human microflora that populate the oral cavity. However, oral streptococci are known as a major causative agent for dental caries and bacterial endocarditis. Tetracycline is a broad-spectrum antibiotic that is used for oral infections but two mechanisms of tetracycline resistance in streptococci have been reported. The tet(K) and tet(L) genes in these bacteria are related to the active efflux of tetracycline, whereas tet(M) and tet(O) confer ribosomal protection from this antibiotic. It has been reported that the tetracycline resistance of streptococci is related mainly to the activity of tet(M) and tet(O). In our present study, we examined the prevalence of tet(M) and tet(O) in oral streptococci isolated from Korean dental plaques using PCR. One hundred and forty eight of 635 isolates (23.3%) were tetracycline resistant; 68 of these strains (46%) harbored tet(M) and 3 strains (2%) were positive for tet(O). However, tet(M) and tet(O) did not co-exist in any of the resistant strains. Seventy seven of the 148 tetracycline resistant strains (52%) were negative for both the tet(M) and tet(O) genes.

Resistance Mechanism of Acinetobacter spp. Strains Resistant to DW-116, a New Quinolone

  • Choi, Keum-Hwa;Baek, Moon-Chang;Kim, Byong-Kak;Choi, Eung-Chil
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.310-314
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    • 1998
  • DW-116 is a new fluoroquinolone antimicrobial agent with a broad spectrum. In order to elucidate the resistance mechanism to DW-116 in Acinetobacter spp. bacteria, total chromosomal DNA was isolated from 10 strains of Acinetobacter spp. resistant to DW-116. Quinolone resistance determinant region (QRDR) of DNA gyrase gene was amplified by PCR. The 345 bp nucleotide fragment yielded was inserted into pKF 3 which was used as the vector. Comparisons of the DNA sequences of 8 strains with that of the wild type strain revealed a Ser-83 to Leu mutation in mutants and all ten strains contained one silent mutation$(T{\rightarrow}G)$in QRDR. From Acinetobacter MB4-8 strain, DNA gyrase was isolated and purified, through novobiocin-sepharose, heparin-sepharose affinity column chromatography. The enzyme was composed of two subunits and the molecular mass of subunits A and B were 75.6 and 51.9 kDa, respectively. The supercoiling activity of the reconstituted DNA gyrase composed of subunit A from Acinetobacter MB4-8 and subunit B from E. coli was not inhibited by $128{\mu}\textrm{g}$ml of ciprofloxacin. It might be said that one of the resistance mechanisms to DW-116 in Acinetohacter MB4-8 was subunit A alteration of DNA gyrase.

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Analyses of the Less Benzimidazole-sensitivity of the Isolates of Colletotrichum spp. Causing the Anthracnose in Pepper and Strawberry

  • Kim, Yun-Sik;Min, Ji-Young;Kang, Beum-Kwan;Bach, Ngyeun-Van;Choi, Woo-Bong;Park, Eun-Woo;Kim, Heung-Tae
    • The Plant Pathology Journal
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    • v.23 no.3
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    • pp.187-192
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    • 2007
  • The anthracnose disease on pepper fruits in Korea was caused by Colletotrichum acutatum as well as C. gloeosporioides. Since C. acutatum showed less sensitivity to benomyl, it was analyzed whether the less sensitivity was given by the same mechanism for the fungicide resistance of C. gloeosporioides. The isolates of C. acutatum were less sensitive to the three benzimidazole fungicides tested, benomyl, carbendazim, and thiophanate-methyl. However, the of C. acutatum isolates were different from the resistant isolates of C. gloeosporioides in their response to diethofencarb, one of N-phenyl-carbamates; the former was still less sensitive to diethofencarb than the latter. The differences in the resistance mechanisms in two species were conspicuous in sequence analysis of the tub2 genes. The genes from C. acutatum did not show any non-synonymous base substitutions at the regions known to be correlated with the benzimidazole-resistance. All of these data may indicate that the less sensitivity of C. acutatum to benomyl is based on different mechanism(s) from that of C. gloeosporioides.