• Title/Summary/Keyword: Chitinase gene

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Effect of Introducing Chitinase Gene on the Resistance of Tuber Mustard against White Mold

  • Ojaghian, Seyedmohammadreza;Wang, Ling;Xie, Guan-Lin
    • The Plant Pathology Journal
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    • v.36 no.4
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    • pp.378-383
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    • 2020
  • The objective of this research was introduction of chit42 to tuber mustard plants through Agrobacteriummediated transformation against white mold caused by Sclerotinia sclerotiorum. The binary plasmid pGisPEC1 was used in this study. Polymerase chain reaction analysis detected the transgene in 27 transformants with a transformation efficiency of 6.9%. Southern blot test was used to assess the copy number of transgene in tuber mustard plants. One, two, two, and two chit42-related bands were observed in the transformed lines TMB4, TMB7, TMB12, and TMB18, respectively. Enzymatic tests showed a significant increase in the activity of endochitinase in protein isolated from leaf tissues of chit42 transgenic 75-day tuber mustard lines. The pathogenicity of three pathogen isolates was tested on the leaves of transformed plans. The results of current study showed that expression of the gene chit42 in tuber mustard plants markedly reduced infection radius on the leaves 7 days after inoculation with the fungus.

Transciptomic Analysis of Larval Fat Body of Plutella xylostella under Low Temperature (저온조건에서 배추좀나방(Plutella xylostella) 지방체 유전자 발현 변화)

  • Kim, Kwang-Ho;Lee, Dae-Weon
    • Korean Journal of Environmental Agriculture
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    • v.38 no.4
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    • pp.296-306
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    • 2019
  • BACKGROUND: Temperature is known to be the main factor affecting development, growth and reproduction of organisms and also a physical factor directly related to insect survival. Insects as ectothermal species should be responsive to climate changes for their survival and develop various survival strategies under the unfavorable temperature such as low temperature. The purpose of this study is to identify genes contributing to adaptation of low temperature. METHODS AND RESULTS: To identify genes contributing to adaptation of low temperature, the transcriptomic data were obtained from fat body in Plutella xyostella larvae via next generation sequencing. We identified structural proteins, heat shock proteins, antioxidant enzymes, detoxification proteins, and cryoprotectant mobilization and biosynthesis-related proteins. Genes encoding chitinase, cuticular protein, Hsp23, chytochrome protein, Glutathione S transferase, and phospholipase 2 were up-regulated under low temperature. Proteins related to energy metabolism such as UDP-glycosy ltransferase, trehalase and trehalose transporter were down-regulated. CONCLUSION: When insect pests were exposed to low temperature, changes in gene expression of fat body could provide some hints for understanding temperature adaptation strategies.

Characterization of Actinomyces Isolated from Freshwater Sponges in Lake Baikal (바이칼의 담수 스폰지에서 분리한 방선균의 특성 연구)

  • Jung, You-Jung;Joung, Yo-Chan;Ahn, Tae-Seok
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.130-136
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    • 2011
  • Five strains of Actinomyces were isolated from freshwater sponges, Baikalospongia and Lubomirskia, in Lake Baikal. By 16S rRNA sequencing, isolates were identified as Streptomyces griseoplanus, S. halstedii, S. violascens, S. flavovirens, and S. microflavus. Isolates had different characteristics of growth temperature, carbon utilization, enzyme activity, and cellular fatty acid composition. Optimum growth conditions of isolates were $30-37^{\circ}C$, pH 8-9, and 0-1.5% salt concentrations. Major fatty acid compositions were anteiso-$C_{15:0}$, iso-$C_{15:0}$, and iso-$C_{16:0}$. Strain ATS-BA-19 had DNase and chitinase activities and strain ATS-BA-16 had cellulase and protease activities. Colonies of strain ATS-BA-15 and ATS-BA-19 made inhibition zone of Pseudomonas aeruginosa.

Transcriptional Changes of Plant Defense-Related Genes in Response to Clavibacter Infection in Pepper and Tomato

  • Hwang, In Sun;Oh, Eom-Ji;Oh, Chang-Sik
    • The Plant Pathology Journal
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    • v.36 no.5
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    • pp.450-458
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    • 2020
  • Pepper and tomato plants infected with two Clavibacter species, C. capsici and C. michiganensis have shown different patterns of disease development depending on their virulence. Here, we investigated how pepper and tomato plants respond to infection by the high-virulent or low-virulent Clavibacter strains. For this, we chose two strains of each Clavibacter species to show different virulence level in the host plants. Although low-virulent strains showed less disease symptoms, they grew almost the same level as the high-virulent strains in both plants. To further examine the response of host plants to Clavibacter infection, we analyzed the expression patterns of plant defense-related genes in the leaves inoculated with different strains of C. capsici and C. michiganensis. Pepper plants infected with high-virulent C. capsici strain highly induced the expression of CaPR1, CaDEF, CaPR4b, CaPR10, and CaLOX1 at 5 days after inoculation (dai), but their expression was much less in low-virulent Clavibacter infection. Expression of CaSAR8.2 was induced at 2 dai, regardless of virulence level. Expression of GluA, Pin2, and PR2 in tomato plants infected with high-virulent C. michiganensis were much higher at 5 dai, compared with mock or low-virulent strain. Expression of PR1a, Osmotin-like, Chitinase, and Chitinase class 2 was increased, regardless of virulence level. Expression of LoxA gene was not affected by Clavibacter inoculation. These results suggested that Clavibacter infection promotes induction of certain defense-related genes in host plants and that differential expression of those genes by low-virulent Clavibacter infection might be affected by their endophytic lifestyle in plants.

Activation of Pathogenesis-related Genes by the Rhizobacterium, Bacillus sp. JS, Which Induces Systemic Resistance in Tobacco Plants

  • Kim, Ji-Seong;Lee, Jeongeun;Lee, Chan-Hui;Woo, Su Young;Kang, Hoduck;Seo, Sang-Gyu;Kim, Sun-Hyung
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.195-201
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    • 2015
  • Plant growth promoting rhizobacteria (PGPR) are known to confer disease resistance to plants. Bacillus sp. JS demonstrated antifungal activities against five fungal pathogens in in vitro assays. To verify whether the volatiles of Bacillus sp. JS confer disease resistance, tobacco leaves pre-treated with the volatiles were damaged by the fungal pathogen, Rhizoctonia solani and oomycete Phytophthora nicotianae. Pre-treated tobacco leaves had smaller lesion than the control plant leaves. In pathogenesis-related (PR) gene expression analysis, volatiles of Bacillus sp. JS caused the up-regulation of PR-2 encoding ${\beta}$-1,3-glucanase and acidic PR-3 encoding chitinase. Expression of acidic PR-4 encoding chitinase and acidic PR-9 encoding peroxidase increased gradually after exposure of the volatiles to Bacillus sp. JS. Basic PR-14 encoding lipid transfer protein was also increased. However, PR-1 genes, as markers of salicylic acid (SA) induced resistance, were not expressed. These results suggested that the volatiles of Bacillus sp. JS confer disease resistance against fungal and oomycete pathogens through PR genes expression.

Chemical Composition and Quorum Sensing Inhibitory Effect of Nepeta curviflora Methanolic Extract against ESBL Pseudomonas aeruginosa

  • Haitham Qaralleh
    • Journal of Pharmacopuncture
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    • v.26 no.4
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    • pp.307-318
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    • 2023
  • Objectives: Bacterial biofilm is regarded as a significant threat to the production of safe food and the arise of antibiotic-resistant bacteria. The objective of this investigation is to evaluate the quorum sensing inhibitory effect of Nepeta curviflora methanolic extract. Methods: The effectiveness of the leaves at sub-inhibitory concentrations of 2.5, 1.25, and 0.6 mg/mL on the virulence factors and biofilm formation of P. aeruginosa was evaluated. The effect of N. curviflora methanolic extract on the virulence factors of P. aeruginosa, including pyocyanin, rhamnolipid, protease, and chitinase, was evaluated. Other tests including the crystal violet assay, scanning electron microscopy (SEM), swarming motility, aggregation ability, hydrophobicity and exopolysaccharide production were conducted to assess the effect of the extract on the formation of biofilm. Insight into the mode of antiquorum sensing action was evaluated by examining the effect of the extract on the activity of N-Acyl homoserine lactone (AHL) and the expression of pslA and pelA genes. Results: The results showed a significant attenuation in the production of pyocyanin and rhamnolipid and in the activities of protease and chitinase enzymes at 2.5 and 1.25 mg/mL. In addition, N. curviflora methanolic extract significantly inhibited the formation of P. aeruginosa biofilm by decreasing aggregation, hydrophobicity, and swarming motility as well as the production of exopolysaccharide (EPS). A significant reduction in AHL secretion and pslA gene expression was observed, indicating that the extract inhibited quorum sensing by disrupting the quorum-sensing systems. The quorum-sensing inhibitory effect of N. curviflora extract appears to be attributed to the presence of kaempferol, quercetin, salicylic acid, rutin, and rosmarinic acid, as indicated by LCMS analysis. Conclusion: The results of the present study provide insight into the potential of developing anti-quorum sensing agents using the extract and the identified compounds to treat infections resulting from quorum sensing-mediated bacterial pathogenesis.

Fermentation and Quality Characteristics of Cheonggukjang Fermented with Bacillus subtilis BC-P1 (Bacillus subtilis BC-P1 균주를 이용하여 제조한 청국장의 발효 및 품질 특성분석)

  • Park, Sung-Yong;Bang, Mi-Ae;Oh, Boung-Jun;Park, Jeong-Hoon;Song, Won-Seob;Choi, Kyung-Min;Choung, Eui-Su;Boo, Hee-Ock;Cho, Seung-Sik
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.262-269
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    • 2013
  • The object of this study was to improve the quality of Cheonggukjang with new starter, Bacillus subtilis BC-P1. Twenty strains were isolated from the commercial cheonggukjang and 1 Bacillus strain (BC-P1) with protease activity was selected. The 16S rRNA gene sequence revealed that the BC-P1 was closely related to B. subtilis with 99% homology. The quality characteristics of chunggukjang fermented with B. subtilis BC-P1, Bacillus nato (PC) and commercial chunggukjang (NC) were investigated. The characteristics of fermentation were determined by protease, lipase, xylanase, chitinase, and fibrinolytic activities, reducing sugar, nutrient composition and amino acid contents of cheonggukjang sample. Cheonggukjang fermented with B. subtilis BC-P1 showed the strongest fibrinolytic, xylanase, and chitinase activities. Reducung sugar contents of Cheonggukjang samples were $30.16{\pm}2.11$ mg/g (NC), $28.56{\pm}1.52$ mg/g (PC), $32.39{\pm}1.87$ mg/g (BC-P1). And their total amino acid contents were 338.99 mg% (NC), 445.19 mg% (PC), 741.35 mg% (BC-P1). These results suggested that B. subtilis BC-P1 was suitable to be used as a starter to enhance the quality and effects of cheonggukjang.

Identification and Characterization of Genes Differentially Expressed in the Resistance Reaction in Wheat Infected with Tilletia tritici, the Common Bunt Pathogen

  • Lu, Zhen-Xiang;Gaudet, Denis A.;Frick, Michele;Puchalski, Byron;Genswein, Bernie;Laroche, Andre
    • BMB Reports
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    • v.38 no.4
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    • pp.420-431
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    • 2005
  • The differentially virulent race T1 of common bunt (Tilletia tritici) was used to inoculate the wheat lines Neepawa (compatible) and its sib BW553 (incompatible) that are nearly isogenic for the Bt-10 resistance gene. Inoculated crown tissues were used to construct a suppression subtractive hybridization (SSH) cDNA library. Of the 1920 clones arrayed from the SSH cDNA library, approximately 10% were differentially regulated. A total of 168 differentially up-regulated and 25 down-regulated genes were identified and sequenced; 71% sequences had significant homology to genes of known function, of which 59% appeared to have roles in cellular metabolism and development, 24% in abiotic/biotic stress responses, 3% involved in transcription and signal transduction responses. Two putative resistance genes and a transcription factor were identified among the up regulated sequences. The expression of several candidate genes including a lipase, two non-specific lipid transfer proteins (ns-LTPs), and several wheat pathogenesis-related (PR)-proteins, was evaluated following 4 to 32 days post-inoculation in compatible and incompatible interactions. Results confirmed the higher overall expression of these genes in resistant BW553 compared to susceptible Neepawa, and the differential up-regulation of wheat lipase, chitinase and PR-1 proteins in the expression of the incompatible interaction.