• Title/Summary/Keyword: chitinase gene A

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

  • 김광호;이대원
    • 한국환경농학회지
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    • 제38권4호
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    • pp.296-306
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    • 2019
  • 온도는 곤충의 발달, 성장, 생식에 중요한 요인이며, 또한 곤충의 생존에 직접적 관련있는 물리적 요인이다. 변온동물인 곤충은 생존을 위해 기후변화에 반응을 해야 하며, 저온과 같은 취약한 환경하에서도 다양한 생존전략을 발달시켜야 한다. 본 연구는 저온에 대한 적응에 기여하는 유전자를 동정하기 위해 배추좀나방 유충의 지방체를 저온과 상온에 노출시켜 전사체 분석을 수행하였다. 저온전사체에서는 chitinase, 표피단백질, Hsp23, chytochrome, Glutathione S transferase, phospholipase 2 유전자의 발현이 증가된 반면, 에너지 대사에 관여하는 UDP-당전이효소, trehalase, trehalose transporter는 오히려 발현이 감소하였다. 저온에 곤충이 노출되었을 때, 대사중심인 지방체의 유전자 발현의 변화가 곤충의 온도 적응을 이해하는 단서를 제공할 수 있을 것으로 기대된다.

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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    • 제38권4호
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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.

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

  • 박성용;방미애;오병준;박정훈;송원섭;최경민;정의수;부희옥;조승식
    • 미생물학회지
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    • 제49권3호
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    • pp.262-269
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    • 2013
  • 본 연구진은 청국장에서 수종의 미생물을 분리하였으며, 이중 소화 효소능이 뛰어난 균주를 선별하였으며, 유전학적 분석결과를 통해 본 균주가 Bacillus 속 균주임을 확인하고 Bacillus subtilis BC-P1이라 명명하였다. Bacillus subtilis BC-P1을 starter로 사용하여 청국장을 제조하였으며, protease, xylanase, chitinase activity, 혈전 분해능, 영양성분 분석 및 아미노산 분석을 통하여 B. subtilis BC-P1를 starter로 활용한 청국장 제품의 개선 가능성을 타진하였다. Bacillus subtilis BC-P1를 starter로 제조한 청국장은 기존 제품에 비해 우수한 소화효소 생성능을 보였으며, 특히 혈전분해효소 생성능이 우수함을 확인하였고, 대조군에 비해 증가된 환원당 및 총 아미노산 함량을 확인 할 수 있었다. 따라서 본 균주를 starter로 하여 청국장을 제조할 경우 기능성분, 생리활성의 증가, 품질 향상을 기대할 수 있을 것으로 판단되며, 향후 대규모 생산시 균주, 제조 조건에 관한 연구가 필요할 것으로 사료된다.

Identification of Differentially Up-regulated Genes in Apple with White Rot Disease

  • Kang, Yeo-Jin;Lee, Young Koung;Kim, In-Jung
    • The Plant Pathology Journal
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    • 제35권5호
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    • pp.530-537
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    • 2019
  • Fuji, a major apple cultivar in Korea, is susceptible to white rot. Apple white rot disease appears on the stem and fruit; the development of which deteriorates fruit quality, resulting in decreases in farmers' income. Thus, it is necessary to characterize molecular markers related to apple white rot resistance. In this study, we screened for differentially expressed genes between uninfected apple fruits and those infected with Botryosphaeria dothidea, the fungal pathogen that causes white rot. Antimicrobial tests suggest that a gene expression involved in the synthesis of the substance inhibiting the growth of B. dothidea in apples was induced by pathogen infection. We identified seven transcripts induced by the infection. The seven transcripts were homologous to genes encoding a flavonoid glucosyltransferase, a metallothionein-like protein, a senescence-induced protein, a chitinase, a wound-induced protein, and proteins of unknown function. These genes have functions related to responses to environmental stresses, including pathogen infections. Our results can be useful for the development of molecular markers for early detection of the disease or for use in breeding white rotresistant cultivars.

유기농업에서 무공해 생물자원을 이용한 병충해 종합방제 기술개발 (I) 키토산의 항균 및 병저항성관련 유전자 유도에 의한 토마토 역병 및 시들음병 억제효과 (Development of Integrated Pest Management Techniques Using Biomass for Organic Farming (I))

  • 오상근;최도일;유승헌
    • 한국식물병리학회지
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    • 제14권3호
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    • pp.278-285
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    • 1998
  • Effects of chitosan on growth of tomato plant, and suppression of Fusaruim wilt caused by Fusarium oxysporum f. sp. lycopersici and late blight casued by Phytophthora infestans, were examined. Both late blight and fusarium wilt were suppressed by spray and irrigation of chitosan, respectively. Inhibition of mycelial growth was not greatly affected by molecular size of chitosan but, concentration dependent effects was observed. Ninty percent of P. infestans and 80% of F. oxysporum f. sp. lycopersici of mycelial growth was inhibited by 1,000 ppm of chitosan (MW 30,000~50,000) when amended in plate media. Induction of defense-related gene expression in plant by chitosan treatments were observed when chitosan treated tobacco and tomato RNA samples were hybridized with several defense-related genes as probes. The results revealed that $\beta$-1,3-glucanase and chitinase genes were strongly induced, while pathogenesis-related protein-1, 3-hydroxy-3-methylglutaryl coenzyme A reductase, anionic peroxidase, phenylalanine ammonia lyase genes were weakly induced by chitosan treatment. These results suggest that chitosan have dual effects on these host-pathogen interactions. Possible roles of chitosan in suppression of tomato diseases by inhibition of mycelial growth and activation of plant defense responses are discussed.

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Characterization of a Phenazine and Hexanoyl Homoserine Lactone Producing Pseudomonas aurantiaca Strain PB-St2, Isolated from Sugarcane Stem

  • Mehnaz, Samina;Baig, Deeba Noreen;Jamil, Farrukh;Weselowski, Brian;Lazarovits, George
    • Journal of Microbiology and Biotechnology
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    • 제19권12호
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    • pp.1688-1694
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    • 2009
  • A novel strain of fluorescent pseudomonad (PB-St2) was isolated from surface-sterilized stems of sugarcane grown in Pakistan. The bacterium was identified as Pseudomonas aurantiaca on the basis of 16S rRNA gene sequence analysis and results from physiological and biochemical characteristics carried out with API50 CH and QTS 24 bacterial identification kits. Assays using substrate-specific media for enzymes revealed lipase and protease activities but cellulase, chitinase, or pectinase were not detected. The bacterium was unable to solubilize phosphate or produce indole acetic acid. However, it did produce HCN, siderophores, and homoserine lactones. In dual culture assays on agar, the bacterium showed antifungal activity against an important pathogen of sugarcane in Pakistan, namely Colletotrichum falcatum, as well as for pathogenic isolates of Fusarium oxysporium and F. lateritium but not against F. solani. The antifungal metabolites were identified using thin-layer chromatography, UV spectra, and MALDI-TOFF spectra and shown to be phenazine-1-carboxylic acid (PCA), 2-hydroxyphenazine (2-OH-PHZ), and N-hexanoyl homoserine lactone (HHL) (assessed using only TLC data). The capacity of this bacterium to produce HCN and 2-OH-PHZ, as well as to inhibit the growth of C. falcatum, has not been previously reported.

Isolation, Characterization, and Use for Plant Growth Promotion Under Salt Stress, of ACC Deaminase-Producing Halotolerant Bacteria Derived from Coastal Soil

  • Siddikee, M.A.;Chauhan, P.S.;Anandham, R.;Han, Gwang-Hyun;Sa, Tong-Min
    • Journal of Microbiology and Biotechnology
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    • 제20권11호
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    • pp.1577-1584
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    • 2010
  • In total, 140 halotolerant bacterial strains were isolated from both the soil of barren fields and the rhizosphere of six naturally growing halophytic plants in the vicinity of the Yellow Sea, near the city of Incheon in the Republic of Korea. All of these strains were characterized for multiple plant growth promoting traits, such as the production of indole acetic acid (IAA), nitrogen fixation, phosphorus (P) and zinc (Zn) solubilization, thiosulfate ($S_2O_3$) oxidation, the production of ammonia ($NH_3$), and the production of extracellular hydrolytic enzymes such as protease, chitinase, pectinase, cellulase, and lipase under in vitro conditions. From the original 140 strains tested, on the basis of the latter tests for plant growth promotional activity, 36 were selected for further examination. These 36 halotolerant bacterial strains were then tested for 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity. Twenty-five of these were found to be positive, and to be exhibiting significantly varying levels of activity. 16S rRNA gene sequencing analyses of the 36 halotolerant strains showed that they belong to 10 different bacterial genera: Bacillus, Brevibacterium, Planococcus, Zhihengliuella, Halomonas, Exiguobacterium, Oceanimonas, Corynebacterium, Arthrobacter, and Micrococcus. Inoculation of the 14 halotolerant bacterial strains to ameliorate salt stress (150 mM NaCl) in canola plants produced an increase in root length of between 5.2% and 47.8%, and dry weight of between 16.2% and 43%, in comparison with the uninoculated positive controls. In particular, three of the bacteria, Brevibacterium epidermidis RS15, Micrococcus yunnanensis RS222, and Bacillus aryabhattai RS341, all showed more than 40% increase in root elongation and dry weight when compared with uninoculated salt-stressed canola seedlings. These results indicate that certain halotolerant bacteria, isolated from coastal soils, have a real potential to enhance plant growth under saline stress, through the reduction of ethylene production via ACC deaminase activity.

Genetic and Phenotypic Diversity of Plant Growth Promoting Rhizobacteria Isolated from Sugarcane Plants Growing in Pakistan

  • Mehnaz, Samina;Baig, Deeba N.;Lazarovits, George
    • Journal of Microbiology and Biotechnology
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    • 제20권12호
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    • pp.1614-1623
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    • 2010
  • Bacteria were isolated from roots of sugarcane varieties grown in the fields of Punjab. They were identified by using API20E/NE bacterial identification kits and from sequences of 16S rRNA and amplicons of the cpn60 gene. The majority of bacteria were found to belong to the genera of Enterobacter, Pseudomonas, and Klebsiella, but members of genera Azospirillum, Rhizobium, Rahnella, Delftia, Caulobacter, Pannonibacter, Xanthomonas, and Stenotrophomonas were also found. The community, however, was dominated by members of the Pseudomonadaceae and Enterobacteriaceae, as representatives of these genera were found in samples from every variety and location examined. All isolates were tested for the presence of five enzymes and seven factors known to be associated with plant growth promotion. Ten isolates showed lipase activity and eight were positive for protease activity. Cellulase, chitinase, and pectinase were not detected in any strain. Nine strains showed nitrogen fixing ability (acetylene reduction assay) and 26 were capable of solubilizing phosphate. In the presence of 100 mg/l tryptophan, all strains except one produced indole acetic acid in the growth medium. All isolates were positive for ACC deaminase activity. Six strains produced homoserine lactones and three produced HCN and hexamate type siderophores. One isolate was capable of inhibiting the growth of 24 pathogenic fungal strains of Colletotrichum, Fusarium, Pythium, and Rhizoctonia spp. In tests of their abilities to grow under a range of temperature, pH, and NaCl concentrations, all isolates grew well on plates with 3% NaCl and most of them grew well at 4 to $41^{\circ}C$ and at pH 11.

Inhibition of Microbial Quorum Sensing Mediated Virulence Factors by Pestalotiopsis sydowiana

  • Parasuraman, Paramanantham;Devadatha, B;Sarma, V. Venkateswara;Ranganathan, Sampathkumar;Ampasala, Dinakara Rao;Reddy, Dhanasekhar;Kumavath, Ranjith;Kim, In-Won;Patel, Sanjay K.S.;Kalia, Vipin Chandra;Lee, Jung-Kul;Siddhardha, Busi
    • Journal of Microbiology and Biotechnology
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    • 제30권4호
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    • pp.571-582
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    • 2020
  • Quorum sensing (QS)-mediated infections cause severe diseases in human beings. The control of infectious diseases by inhibiting QS using antipathogenic drugs is a promising approach as antibiotics are proving inefficient in treating these diseases. Marine fungal (Pestalotiopsis sydowiana PPR) extract was found to possess effective antipathogenic characteristics. The minimum inhibitory concentration (MIC) of the fungal extract against test pathogen Pseudomonas aeruginosa PAO1 was 1,000 ㎍/ml. Sub-MIC concentrations (250 and 500 ㎍/ml) of fungal extract reduced QS-regulated virulence phenotypes such as the production of pyocyanin, chitinase, protease, elastase, and staphylolytic activity in P. aeruginosa PAO1 by 84.15%, 73.15%, 67.37%, 62.37%, and 33.65%, respectively. Moreover, it also reduced the production of exopolysaccharides (74.99%), rhamnolipids (68.01%), and alginate (54.98%), and inhibited the biofilm formation of the bacteria by 90.54%. In silico analysis revealed that the metabolite of P. sydowiana PPR binds to the bacterial QS receptor proteins (LasR and RhlR) similar to their respective natural signaling molecules. Cyclo(-Leu-Pro) (CLP) and 4-Hydroxyphenylacetamide (4-HPA) were identified as potent bioactive compounds among the metabolites of P. sydowiana PPR using in silico approaches. The MIC values of CLP and 4-HPA against P. aeruginosa PAO1 were determined as 250 and 125 ㎍/ml, respectively. All the antivirulence assays were conducted at sub-MIC concentrations of CLP (125 ㎍/ml) and 4-HPA (62.5 ㎍/ml), which resulted in marked reduction in all the investigated virulence factors. This was further supported by gene expression studies. The findings suggest that the metabolites of P. sydowiana PPR can be employed as promising QS inhibitors that target pathogenic bacteria.

Microbiological Features and Bioactivity of a Fermented Manure Product (Preparation 500) Used in Biodynamic Agriculture

  • Giannattasio, Matteo;Vendramin, Elena;Fornasier, Flavio;Alberghini, Sara;Zanardo, Marina;Stellin, Fabio;Concheri, Giuseppe;Stevanato, Piergiorgio;Ertani, Andrea;Nardi, Serenella;Rizzi, Valeria;Piffanelli, Pietro;Spaccini, Riccardo;Mazzei, Pierluigi;Piccolo, Alessandro;Squartini, Andrea
    • Journal of Microbiology and Biotechnology
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    • 제23권5호
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    • pp.644-651
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    • 2013
  • The fermented manure derivative known as Preparation 500 is traditionally used as a field spray in biodynamic agriculture for maintaining and increasing soil fertility. This work aimed at characterizing the product from a microbiological standpoint and at assaying its bioactive properties. The approach involved molecular taxonomical characterization of the culturable microbial community; ARISA fingerprints of the total bacteria and fungal communities; chemical elemental macronutrient analysis via a combustion analyzer; activity assays for six key enzymes; bioassays for bacterial quorum sensing and chitolipooligosaccharide production; and plant hormone-like activity. The material was found to harbor a bacterial community of $2.38{\times}10^8$ CFU/g dw dominated by Gram-positives with minor instances of Actinobacteria and Gammaproteobacteria. ARISA showed a coherence of bacterial assemblages in different preparation lots of the same year in spite of geographic origin. Enzymatic activities showed elevated values of ${\beta}$-glucosidase, alkaline phosphatase, chitinase, and esterase. The preparation had no quorum sensing-detectable signal, and no rhizobial nod gene-inducing properties, but displayed a strong auxin-like effect on plants. Enzymatic analyses indicated a bioactive potential in the fertility and nutrient cycling contexts. The IAA activity and microbial degradation products qualify for a possible activity as soil biostimulants. Quantitative details and possible modes of action are discussed.