• BMB Reports
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• 제39권4호
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• pp.355-360
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• 2006

To gain a better understanding on the function of the potato Solanum tuberosum Multiprotein Bridging Factor 1 protein (StMBF1) its interaction with the TATA box binding protein (TBP) was demonstrated. In addition we reported that StMBF1 rescues the yeast mbf1 mutant phenotype, indicating its role as a plant co-activator. These data reinforce the hypothesis that MBF1 function is also conserved among non closely related plant species. In addition, measurement of StMBF1 protein level by Western blot using anti-StMBF1 antibodies indicated that the protein level increased upon $H_2O_2$ and heat shock treatments. However, the potato $\beta$-1,3-glucanase protein level was not changed under the same experimental conditions. These data indicate that StMBF1 participates in the cell stress response against oxidative stress allowing us to suggest that MBF1 genes from different plant groups may share similar functions.

• Journal of the Korean Wood Science and Technology
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• 제46권6호
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• pp.670-680
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• 2018

Acanthophysium sp. KMF001, a wood rotting fungus, produces a strong crude enzyme complex that efficiently produces simple sugars from wood. The transcriptomic analysis of Acanthophysium sp. KMF001 identified 14 genes for putative glycoside hydrolases. Among them, isotig01043 was expressed heterogeneously in Escherichia coli BL21(DE3), and the expressed protein exhibited an endo-${\beta}$-1,4-xylanase activity which showed the optimum reaction at pH 5.0 and $30^{\circ}C$. The enzyme kinetic values of $K_m$ and $V_{max}$ were 25.92 mg/ml and $0.628{\mu}mole/mg/ml$, respectively. The enzymatic characteristics of the expressed xylanase showed a typical fungal xylanase. However, the bioinformatics analysis suggested that the protein encoded by isotig01043 was a novel xylanase based on a low identity when it was compared with the closest protein in the NCBI database and a similar protein domain with GH16_fungal_Lam16A_glucanase, which had not been earlier suggested as a xylanase.

• Journal of Microbiology and Biotechnology
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• 제22권8호
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• pp.1118-1126
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• 2012

Four putative GH12 genes were found in the Fusarium graminearum genome. The corresponding proteins were expressed in Escherichia coli, purified, and evaluated. FGSG_05851 and FGSG_11037 displayed high activities towards xyloglucan ($V_{max}$ of 4 and $11{\mu}mol/min$, respectively), whereas FGSG_07892 and FGSG_16349 were much less active with this substrate (0.081 and $0.004{\mu}mol/min$, respectively). However, all four of these enzymes had a similar binding affinity for xyloglucan. Xyloglucan was the substrate preferred by FGSG_05851, in contrast to the three other enzymes, which preferred ${\beta}$-glucan or lichenan. Therefore, FGSG_05851 is a xyloglucan-specific glucanase (E.C. 3.2.1.151) rather than an endoglucanase (E.C. 3.2.1.4) with broad substrate specificity. FGSG_11037 displayed a peculiar behavior in that the xyloglucan binding was highly cooperative, with a Hill coefficient of 2.5. Finally, FGSG_05851 essentially degraded xyloglucan into hepta-, octa-, and nonasaccharides, whereas the three other enzymes yielded hepta- and octa-saccharides as well as larger molecules.

• Journal of Microbiology and Biotechnology
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• 제24권10호
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• pp.1327-1336
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• 2014

Bacillus amyloliquefaciens strain NJZJSB3 has shown antagonism of several phytopathogens in vitro, especially Sclerotinia sclerotiorum. Both the broth culture and cell suspension of strain NJZJSB3 could completely protect the detached leaves of canola (Brassica napus) from S. sclerotiorum infection. In pot experiments, the application of strain NJZJSB3 cell suspension ($10^8CFU/ml$) decreased the disease incidence by 83.3%, a result similar to commercially available fungicide (Dimetachlone). In order to investigate the potential biocontrol mechanisms of strain NJZJSB3, the nonvolatile antifungal compounds it produces were identified as iturin homologs using HPLC-ESI-MS. Antifungal volatile organic compounds were identified by gas chromatography-mass spectrometry. The detected volatiles toluene, phenol, and benzothiazole showed antifungal effects against S. sclerotiorum in chemical control experiments. Strain NJZJSB3 also produced biofilm, siderophores and cell-wall-degrading enzymes (protease and ${\beta}$-1,3-glucanase). These results suggest that strain NJZJSB3 can be a tremendous potential agent for the biological control of sclerotinia stem rot.

• 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.

• BMB Reports
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• 제34권4호
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• pp.310-315
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• 2001

The uneven distribution of acidic and basic chitinases in different parts of rice seed, and also the characterization of hull-specific chitinases, are reported here. After extraction of chitinases from polished rice, bran, and rice hulls, the chitinases were separated into acidic and basic fractions, according to their behavior on an anion exchanger column. Both fractions from different parts of rice seed showed characteristic activity bands on SDS-PAGE that contained 0.01% glycol chitin. The basic chitinases from rice hulls were further purified using chitin affinity chromatography. The chitinase, specific to rice hulls (RHBC), was 88-fold purified with a 1.3% yield. RHBC has an apparent molecular weight of 22.2 kDa on SDS-PAGE. The optimal pH and temperature were 4.0 and $35^{\circ}C$, respectively. With [$^3H$]chitin as a substrate, RHBC has $V_{max}$ of 13.51 mg/mg protein/hr and $K_m$ of 1.36 mg/ml. This enzyme was an endochitinase devoid of ${\beta}$-1,3-glucanase, lysozyme, and chitosanase activities.

• Journal of Plant Biotechnology
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• 제43권1호
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• pp.37-48
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• 2016

The transcriptional profiles of 'Tamnara' grapevine (Vitis labruscana L.) to Rhizobium vitis were determined using 12,000 gene oligonucleotide microarray chips constructed with 6,776 unigenes based on the EST sequencing. Among them, 95 clones were up-regulated more than three times and 90 were down-regulated more than 5-times in the R. vitis-inoculated grapevines relative to the control vines. Treatment of salicylic acid showed that 337 clones were upregulated and 52 clones were down regulated in grapevines. Microarray analysis, reverse transcription-polymer chain reaction, and slot blot hybridization analysis revealed that 5, 14, and 64 clones were up-regulated and 10, 12, and 61 clones were down-regulated in wounded, salicylic acid-treated, and R. vitis-inoculated 'Tamnara' grapevine leaves, respectively. The expression patterns of ${\beta}$-1,3-glucanase, proline-rich protein, and lipoxygenase genes of 'Tamnara' moderately resistant to R. vitis were similar to those of resistant 'Concord' and 'Delaware' grapevines. However, chalcone synthase genes in 'Tamnara' grapevines showed similar expression patterns to susceptible grapevines 'Neomuscat' and 'Rizamat'. Further expression studies with various clones for each gene should be conducted to elucidate their roles in resistant responses against pathogens or other stimuli in grapevines. These results could provide better resources for understanding the mechanism of defense responses against crown gall disease and clues for identifying new genes that may play a role in defense against R. vitis in grapevines.

• The Plant Pathology Journal
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• 제29권1호
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• pp.77-86
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• 2013

Anthracnose development by Colletotrichum musae was observed to be significantly less in the fruits of the banana cultivar 'Embul' (Mysore, AAB) infected with Phyllosticta musarum than in fruits without such infections. Anthracnose disease originates from quiescent C. musae infections in the immature fruit. P. musarum incites minute, scattered spots, referred to as freckles, in the superficial tissues of immature banana peel which do not expand during maturation or ripening. P. musarum does not appear to have a direct suppressive effect on C. musae as conidia of C. musae germinate on both freckled and non-freckled fruit forming quiescent infections. Our investigations have shown that P. musarum infection induced several defence responses in fruit including the accumulation of five phytoalexins, upregulation of chitinase and ${\beta}$-1,3-glucanase, phenylalanine ammonia lyase (PAL) activity and cell wall lignification. $^1H$ and $^{13}C$ NMR spectral data of one purified phytoalexin compared closely with 4'-hydroxyanigorufone. Some of the P. musarum-induced defences that retained during ripening, restrict C. musae development at the ripe stage. This paper examines the potential of P. musarum-induced defences, in the control of anthracnose, the most destructive postharvest disease in banana.

• 한국해양바이오학회지
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• 제2권2호
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• pp.108-112
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• 2007

A cellulase (endo-${\beta}$-1,4-D-glucanase(E.C.3.2.1.4)) was isolated from the hepatopancreas of abalone Haliotis discus hannai by EST analysis. The abalone cellulase named HdEG compassed 1977 bp, including 195 bp in the 5'untranslated region, 1680 bp in the open reading frame which encodes 560 amino acid residues, and 92 bp in the 3'-untranslated region. The C-terminal region of the HdEG showed 44-52% identity to the catalytic domains of glycoside hydrolase family 9 (GHF9)-cellulases from arthropods and bacteria. The recombinant cellulase, pEHdEG was produced in E. coli with being fused with C-terminal His-tag. The expressed protein showed a single band (~62 kDa) on Western blotting which was consistent with the value (61,878 Da) calculated from the DNA sequence.

• Mycobiology
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• 제43권3호
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• pp.311-318
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• 2015

Culture filtrates of six different edible mushroom species were screened for antimicrobial activity against tomato wilt bacteria Ralstonia solanacearum B3. Hericium erinaceus, Lentinula edodes (Sanjo 701), Grifola frondosa, and Hypsizygus marmoreus showed antibacterial activity against the bacteria. Water, n-butanol, and ethyl acetate extracts of spent mushroom substrate (SMS) of H. erinaceus exhibited high antibacterial activity against different phytopathogenic bacteria: Pectobacterium carotovorum subsp. carotovorum, Agrobacterium tumefaciens, R. solanacearum, Xanthomonas oryzae pv. oryzae, X. campestris pv. campestris, X. axonopodis pv. vesicatoria, X. axonopodis pv. citiri, and X. axonopodis pv. glycine. Quantitative real-time PCR revealed that water extracts of SMS (WESMS) of H. erinaceus induced expressions of plant defense genes encoding ${\beta}$-1,3-glucanase (GluA) and pathogenesis-related protein-1a (PR-1a), associated with systemic acquired resistance. Furthermore, WESMS also suppressed tomato wilt disease caused by R. solanacearum by 85% in seedlings and promoted growth (height, leaf number, and fresh weight of the root and shoot) of tomato plants. These findings suggest the WESMS of H. erinaceus has the potential to suppress bacterial wilt disease of tomato through multiple effects including antibacterial activity, plant growth promotion, and defense gene induction.