• 한국미생물·생명공학회지
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• 제51권4호
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• pp.449-456
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• 2023

Sugar or dextrose increases the cost of production of xanthan gum by Xanthomonas campestris. Brewers' Spent Grain (BSG) was chosen as a source of fermentable sugars. BSG is a significant industrial by-product generated in large quantities from the breweries. Primarily used as animal feed due to its high fiber and protein content, BSG holds great potential as an economically and ecologically sustainable substrate for fermenting biomolecules. This study explores BSG's potential as a cost-effective carbon source for producing xanthan, utilizing Xanthomonas campestris NCIM 2961. An aqueous extract was prepared from BSG and inoculated with the bacterium under standard fermentation conditions. After fermentation, xanthan gum was purified using a standard protocol. The xanthan yield from BSG media was compared to that from MGYP media (control). The fermentation parameters, including pH, temperature, agitation and duration were optimized for maximum xanthan gum yield by varying them at different levels. Following fermentation, the xanthan gum was purified from the broth by alcoholic precipitation and then dried. The weight of the dried gum was measured. The obtained xanthan from BSG under standard conditions and commercial food-grade xanthan were characterized using FTIR. The highest xanthan yields were achieved at 32 ℃, pH 6.0, and 72 h of fermentation at 200 rpm using BSG media. The FTIR spectra of xanthan from BSG media closely resembled that of commercial food-grade xanthan. The results confirm the potential of BSG as a cost-effective alternative carbon source for xanthan production, thereby reducing production costs and solid waste.

• The Plant Pathology Journal
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• 제31권3호
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• pp.212-218
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• 2015

In this study, we developed a species-specific PCR assay for rapid and accurate detection of three Xanthomonas species, X. axonopodis pv. poinsettiicola (XAP), X. hyacinthi (XH) and X. campestris pv. zantedeschiae (XCZ), based on their draft genome sequences. XAP, XH and XCZ genomes consist of single chromosomes that contain 5,221, 4,395 and 7,986 protein coding genes, respectively. Species-specific primers were designed from variable regions of the draft genome sequence data and assessed by a PCR-based detection method. These primers were also tested for specificity against 17 allied Xanthomonas species as well as against the host DNA and the microbial community of the host surface. Three primer sets were found to be very specific and no amplification product was obtained with the host DNA and the microbial community of the host surface. In addition, a detection limit of $1pg/{\mu}l$ per PCR reaction was detected when these primer sets were used to amplify corresponding bacterial DNAs. Therefore, these primer sets and the developed species-specific PCR assay represent a valuable, sensitive, and rapid diagnostic tool that can be used to detect three specific pathogens at early stages of infection and may help control diseases.

• The Plant Pathology Journal
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• 제32권6호
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• pp.500-507
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• 2016

Single-molecule real-time (SMRT) sequencing allows identification of methylated DNA bases and methylation patterns/motifs at the genome level. Using SMRT sequencing, diverse bacterial methylomes including those of Helicobacter pylori, Lactobacillus spp., and Escherichia coli have been determined, and previously unreported DNA methylation motifs have been identified. However, the methylomes of Xanthomonas species, which belong to the most important plant pathogenic bacterial genus, have not been documented. Here, we report the methylomes of Xanthomonas axonopodis pv. glycines (Xag) strain 8ra and X. campestris pv. vesicatoria (Xcv) strain 85-10. We identified $N^6$-methyladenine (6mA) and $N^4$-methylcytosine (4mC) modification in both genomes. In addition, we assigned putative DNA methylation motifs including previously unreported methylation motifs via REBASE and MotifMaker, and compared methylation patterns in both species. Although Xag and Xcv belong to the same genus, their methylation patterns were dramatically different. The number of 4mC DNA bases in Xag (66,682) was significantly higher (29 fold) than in Xcv (2,321). In contrast, the number of 6mA DNA bases (4,147) in Xag was comparable to the number in Xcv (5,491). Strikingly, there were no common or shared motifs in the 10 most frequently methylated motifs of both strains, indicating they possess unique species- or strain-specific methylation motifs. Among the 20 most frequent motifs from both strains, for 9 motifs at least 1% of the methylated bases were located in putative promoter regions. Methylome analysis by SMRT sequencing technology is the first step toward understanding the biology and functions of DNA methylation in this genus.

• Journal of Microbiology and Biotechnology
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• 제23권1호
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• pp.22-28
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• 2013

Xo2276 is a putative transcription activator-like effector (TALE) in Xanthomonas oryzae pv. oryzae (Xoo). Xo2276 was expressed with a TAP-tag at the C-terminus in Xoo cells to enable quantitative analysis of protein expression and secretion. Nearly all TAP-tagged Xo2276 existed in an insoluble form; addition of rice leaf extracts from a Xoosusceptible rice cultivar, Milyang23, significantly stimulated secretion of TAP-tagged Xo2276 into the medium. In a T3SS-defective Xoo mutant strain, secretion of TAPtagged Xo2276 was blocked. Xo2276 is a Xoo ortholog of Xanthomonas campestris pv. vesicatoria (Xcv) AvrBs3 and contains a conserved DNA-binding domain (DBD), which includes 19.5 tandem repeats of 34 amino acids. Xo2276- DBD was expressed in E. coli and purified. Direct in vitro recognition of Xo2276-DBD on a putative target DNA sequence was confirmed using an electrophoretic mobility shift assay. This is the first study measuring the homologous expression and secretion of Xo2276 in vitro using rice leaf extract and its direct in vitro binding to the specific target DNA sequence.

• The Plant Pathology Journal
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• 제18권2호
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• pp.63-67
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• 2002

To understand plant-pathogen interactions, a complete set of hot pepper genes differentially expressed against pathogen attack was isolated. As an initial step, hundreds of differentially expressed cDNAS were isolated from hot pepper leaves showing non-host resistance against bacterial plant pathogens (Xanthomonas campestris pv. glycines and Pseudomonas syringae pv. syringae) using differential display reverse transcription polymerase chain reaction (DDDRT-PCR) technique. Reverse Northern and Northern blot analyses revealed that 50% of those genes were differentially expressed in pepper loaves during non-host resistance response. Among them, independent genes without redundancy were micro-arrayed for further analysis. Random EST sequence database were also generated from various CDNA libraries including pepper tissue specific libraries and leaves showing non-host hypersensitive response against X. campestris pv. glycines. As a primary stage, thousands of cDNA clones were sequenced and EST data were analyzed. These clones are being spotted on glass slide to study the expression profiling. Results of this study may further broaden knowledge on plant-pathogen interactions.

• The Plant Pathology Journal
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• 제36권2호
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• pp.157-169
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• 2020

Two lactic acid bacteria (LAB) designated J02 and J13 were recovered from fermented vegetables based on their ability to suppress soft rot disease caused by Pectobacterium carotovorum subsp. carotovorum (Pcc) on radish. J02 and J13 were identified as Lactobacillus pentosus and Leuconostoc fallax, respectively. The ability of J02 and J13 to suppress plant diseases is highly dependent on chitosan. LAB alone has no effect and chitosan alone has only a moderate effect on disease reduction. However, J02 or J13 broth cultures plus chitosan display a strong inhibitory effect against plant pathogens and significantly reduces disease severity. LAB strains after being cultured in fish surimi (agricultural waste) and glycerol or sucrose-containing medium and mixed with chitosan, reduce three cruciferous vegetable diseases, including cabbage black spot caused by Alternaria brassicicola, black rot caused by Xanthomonas campestris pv. campestris, and soft rot caused by Pcc. Experimental trials reveal that multiple applications are more effective than a single application. In-vitro assays also reveal the J02/chitosan mixture is antagonistic against Colletotrichum higginsianum, Sclerotium rolfsii, and Fusarium oxysporum f. sp. rapae, indicating a broad-spectrum activity of LAB/chitosan. Overall, our results indicate that a synergistic combination of LAB and chitosan offers a promising approach to biocontrol.

• 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.

• 미생물학회지
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• 제40권3호
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• pp.183-188
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• 2004

TNT 분해 세균 Stenotrophomonas sp. OK-5는 세 개의 다른 NAD(P)H-nitroreductase의 활성 fractions (NTR fractions I, II, III)을 갖고 있는 것으로 확인된 바 있다. 본 연구에서는 NTR fractions I, II, III에 대한 생리학적 특성과 분자생물학적 특성을 규명하고자 하였다. TNT에 대한 균주 OK-5의 NTR fractions I, II, 그리고 III의 활성은 억제 물질인 $\beta$-mercaptoethanol의 첨가 시에 효소의 활성 이 모두 억제되는 것으로 확인되었다. TNT와 그 유사 기질을 이용하여 균주 OK-5에서 분리된 NTR의 기질 특이성을 조사한 결과, nitrobenzene, 그리고 RDX에 대 해서는 비교적 활성 이 높게 나타났으나 2,6-DNT와 2,4-DNT에서는 낮은 활성을 나타내는 것으로 확인되었다. 균주 OK-5에서 정제된 NTR fraction I의 N-말단 아미노산 서 열은 $^1MSDLLNADAVVQLFRTARDS^20$로 분석되었고, Xanthomonas campestris의 NTR과 X. axonopodis의 NTR에서 각각 70%와 65%로 비교적 높은 유사성을 가지는 것으로 나타났다. 균주 OK-5의 NTR fraction I의 효소를 암호화하는 SmOK5nrI 유전자의 염기서열을 확인하고 분석된 유전자로부터 유추되는 아미노산 서열을 각각 비교한 결과 X. campestris의 NTR과 81%, X. axonopodis의 NTR과 75%,그리고 Streptomyces avermitilis의 NTR과 30%의 유사성이 있는 것으로 조사되었으나, Pseudomonas putida KT2440의 NTR (pnrB)과는 16%로 낮은 유사성이 있는 것으로 확인되었다.

• 한국응용곤충학회지
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• 제24권2호
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• pp.97-101
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• 1985

한국(韓國)에서 발생(發生)하고 있는 흰잎마름병균의 병원성(病原性) 변이연구(變異硏究)는 일본판별품종(日本判別品種)에 의한 일본(日本) 분류체계(分類體系)를 이용(利用)하여 전국적(全國的)인 균형(菌型)의 분포상태(分布狀態)를 조사(調査)하여 왔으나 일본판별품종(日本判別品種)을 사용(使用)한 균형분류(菌型分類) 결과(結果) 동일(同一) 균군중(菌群中)에서도 다수계품종(多收系品種)에 따라 상이(相異)한 반응(反應)이 나타나 일본판별품종(日本判別品種)을 이용(利用)한 균형분류(菌型分類)가 부적합(不適合)한 것으로 나타났고, 일본판별품종(日本判別品種)을 한국(韓國)에서 증식(增殖) 보존(保存)할때 출수(出穗)가 늦거나 도복(倒伏)에 약(弱)하여 채종(採種)하기 어려운 점이 있으므로 한국(韓國)에 알맞는 판별품종(判別品種)을 선발(選拔)하기 위하여 한국품종중(韓國品種中)에서 선발(選拔)된 밀양(密陽) 23호(號), 청청(靑靑)벼 등(等) 7품종(品種)과 일본판별품종(日本判別品種)을 각(各) 균주별(菌株別)로 접종(接種) 비교(比較)한 결과(結果) 우리나라 흰잎마름병균에 대(對)한 저항성(抵抗性)과 이병성반응(罹病性反應)이 뚜렷하고 저항성유전자(抵抗性遺傳子)가 다르다고 추정(推定)되는 품종중(品種中)에서 밀양(密陽) 42호(號), 한강(漢江)찰벼, 풍산(豊産)벼, 청청(靑靑)벼, 밀양(密陽) 23호(號)가 한국(韓國) 흰잎마름병균의 병원성(病原性) 변이판별(變異判別)에 적합(適合)한 판별품종(判別品種)으로 선발(選拔)되었다. 우리나라 흰잎마름병 병원균(病原菌)의 병원성(病原性)은 일본판별품종(日本判別品種)의 반응(反應)에 의하여 I, II, III, IV, V 균군(菌群) 등(等)으로 분류(分類)되어 왔으나 한국판별품종(韓國判別品種)의 반응(反應)에 따라 Kl, K2, K3, K4, K5 레이스로 명명(命名)하였다.

• 한국식물병리학회지
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• 제12권2호
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• pp.150-155
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• 1996

1993년과 1994년에 걸쳐 한국의 고추 재배 지역에서 분리한 고추 더뎅이병균은 66개의 균주중 24개가 race 1이었으며, 나머지는 42균주가 race 3이었다. Race 2균주는 발견되지 않았다. 총 66개의 균주중 8%가 200 $\mu\textrm{g}$/ml의 황산구리에 저항성을 보였으며, 100 $\mu\textrm{g}$/ml의 streptomycin sulfate에는 단지 한 균주만이 저항성을 나타냈다. Race 1균주중 21%가 구리에 저항성을 보였으나 race 3균주는 모두 구리에 감수성을 나타냈다. 모든 감수성 균주는 구리 농도 1 $\mu\textrm{g}$/ml에서 죽거나 아주 낮은 수준만이 생존하였으나 저항성 균주는 128 $\mu\textrm{g}$/ml에서도 생존하였다.