• Title/Summary/Keyword: Bacterial DNA

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Effect of Korean Fermented Food Extracts and Bacteria Isolated from the Extracts for the Control of Rice Seed-borne Fungal Diseases (국산 발효식품 추출물과 발효식품유래 미생물을 활용한 벼 종자전염성 진균병 방제)

  • Kim, Yong-Ki;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Lee, Min-Ho;Park, Jong-Ho;Han, Eun-Jung;Choi, Eun-Jung;Bae, Soo-Il;Jee, Hyeong-Jin
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.383-395
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    • 2014
  • When we investigated seed infestation by fungal pathogens from 51 varieties in 9 crops, the contamination rate of rice and sesame seeds was high. Therefore, to control seed-borne diseases, we obtained extracts from commercial products of Kimchi, Gochujang, Doenjang, Ganjang, Makgeolli and Tohajut and their suppressive effects against seed-borne diseases were studied. In addition, bacterial strains were screened to control rice seed-borne diseases in vitro and in vivo. Among forty food extracts, eleven food-extracts suppressed incidence of seedling rots in vitro and five food extracts increased 8-33% of healthy seedling in the greenhouse. Among 218 isolates from 40 fermented foods, 43 isolates showed high antifungal activity against seven fungal pathogens. When we tested 43 isolates for the reduction of rice seed borne disease, 32 isolates were able to reduce the rice seed borne disease. Among 32 isolates, 17 isolates reduced significantly seedling rot and increased healthy seedlings, the other isolates except for Kc4-2 and Mkl 2-2 increased shoot emergence and the percentage of healthy plants. Thirty isolates with high antifungal activity and suppressive effect against rice seedling rots were identified by 16S rRNA sequencing. Twenty one of thirty isolates were identified as Bacillus spp. Three isolates from Makgeolli were identified as Saccharomyces cerevisiae. B. amyloliquifaciens were isolated from six Korean traditional fermented foods except for Ganjang. B. amyloliquifaciens were majority in the effective bacterial population of Gochujang and Jutgal. Relatively diverse Bacillus species including B. subtilis, B. pumilus, and B. amyloliquificiens were isolated from Kimchi. The selected effective microorganisms from Korean fermented foods founded to be effective for controlling seed-borne diseases of rice in vitro and in the greenhouse. We think that Korean fermented foods and their useful microorganisms can be used as biocontrol agents for suppressing rice seed-borne diseases based on above described results.

Changes of Nitrifying Bacteria Depending on the Presence and Absence of Organic Pollutant in Nak-Dong River (낙동강에서의 유기성 오염 유무에 따른 질화세균의 변화)

  • Jin, Seon-Yeong;Lee, Young-Ok
    • Korean Journal of Microbiology
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    • v.49 no.2
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    • pp.137-145
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    • 2013
  • This study was performed at 2 sites of Nak-Dong River to investigate the changes of nitrifiers depending on the presence and absence of organic pollutants (due to the effluents of domestic wastewater treatment plant, WWTP). Conventional chemical parameters such as T-N, $NH_4$-N, $NO_2$-N, $NO_3$-N were measured and the quantitative nitrifiers at the 2 sites were analyzed comparatively by fluorescent in situ hybridization (FISH) with NSO190 and NIT3, after checking the presence of gene amoA of ammonia oxidizing bacteria (AOB) and 16S rDNA signature sequence for Nitrobacter sp. that belongs to nitrite oxidizing bacteria (NOB). Also ${\alpha}{\cdot}{\beta}{\cdot}{\gamma}$-Proteobacteria were detected using FISH to get a glimpse of the general bacterial community structure of the sites. Based on the distribution structure of the ${\alpha}{\cdot}{\beta}{\cdot}{\gamma}$-Proteobacteria and the measurement of nitrogen in different phases, it could be said that the site 2 was more polluted with organics than site 1. Corresponding to the above conclusion, the average numbers of AOB and NOB detected by NSO160 and NIT3, respectively, at site 2 [AOB, $9.3{\times}10^5$; NOB, $1.6{\times}10^6$ (cells/ml)] was more than those at site 1 [AOB, $7.8{\times}10^5$; NOB, $0.8{\times}10^6$ (cells/ml)] and also their ratios to total counts were higher at site 2 (AOB, 27%; NOB, 34%) than those at site 1 (AOB, 18%; NOB, 23%). Thus, it could be concluded that the nitrification at site 2 was more active due to continuous loading of organics from the effluents of domestic WWTP, compared to site 1 located closed to raw drinking water supply and subsequently less polluted with organics.

Effects of Detoxified Sulfur as a Feed Supplement on in Vitro Rumen Fermentation and Methane Mitigation (제독 유황의 반추위 발효성상 및 메탄 저감 효과 연구)

  • Kim, Seon-Ho;Islam, Mahfuzul;Biswas, Ashraf Ali;Cho, Kwang-Keun;Lee, Sang-Suk
    • Journal of Life Science
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    • v.30 no.9
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    • pp.743-748
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    • 2020
  • Sulfate is a reductant that competes for electrons and may lower CH4 production in the rumen. This study was designed to evaluate the beneficial effect of detoxified sulfur powder supplementation on in vitro rumen fermentation and methane mitigation. A ruminally cannulated Holstein Friesian cow was used as a rumen fluid source, and commercial pelleted concentrate was used as a substrate at 1 g dry matter. Treatments included the addition of detoxified sulfur powder at the rate of 0% (Control), 0.2% (T1), 0.4% (T2), 0.6% (T3), 0.8% (T4), and 1.0% (T5) as dry matter (DM) basis. The pH, total gas (TG), methane (CH4) production, DM digestibility, organic matter (OM) digestibility, and volatile fatty acids (VFA) production were analyzed after 12 hr of incubation. The results showed that CH4 production was significantly lowest in T1 (13.78 ml) but highest in the control (20.16 ml). Insignificantly higher total VFA was observed in control and T1 (64.99 and 64.28 mM, respectively) compared to other treatments after 12 hr of incubation. After 12 hr of incubation, the significantly lowest acetate:propionate was observed in T1 (1.90) while the highest was observed in T4 (2.44). However, no significant differences were recorded for pH, TG, DM digestibility, OM digestibility, acetate, propionate, and butyrate between the control and T1. Total number of bacterial DNA copies was significantly lower in the treatment group than the control. Therefore, it can be concluded from this study that detoxified sulfur at 0.2% inclusion level is optimal for production performance and ruminal CH4 mitigation.

Analysis of Microbial Community Structure in Soil and Crop Root System II. Analysis of soil microbial community structure in different soil Environmental conditions by MIDI and DNA analyses (토양과 작물근계의 미생물군집 구조 해석 II. MIDI 및 DNA 분석에 의한 토양환경별 미생물 군집 해석)

  • Ryu, Jin-Chang;Kwon, Soon-Wo;Kim, Jong-Shik;Suh, Jang-Sun;Jung, Beung-Gan;Choi, Sun-Shik
    • Korean Journal of Soil Science and Fertilizer
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    • v.35 no.2
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    • pp.118-126
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    • 2002
  • To evaluate the correlations of microbial populations with soil healthiness and crop production and establish the criteria for microbial population of soil types. We analyzed the microbial community structure of 13 soils which were different in physical and chemical properties and cultivation methods. According to the analysis of microbial population suing the dilution plate method, the large differences of the microbial population structures among soil types were shown: aerobic bacteria $2-27{\times}10^6$, fluorescent Pseudomonas $1-1,364{\times}10^5$, Gram negative bacteria $1-126{\times}10^4$, and mesophilic Bacillus $1-110{\times}10^5$. The density of Gram negative bacteria was highest on red pepper cultivating soils (sample no. 4 and 6) of Umsung and Gesan, Chungbuk, and the density of the fluorescent Pseudomonas was highest on greenhouse soil (sample no. 7) of Jinju, Kyungnam. The crop productivity of three soils was high as compared with those of other soils. It was supposed that the density of fluorescent Pseudomonas and mesophilic Bacillus were correlated with the incresed crop production. By MIDI analysis, 579 strains isolated from 13 soils composed of a variety of microbes including 102 isolates of Agrobacterium, 112 isolates of Bacillus, 32 isolates of Pseudomonas, 44 isolates of Kocuria, and 34 isolates of Pseudomonas. Among the 624 isolates of Gram negative bacteria, Pseudomonas including P. putida and p. fluorescens occupied the highest density (51%), and Stenotrophomonas maltophilia and Burkholderia cepacia also appeared at high density. From RAPD analysis, the fluorescent Pseudomonas strains isolated from 13 soil types showed a high level of strain diversities and were grouped into 2 - 14 patterns according to soil types. Many of unknown bacteria were recovered from the paddy soil, and needed to be further characterized on the molecular basis.

In vivo Antifungal Activity of Pyrrolnitrin Isolated from Burkholderia capacia EB215 with Antagonistic Activity Towards Colletotrichum Species (탄저병균에 대하여 길항작용을 보이는 Burkholderia cepacia EB215로부터 분리한 Pyrrolnitrin의 항균활성)

  • Park, Ji-Hyun;Choi, Gyung-Ja;Lee, Seon-Woo;Jang, Kyoung-Soo;Choi, Yong-Ho;Chung, Young-Ryun;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Korean Journal of Mycology
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    • v.32 no.1
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    • pp.31-38
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    • 2004
  • An endophytic bacterial strain EB215 that was isolated from cucumber (Cucumis sativus) roots displayed a potent in vivo antifungal activity against Colletotrichum species. The strain was identified as Burkholderia cepacia based on its physiological and biochemical characteristics, and 16S rDNA gene sequence. Optimal medium and incubation period for the production of antifungal substances by B. cepacia EB215 were nutrient broth (NB) and 3 days, respectively. An antifungal substance was isolated from the NB cultures of B. cepacia EB215 strain by centrifugation, n-hexane partitioning, silica gel column chromatography, preparative TLC, and in vitro bioassay. Its chemical structure was determined to be pyrrolnitrin by mass and NMR spectral analyses. Pyrrolnitrin showed potent disease control efficacy of more than 90% against pepper anthracnose (Colletotrichum coccodes), cucumber anthracnose (Colletotrichum orbiculare), rice blast (Magnaporthe grisea) and rice sheath blight (Corticium sasaki) even at a low concentration of $11.1\;{\mu}g/ml$. In addition, it effectively controlled the development of tomato gray mold (Botrytis cinerea) and wheat leaf rust (Puccinia recondita) at concentrations over $33.3\;{\mu}g/ml$. However, it had no antifungal activity against Phytophthora infestans on tomato plants. Further studies on the development of microbial fungicide using B. cepacia EB215 are in progress.

Isolation and Characterization of Exopolysaccharide Producing Lactic Acid Bacteria from Korean Soy Sauce and Soybean Paste (전통 장류로부터 Exopolysaccharide 생성 유산균의 분리 및 특성)

  • Yun, Hye Ju;Lee, You Jung;Yeo, Soo-Hwan;Park, Hye Young;Park, Heui-Dong;Baek, Seong Yeol
    • Microbiology and Biotechnology Letters
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    • v.41 no.2
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    • pp.190-197
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    • 2013
  • Three slime-forming lactic acid bacteria were isolated from traditional Korean fermented soy sauce and soybean paste and shown to produce exopolysaccharides (EPS) in sucrose media. By isolating the strains, examining their morphological characteristics and determining their 16S rDNA sequences, N58-5 and K6-7 were identified as Leuconostoc mesenteroides and N45- 10 as Leuconostoc citreum. The acid and bile tolerances of these three strains were investigated. Amongst the three lactic acid bacteria, Leuc. citreum N45-10 exhibited the highest viability ($10^5-10^6$ CFU/ml) in 0.05 M sodium phosphate buffer (pH 0.3) for 2 h, in artificial gastric juice for 2 h and in 0.3%, 0.5% oxgall for 24h. Leuc. mesenteroides K6-7, N58-5 and Leuc. citreum N45- 10 were grown in sucrose liquid medium and 8.16 g/L, 3.65 g/L, 16.17 g/L of EPS was collected, respectively. The hydrolyzed EPS was analyzed by HPLC in order to determine the sugar composition of EPS. Leuc. mesenteroides K6-7 and N58-5 showed two peaks indicating glucose and fructose, thus they were determined to be hetero-type polysaccharides. Leuc. citreum N45-10 showed only the glucose polymer, indicating it to be a homo-type polysaccharide. In addition, all three lactic acid bacterial hemolysis did not demonstrate a clear zone in blood agar in the area surrounding a lactic acid bacteria colony.

A Domestic Outbreak of Bacterial Dysentery Caused by Extended-Spectrum β-Lactamase(ESBL)-producing Shigella sonnei (집단 발생한 Extended-spectrum β-lactamase(ESBL) 생산 Shigella sonnei 감염에 의한 세균성 이질에 관한 연구)

  • Lim, Hyun Taek;Lee, So Hee;Lee, Jung Hwa;Kim, Jeong Eun;Kim, Kyo Sun;Jeong, Eun Ju;Lee, Seung Hyun;Kang, Chang Gyu;Hong, Seong Jin
    • Clinical and Experimental Pediatrics
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    • v.48 no.10
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    • pp.1107-1115
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    • 2005
  • Purpose : An outbreak of ESBL-producing Shigella sonnei enteritis was unprecedented not only in Korea but throughout the world in the past. We intended to devise a management guideline for ESBL-producing shigellosis based on analysis of clinical manifestations and response to therapy. Methods : We analyzed 103 patients who were admitted to the hospital with acute GI symptoms and were shown positive result for S. sonnei on stool culture. We performed sensitivity test to the antibiotics and DNA sequencing of ESBL gene in the isolated S. sonnei colonies. In addition, we retrospectively analyzed their clinical characteristics, laboratory results, and clinical and microbiological responses to the antibiotics. Results : Among the clinical manifestations, fever was the most frequent(96.1%), followed by diarrhea(93.2%), abdominal pain(76.7%), headache(71.8%), vomiting(65.0%), and nausea(41.7%). The fever was sustained for average of 2.0 days and diarrhea for 3.9 days. Watery diarrhea was the most common(69%) followed by mucoid(26%), and bloody stool(5%). On peripheral blood smear, leukocytosis was noted in 53.4% of patients, and 78.6% of patients tested positive for serum CRP response. On stool direct smear, 11.7% of patients showed more than 50 WBCs/HPF, and 9.7% of patients between 5 to 20 WBCs/HPF. Stool occult blood was positive in 71% of patients. Production of CTX-M-14 type ESBL was reported for all S. sonnei strains isolated from this outbreak. Microbiological eradication rates to various antibiotics were as follows : 100%(9/9) to ciprofloxacin, 100% 5/5) to azithromycin, 6.9%(5/72) to cefdinir, 0%(0/8) to ceftriaxone, 12.5%(1/8) to ceftizoxime, 0%(0/ 8) to TMP/SMX, 42.9%(3/7) to ampicillin/sulbactam, 20%(1/5) to amoxicillin/clavulanic acid, and 68.8 %(11/16) to imipenem/cilastatin. Conclusion : It is presumed that azithromycin can be an attractive option for the treatment of ESBL-producing S. sonnei enteritis in pediatric population, given its cost-effectiveness and safety. Although ciprofloxacin is another cost-effective agent, its use in pediatric population may be a bit too premature.

Identification of Novel Psychrotolerant Bacterial Strain and Production of $\beta-Galactosidase$ (새로운 저온 내성세균의 동정과 $\beta-Galactosidase$ 생산)

  • Park, Jeong-Woon;Yoo, Jae-Soo;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.40-46
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    • 2006
  • Galactose joined to glucose by a $\beta(1\rightarrow4)$ glycosidic bond makes lactose and this disaccharide is rich in milk. It is known that lacotse is hydrolyzed to each monomeric sugar by either lactase in human or $\beta-galactosidase$ in bacteria. Ingestion of milk by lactase-deficient persons causes a temporary diarrhea and subsequent chronic diarrhea results in colitis with chronic inflammation. We isolated a $\beta-galactosidase$ producing psycrotolerant strain AS-20 from near cattle shed and investigated the growth at various temperature conditions. Whereas Escherichia coli strains did not grow at $10^{\circ}C$, the AS-20 strain could grow well at this low temperature and showed optimal growth at $30^{\circ}C$. The isolated strain was identified as 97% Hafnia alvei by biochemical properties. This strain could ferment glucose, lacotse, maltose, mannitol, xylose, ONPG, rhamanose and L-arabinose, and decarboxylate lysin and ornithine. To confirm the identity of isolated strain we amplified 16S rDNA by PCR and searched similarity of the 1426 bp DNA sequcence with Genbank database. The strain AS-20 showed 99% similarity with Hafnia alvei. The activity of $\beta-galactosidase$ was 1.5 times higher when the cell was grown at 10 or $20^{\circ}C$ than at $30^{\circ}C$. The highest enzyme activity of AS-20 was also much higher than that of E. coli, which was grown at $30^{\circ}C$.

Development of dry milling suitable rice cultivar to invigorate rice processing products

  • Jeung, Ji-Ung
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.10-10
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    • 2017
  • Rice consumption has been continuously decreasing as the eating habits of Koreans have become westernized and diversified. The per capita annual rice consumption in Korea has dropped sharply from 136.4 kg in 1970 to 61.9 kg in 2016. The Korean government, therefore, has been trying to promote rice consumption by invigorating the processed food industry using rice flour. To facilitate the market for processed rice foods, it is essential to develop proper milling technology in terms of flour particle size and damaged starch content to produce high quality rice flour at competitive cost. Dry milling and wet milling are the two major processes used to produce rice flour. Although the dry milling process is relatively simple with a lower production cost, damaged starch content increases because of the high grain hardness of rice. In wet milling, the quality of rice flour is improved by reducing flour particle size as well as damaged starch content through soaking procedures. However, the production costs are high because of the additional expenses associated with the disposal of waste water, sterilization and drying of the wet flour. Recently developed technologies such as jet milling and cryogenic milling also require expensive investment and production. Therefore, developing new rice cultivars with dry milling adaptability as well as good processing properties is an important goal of rice breeding in Korea. 'Suweon 542' is a floury endosperm mutant line derived from sodium azide treatment on a high-yield, early maturing, and non-glutinous japonica rice cultivar, 'Namil'. Compared with the wild type, after dry milling process, the grain hardness of 'Suweon 542' was significantly lower because of its round and loosely packed starch granules. Also, the flour of 'Suweon 542' had significantly smaller particles and less damaged starch than 'Namil' and other rice cultivars and its particle size distribution was similar to a commercial wheat cultivar. Recently, through collaborations with nine universities and food companies, a total of 21 kinds of processed prototypes, using the dry milling flour of 'Suweon 542', were evaluated. In the production of major rice processing products, there was no significant quality difference between the flours prepared by wet milling and dry milling. Although the amount of water added to the dough was slightly increased, it was confirmed that the recipe applying the wet flour could be used without significant change. To efficiently transfer the floury endosperm characteristics of 'Suweon 542' to other commercial rice cultivars, it is essential to develop DNA marker tightly linked to the target gene. Association analysis using 70 genome-wide SSR markers and 94 F2 plants derived from 'Suweon 542'/'Milyang 23' showed that markers on chromosome 5 explained a large portion of the variation in floury grains percentage (FGP). Further analysis with an increased number of SSR markers revealed that the floury endosperm of 'Suweon 542' was directed by a major recessive locus, flo7(t), located in the 19.33-19.86 Mbp region of chromosome 5, with RM18639 explaining 92.2% of FGP variation in the F2 population. Through further physical mapping, a co-segregate and co-dominant DNA marker with the locus, flo7(t) was successfully developed, by which, thereby, breeding efficiency of rice cultivars having proper dry milling adaptability with high yield potential or useful functional materials would be improved. 'Suweon 542' maintained the early maturity of the wild type, Namil, which can be used in rice-wheat double cropping systems in Korea not only for improved arable land but also for sharing flour production facilities. In addition to the high susceptibility against major rice diseases, nevertheless, another possible drawback of 'Suweon 542' is the high rate of viviparous under prolonged rainfall during the harvesting season. To overcome susceptibility and vivipary of 'Suweon 542', the progeny lines, derived from the crosses 'Suweon 542' and 'Jopyeong', an early maturing rice cultivar with multiple resistance against rice blast, bacterial blight, and rice strip virus, and 'Heugjinju', a anthocyanin pigment containing black rice cultivar, were intensively evaluated. As the outputs, three dry milling suitable rice elite lines, 'Jeonju614', 'Jeonju615', and 'Jeonju616' were developed.

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Comparison of Group A, B and C Rotaviral Gastroenteritis among Children in Korea: Prevalence and Clinical Features (한국 소아에서의 로타 장염군의 비교: 유병율과 임상증상)

  • Bae, Kil-Seong;Bae, Woo Ri;Kim, Ji Hoon;Bin, Joong Hyun;Kim, Hyun Hee;Lee, Hee Jin;Lee, Wonbae
    • Pediatric Infection and Vaccine
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    • v.21 no.2
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    • pp.96-103
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    • 2014
  • Purpose: The aim of this study is that the prevalence of rotavirus infection was evaluated by each group and clinical features of group A, B and C rotaviruses infections were described respectively to compare one with another. Methods: Between January 2010 and December 2010, we enrolled a group of children below 10 years of age admitted for management of acute diarrhea at the Catholic University of Korea Bucheon St. Mary's Hospital. A total of 310 stool samples documented to be free of common bacterial pathogens were collected from children with diarrhea. The presence of group A, B or C rotavirus is indicated by amplification of DNA segments of the expected lengths after the first and second PCRs Results: In a total of 310 stool specimens, 40 (12.9%) specimens were positive for rotaviruses. These included 23 (7.4%) positive for group A, 5 (1.6%) for group B and 12 (3.9%) for group C rotaviruses. Group B rotavirus infected patients had significantly less diarrheas per day (group A: P =0.01, group C: P =0.01) and shorter duration of vomiting days (group A: P =0.03, group C: P =0.03) than those with group A and C rotaviruses infection respectively. All the group B rotaviruses had been isolated in March and October. Group C rotavirus infections were prevalent during late summer and early winter and peaked in October. Conclusion: These findings indicate that group B and C rotaviruses are notable causes or the contributing causes of diarrhea among infants and children in Korea.