• Title/Summary/Keyword: 16S rRNA Pyrosequencing

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Analysis of Soil Bacterial Community in Ihwaryeong and Yuksimnyeong Restoration Project Sites Linking the Ridgeline of Baekdudaegan (이화령 및 육십령 백두대간 생태축 복원사업지 토양 박테리아 군집 분석)

  • Park, Yeong Dae;Kwon, Tae Ho;Eo, Soo Hyung
    • Journal of agriculture & life science
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    • v.50 no.1
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    • pp.117-124
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    • 2016
  • Researches on soil microbial community are increasing to assess ecosystem responses to anthropogenic disturbances and to provide an indicator of ecosystem recovery. Microbial communities are able to respond more rapidly to environmental changes than plants and therefore they may provide an early indication of the ecosystem recovery trajectory. This study was conducted using 16S rRNA gene pyrosequencing of soil samples to compare soil bacterial community composition between artificially covered soils of the Baedudaegan ridgeline and their adjacent forest soils in two restoration project sites, Ihwaryeong and Yuksimnyeong, which were completed in 2012 and 2013, respectively. Richness of the Phylum level was 29.3 in Ihwaryeong and 32.3 in Yuksimnyeong. Significant difference in the richness between artificial restored soils and adjacent forest soils(p<0.01) was observed, however no significant difference was observed for site location and soil depth. Acidobacteria(37.3%) and Proteobacteria(31.1%) were more abundant than any other phylum in collected soil samples. Also, we found the significant difference in the relative abundance of the two abundant phyla between artificially restored soils and their adjacent forest soils (Proteobacteria, 38.1% in restored soils vs 24.2% in adjacent forest soils, p<0.01; Acidobacteria, 55.4% in restored soils vs 19.2% in adjacent forest soils, p<0.001). The results support the previous researches indicating that soil bacterial community composition is affected by nutritional status of soils and that Acidobacteria is also strongly influenced by pH, thus favoring soils with lower pH. This study could be utilized to monitor and evaluate restoration success of forest soil environment quantitatively.

Analysis of Community Structure of Metabolically Active Bacteria in a Rice Field Subjected to Long-Term Fertilization Practices

  • Ahn, Jae-Hyung;Choi, Min-Young;Lee, Hye-Won;Kim, Byung-Yong;Song, Jaekyeong;Kim, Myung-Sook;Weon, Hang-Yeon
    • Korean Journal of Soil Science and Fertilizer
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    • v.46 no.6
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    • pp.585-592
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    • 2013
  • To estimate the effect of long-term fertilization on metabolically active bacterial communities in a rice field, RNA was extracted from endosphere (rice root), rhizosphere, and bulk soil that had been subjected to different fertilization regimes for 59 years, and the 16S rRNAs were analyzed using the pyrosequencing method. The richness and diversity of metabolically active bacteria were higher in bulk soil than in the endosphere and rhizosphere, and showed no significant difference between non-fertilized and fertilized plots. Weighted UniFrac analysis showed that each compartment had characteristic bacterial communities and that the effect of long-term fertilization on the structure of bacterial community was more pronounced in bulk soil than in the endosphere and rhizosphere. The 16S rRNAs affiliated with Alphaproteobacteria and Firmicutes were more abundant in the endosphere than in bulk soil while those affiliated with Chloroflexi and Acidobacteria were more abundant in bulk soil than in the endosphere. Several dominant operational taxonomic units (clustered at a 97% similarity cut-off) showed different frequencies between non-fertilized and fertilized plots, suggesting that the fertilization affected their activities in the rice field.

Analysis of gut bacterial diversity and exploration of cellulose-degrading bacteria in xylophagous insects (목질섭식곤충의 장내 세균 다양성 분석 및 섬유소 분해균 탐색)

  • Choi, Min-Young;Ahn, Jae-Hyung;Song, Jaekyeong;Kim, Seong-Hyun;Bae, Jin-Woo;Weon, Hang-Yeon
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.209-220
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    • 2015
  • In this study, gut bacterial communities in xylophagous insects were analyzed using the pyrosequencing of 16S rRNA genes for their potential biotechnological applications in lignocelluloses degradation. The result showed that operational taxonomic units (OTUs), species richness and diversity index were higher in the hindgut than in the midgut of all insect samples analyzed. The dominant phyla or classes were Firmicutes (54.0%), Bacteroidetes (14.5%), ${\gamma}-Proteobacteria$ (12.3%) in all xylophagous insects except for Rhinotermitidae. The principal coordinates analysis (PCoA) showed that the bacterial community structure mostly clustered according to phylogeny of hosts rather than their habitats. In our study, the two carboxymethyl cellulose (CMC)-degrading isolates which showed the highest enzyme activity were most closely related to Bacillus toyonensis $BCT-7112^T$ and Lactococcus lactis subsp. hordniae $NCDO\;2181^T$, respectively. Cellulolytic enzyme activity analysis showed that ${\beta}-1,4-glucosidase$, ${\beta}-1,4-endoglucanase$ and ${\beta}-1,4-xylanase$ were higher in the hindgut of Cerambycidae. The results demonstrate that xylophagous insect guts harbor diverse gut bacteria, including valuable cellulolytic bacteria, which could be used for various biotechnological applications.

Comparative Analysis of Soil Microbial Communities between Conventional and Organic Farming Systems in Pepper Cultivation (관행과 유기농 고추 재배지의 토양미생물 군집 비교)

  • Kim, Yiseul;Lee, Youngmi;Weon, Hang-Yeon;Sang, Mee Kyung;Song, Jaekyeong
    • Korean Journal of Organic Agriculture
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    • v.28 no.2
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    • pp.235-250
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    • 2020
  • Agricultural practices are known to have a crucial influence not only on soil physico-chemical properties but also on microbial communities. To investigate the effect of farming practices on soil microbial communities, a total of 10 soil samples were collected, including five conventional and five organic farming soils cultivated with peppers in plastic greenhouse. We conducted barcorded-pyrosequencing of V1-V3 regions of 16S rRNA genes to examine soil microbial communities of two different farming practices. Taxonomic classification of the microbial communities at the phylum level indicated that a total of 22 bacterial phyla were present across all samples. Among them, seven abundant phyla (>3%) including Proteobacteria, Actinobacteria, Firmicutes, Acidobacteria, Bacteroidetes, Chloroflexi, and Gemmatimonadetes were found, and Proteobacteria (33.0 ± 5.7%), Actinobacteria (19.9 ± 9.7%), and Firmicutes (13.6 ± 5.0%) comprised more than 66% of the relative abundance of the microbial communities. Organic farming soils showed higher relative abundances of Proteobacteria and Firmicutes, while Actinobacteria and Chloroflexi were more abundant in conventional farming soils. Notably, the genera Bacillus (higher in organic farming soils) and Streptomyces (higher in conventional farming soils) exhibited significant variation in relative abundance between organic and conventional farming soils. Finally, correlation analysis identified significant relationships (p<0.05) between soil chemical properties, in particular, pH and organic matter content and microbial communities. Taken together, this study demonstrated that the changes of soil physico-chemical properties by agricultural farming practices effected significantly (p<0.05) on soil microbial communities.

Changes in Oral Microbiota in Patients Receiving Radical Concurrent Chemoradiotherapy for The Head and Neck Squamous Cell Carcinoma

  • Kim, Jin Ho;Choi, Yoon Hee;An, Soo-Youn;Son, Hee Young;Choi, Chulwon;Kim, Seyeon;Chung, Jin;Na, Hee Sam
    • International Journal of Oral Biology
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    • v.43 no.1
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    • pp.13-21
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    • 2018
  • Radiotherapy (RT) is a mainstay in the treatment of head and neck squamous cell carcinoma (HNSCC). For locally advanced HCSCC, concurrent chemoradiotherapy (CCRT) benefits HCSCC patients in terms of better survival and loco-regional control. In this study, we evaluated changes in oral microbiota in patients, who received CCRT for head and neck cancer. Oral rinsed samples were weekly collected before and during CCRT and at 4 weeks following treatment from HNSCC patients, who had received 70 Gy of radiation delivered to the primary sites for over 7 weeks and concurrent chemotherapy. Oral microbiota changes in three patients were analyzed by next-generation sequencing using 16S rRNA 454 pyrosequencing. On an average, 15,000 partial 16S rRNA gene sequences were obtained from each sample. All sequences fell into 11 different bacterial phyla. During early CCRT, the microbial diversity gradually decreased. In a patient, who did not receive any antibiotics during the CCRT, Firmicutes and Proteobacteria were the most abundant phylum. During the early CCRT, proteobacteria gradually decreased while Firmicutes increased. During the late CCRT, firmicutes gradually decreased while Bacteroides and Fusobacteria increased. In all the patients, yellow complex showed a gradual decrease, while orange and red complex showed a gradual increase during the CCRT. At 4 weeks after CCRT, the recovery of oral microbiota diversity was limited. During CCRT, there was a gradual increase in major periodontopathogens in association with the deterioration of the oral hygiene. Henceforth, it is proposed that understanding oral microbiota shift should provide better information for the development of effective oral care programs for patients receiving CCRT for HNSCC.

Effects of Acarbose Addition on Ruminal Bacterial Microbiota, Lipopolysaccharide Levels and Fermentation Characteristics In vitro

  • Yin, Yu-Yang;Liu, Yu-Jie;Zhu, Wei-Yun;Mao, Sheng-Yong
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.12
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    • pp.1726-1735
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    • 2014
  • This study investigated the effects of acarbose addition on changes in ruminal fermentation characteristics and the composition of the ruminal bacterial community in vitro using batch cultures. Rumen fluid was collected from the rumens of three cannulated Holstein cattle fed forage ad libitum that was supplemented with 6 kg of concentrate. The batch cultures consisted of 8 mL of strained rumen fluid in 40 mL of an anaerobic buffer containing 0.49 g of corn grain, 0.21 g of soybean meal, 0.15 g of alfalfa and 0.15g of Leymus chinensis. Acarbose was added to incubation bottles to achieve final concentrations of 0.1, 0.2, and 0.4 mg/mL. After incubation for 24 h, the addition of acarbose linearly decreased (p<0.05) the total gas production and the concentrations of acetate, propionate, butyrate, total volatile fatty acids, lactate and lipopolysaccharide (LPS). It also linearly increased (p<0.05) the ratio of acetate to propionate, the concentrations of isovalerate, valerate and ammonia-nitrogen and the pH value compared with the control. Pyrosequencing of the 16S rRNA gene showed that the addition of acarbose decreased (p<0.05) the proportion of Firmicutes and Proteobacteria and increased (p<0.05) the percentage of Bacteroidetes, Fibrobacteres, and Synergistetes compared with the control. A principal coordinates analysis plot based on unweighted UniFrac values and molecular variance analysis revealed that the structure of the ruminal bacterial communities in the control was different to that of the ruminal microbiota in the acarbose group. In conclusion, acarbose addition can affect the composition of the ruminal microbial community and may be potentially useful for preventing the occurrence of ruminal acidosis and the accumulation of LPS in the rumen.

The Use of the Pathogen-specific Bacteriophage BCP8-2 to Develop a Rice Straw-derived Bacillus cereus-free Starter Culture (단일 박테리오파지를 이용한 볏짚 유래 Bacillus cereus free 스타터 컬쳐의 개발)

  • Bandara, Nadeeka;Chung, Seo-Jin;Jeong, Do-Youn;Kim, Kwang-Pyo
    • Korean Journal of Food Science and Technology
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    • v.46 no.1
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    • pp.115-120
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    • 2014
  • The purpose of this study was to develop a rice straw-derived Bacillus cereus (B. cereus)-free starter culture for traditional soybean fermented products using a B. cereus-specific bacteriophage, BCP8-2. To determine the optimal medium that supports the growth of rice straw-derived microorganisms and BCP8-2 activity, 5 different culture media were tested. The 5% ground bean (GB) medium was selected for further study. No B. cereus was detected in the BCP8-2-treated rice straw in GB medium, whereas B. cereus at a level of $10^7$ CFU/mL was recovered in the no-phage control. The total bacterial count reached approximately $10^9$ CFU/mL regardless of phage addition. When the 16S rRNA sequence-based microbial community was monitored using denaturing gradient gel electrophoresis (DGGE) and pyrosequencing, a similar microbial community was observed in the phage-treated and control samples. In conclusion, we demonstrate that phage can be used to prepare a rice straw-derived B. cereus-free starter culture with minimal effect on natural microflora.

PD-1 deficiency protects experimental colitis via alteration of gut microbiota

  • Park, Seong Jeong;Kim, Ji-Hae;Song, Mi-Young;Sung, Young Chul;Lee, Seung-Woo;Park, Yunji
    • BMB Reports
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    • v.50 no.11
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    • pp.578-583
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    • 2017
  • Programmed cell death-1 (PD-1) is a coinhibitory molecule and plays a pivotal role in immune regulation. Here, we demonstrate a role for PD-1 in pathogenesis of inflammatory bowel disease (IBD). Wild-type (WT) mice had severe wasting disease during experimentally induced colitis, while mice deficient for PD-1 ($PD-1^{-/-}$) did not develop colon inflammation. Interestingly, $PD-1^{-/-}$ mice cohoused with WT mice became susceptible to colitis, suggesting that resistance of $PD-1^{-/-}$ mice to colitis is dependent on their gut microbiota. 16S rRNA gene-pyrosequencing analysis showed that $PD-1^{-/-}$ mice had altered composition of gut microbiota with significant reduction in Rikenellaceae family. These altered colon bacteria of $PD-1^{-/-}$ mice induced less amount of inflammatory mediators from colon epithelial cells, including interleukin (IL)-6, and inflammatory chemokines. Taken together, our study indicates that PD-1 expression is involved in the resistance to experimental colitis through altered bacterial communities of colon.

Assessment of Microbial Community in Paddy Soils Cultivated with Bt and Nakdong Rice (Bt 벼의 토양미생물상 영향 비교평가)

  • Sohn, Soo-In;Ahn, Byung-Ohg;Chi, Hee-Youn;Cho, Byung-Kwan;Cho, Min-Seok;Shin, Kong Sik
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.5
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    • pp.829-835
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    • 2012
  • The cultivation of genetically modified (GM) crops has increased due to their economic and agronomic advantages. Before commercialization of GM crops, however, we must assess the potential risks of GM crops on human health and environment. The aim of this study was to investigate the possible impact of Bt rice on the soil microbial community. Microbial communities were isolated from the rhizosphere soil cultivated with Bt rice and Nakdong, parental cultivar and were subjected to be analyzed using both culture-dependent and molecular methods. The total counts of bacteria, fungi, and actinomycetes in the rhizosphere of transgenic and conventional rice were not significantly different. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes revealed that the bacterial community structures during cultural periods were very similar each other. Analysis of dominant isolates in the rhizosphere cultivated with Bt and Nakdong rice showed that the dominant isolates from the soil of Bt rice and Nakdong belonged to the Proteobacteria, Cloroflexi, Actinobacteria, Firmicutes, and Acidobacteria. These results indicate that the Bt rice has no significant impact on the soil microbial communities during cultivation period. Further study remains to be investigated whether the residue of Bt rice effect on the soil environment.

Soil Bacterial Community in Red Pine Forest of Mt. Janggunbong, Bonghwa-Gun, Gyeongbuk, Korea, Using Next Generation Sequencing (차세대염기서열방법을 이용한 경북 봉화군 장군봉 소나무림의 토양 박테리아 군집 구성)

  • Lee, Byeong-Ju;Eo, Soo Hyung
    • Journal of Korean Society of Forest Science
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    • v.106 no.2
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    • pp.121-129
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    • 2017
  • The soil microbiome plays important roles in material cycling and plant growth in forest ecosystem. Although a lot of researches on forest soil fungi in Korea have been performed, the studies on forest soil bacterial communities have been limited. In this study, we conducted next generation sequencing (NGS) targeting 16S rRNA gene to investigate the soil bacterial communities from natural red pine (Pinus densiflora) forest in Mt. Janggunbong, Bonghwa-gun, Gyeongbuk, Korea. Our results showed that the entire bacterial communities in the study sites include the phyla Proteobacteria, Acidobacteria, Actinobacteria, Planctomycetes, which have been typically observed in forest soils. The composition ratio of Proteobacteria was the highest in the soil bacteria community. The results reflect that Proteobacteria is copiotroph, which generally favors relatively nutrient-rich conditions with abundant organic matter. Some rhizobia species such as Burkholderia, Bradyrhizobium, Rhizobium, which are known to contribute to soil nitrogen-fixation, exist in the study sites. As a result of correlation analysis between soil physicochemical characteristics and bacteria communities, the soil pH was significantly correlated with the soil bacteria compositions.