• 제목/요약/키워드: miSeq

검색결과 106건 처리시간 0.023초

16S rRNA 유전자 서열 분석을 이용한 DNA 및 cDNA 기반 장내 미생물 군집 분석의 비교 (Comparison between DNA- and cDNA-based gut microbial community analyses using 16S rRNA gene sequences)

  • 조혜준;홍지완;운노타쯔야
    • 미생물학회지
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    • 제55권3호
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    • pp.220-225
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    • 2019
  • 최근 10년간 미생물생태분석 기반의 연구는 차세대염기서열분석법이 개발된 이래로 지속적으로 증가하고 있다. 장내미생물생태와 건강의 연관성은 미생물 생태학 분야에 있어서 중요한 결과로 여겨지고 있다. 미생물 군집 분석은 주로 16S rRNA 유전자 가변 영역의 염기서열을 통해 분석되지만 이는 미생물의 활성 정보를 제공하지 않는다. 본 연구에서는 cDNA 기반의 미생물 생태분석을 수행하고 DNA 및 cDNA기반의 미생물생태분석 결과를 비교하였다. 두 가지의 서로 다른 접근법이 Butyrate producer와 probiotics와 같이 장내 대사과정에서 중요한 미생물의 abundance 뿐만 아니라 비만 지표로 알려진 Firmicutes 와 Bacteroidetes의 비율에 있어서 차이가 있음을 나타내었다. 따라서, cDNA 기반 미생물 군집은 이전에 수행된 DNA 기반 미생물 군집 분석과 비교하여 장내미생물생태의 역할과 관련된 또 다른 분석 방향성을 제공한다.

Responses of Soil Bacterial and Fungal Communities to Organic and Conventional Farming Systems in East China

  • Zhang, Hanlin;Zheng, Xianqing;Bai, Naling;Li, Shuangxi;Zhang, Juanqin;Lv, Weiguang
    • Journal of Microbiology and Biotechnology
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    • 제29권3호
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    • pp.441-453
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    • 2019
  • Organic farming is considered an effective form of sustainable agricultural management. However, understanding of soil microbial diversity and composition under long-term organic and conventional farming is still limited and controversial. In this study, the Illumina MiSeq platform was applied to investigate the responses of soil bacterial and fungal diversity and compositions to organic farming (OF) and improved conventional farming (CF, applied straw retention) in the rice-wheat rotation system. The results highlighted that the alpha diversity of microbial communities did not differ significantly, except for higher bacterial diversity under OF. However, there were significant differences in the compositions of the soil bacterial and fungal communities between organic and conventional farming. Under our experimental conditions, through the ecological functional analysis of significant different or unique bacterial and fungal taxonomic members at the phyla and genus level, OF enhanced nitrogen, sulfur, phosphorus and carbon dynamic cycling in soil with the presence of Nodosilinea, Nitrospira, LCP-6, HB118, Lyngbya, GOUTA19, Mesorhizobium, Sandaracinobacter, Syntrophobacter and Sphingosinicella, and has the potential to strengthen soil metabolic ability with Novosphingobium. On the other hand, CF increased the intensity of nitrogen cycling with Ardenscatena, KD1-23, Iamia, Nitrosovibrio and Devosia, but enriched several pathogen fungal members, including Coniochaeta, Corallomycetella, Cyclaneusma, Cystostereum, Fistulina, Curvularia and Dissoconium.

Comparative Transcriptomic Analysis of MAPK-Mediated Regulation of Sectorization in Cryphonectria parasitica

  • Chun, Jeesun;So, Kum-Kang;Ko, Yo-Han;Kim, Jung-Mi;Kim, Dae-Hyuk
    • Molecules and Cells
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    • 제42권4호
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    • pp.363-375
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    • 2019
  • Fungal sectorization is a complex trait that is still not fully understood. The unique phenotypic changes in sporadic sectorization in mutants of CpBck1, a mitogen-activated protein kinase kinase kinase (MAPKKK) gene, and CpSlt2, a mitogen-activated protein kinase (MAPK) gene, in the cell wall integrity pathway of the chestnut blight fungus Cryphonectria parasitica have been previously studied. Although several environmental and physiological factors cause this sectoring phenotype, genetic variants can also impact this complex morphogenesis. Therefore, RNA sequencing analysis was employed to identify candidate genes associated with sectorization traits and understand the genetic mechanism of this phenotype. Transcriptomic analysis of CpBck1 and CpSlt2 mutants and their sectored progeny strains revealed a number of differentially expressed genes (DEGs) related to various cellular processes. Approximately 70% of DEGs were common between the wild-type and each of CpBck1 and CpSlt2 mutants, indicating that CpBck1 and CpSlt2 are components of the same MAPK pathway, but each component governs specific sets of genes. Functional description of the DEGs between the parental mutants and their sectored progenies revealed several key pathways, including the biosynthesis of secondary metabolites, translation, amino acid metabolism, and carbohydrate metabolism; among these, pathways for secondary metabolism and translation appeared to be the most common pathway. The results of this comparative study provide a better understanding of the genetic regulation of sector formation and suggest that complex several regulatory pathways result in interplays between secondary metabolites and morphogenesis.

Effect of Bacillus mesonae H20-5 Treatment on Rhizospheric Bacterial Community of Tomato Plants under Salinity Stress

  • Lee, Shin Ae;Kim, Hyeon Su;Sang, Mee Kyung;Song, Jaekyeong;Weon, Hang-Yeon
    • The Plant Pathology Journal
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    • 제37권6호
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    • pp.662-672
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    • 2021
  • Plant growth-promoting bacteria improve plant growth under abiotic stress conditions. However, their effects on microbial succession in the rhizosphere are poorly understood. In this study, the inoculants of Bacillus mesonae strain H20-5 were administered to tomato plants grown in soils with different salinity levels (EC of 2, 4, and 6 dS/m). The bacterial communities in the bulk and rhizosphere soils were examined 14 days after H20-5 treatment using Illumina MiSeq sequencing of the bacterial 16S rRNA gene. Although the abundance of H20-5 rapidly decreased in the bulk and rhizosphere soils, a shift in the bacterial community was observed following H20-5 treatment. The variation in bacterial communities due to H20-5 treatment was higher in the rhizosphere than in the bulk soils. Additionally, the bacterial species richness and diversity were greater in the H20-5 treated rhizosphere than in the control. The composition and structure of the bacterial communities varied with soil salinity levels, and those in the H20-5 treated rhizosphere soil were clustered. The members of Actinobacteria genera, including Kineosporia, Virgisporangium, Actinoplanes, Gaiella, Blastococcus, and Solirubrobacter, were enriched in the H20-5 treated rhizosphere soils. The microbial co-occurrence network of the bacterial community in the H20-5 treated rhizosphere soils had more modules and keystone taxa compared to the control. These findings revealed that the strain H20-5 induced systemic tolerance in tomato plants and influenced the diversity, composition, structure, and network of bacterial communities. The bacterial community in the H20-5 treated rhizosphere soils also appeared to be relatively stable to soil salinity changes.

Composition and functional diversity of bacterial communities during swine carcass decomposition

  • Michelle Miguel;Seon-Ho Kim;Sang-Suk Lee;Yong-Il Cho
    • Animal Bioscience
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    • 제36권9호
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    • pp.1453-1464
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    • 2023
  • Objective: This study investigated the changes in bacterial communities within decomposing swine microcosms, comparing soil with or without intact microbial communities, and under aerobic and anaerobic conditions. Methods: The experimental microcosms consisted of four conditions: UA, unsterilized soil-aerobic condition; SA, sterilized soil-aerobic condition; UAn, unsterilized soil-anaerobic condition; and San, sterilized soil-anaerobic condition. The microcosms were prepared by mixing 112.5 g of soil and 37.5 g of ground carcass, which were then placed in sterile containers. The carcass-soil mixture was sampled at day 0, 5, 10, 30, and 60 of decomposition, and the bacterial communities that formed during carcass decomposition were assessed using Illumina MiSeq sequencing of the 16S rRNA gene. Results: A total of 1,687 amplicon sequence variants representing 22 phyla and 805 genera were identified in the microcosms. The Chao1 and Shannon diversity indices varied in between microcosms at each period (p<0.05). Metagenomic analysis showed variation in the taxa composition across the burial microcosms during decomposition, with Firmicutes being the dominant phylum, followed by Proteobacteria. At the genus level, Bacillus and Clostridium were the main genera within Firmicutes. Functional prediction revealed that the most abundant Kyoto encyclopedia of genes and genomes metabolic functions were carbohydrate and amino acid metabolisms. Conclusion: This study demonstrated a higher bacteria diversity in UA and UAn microcosms than in SA and SAn microcosms. In addition, the taxonomic composition of the microbial community also exhibited changes, highlighting the impact of soil sterilization and oxygen on carcass decomposition. Furthermore, this study provided insights into the microbial communities associated with decomposing swine carcasses in microcosm.

Candidate Genes Related to Sugar Content in Sweetpotato using GWAS

  • Tae Hwa Kim;Mi Nam Chung;Hyeong Un Lee;Won Park;Sang Sik Nam
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2022년도 추계학술대회
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    • pp.192-192
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    • 2022
  • Sweetpotato is rich in starch, which is converted to sugar during storage due to enzymatic hydrolysis. The sugar content of sweetpotato is a component related to taste and storability. In this study, the sugar content (fructose, glucose, maltose, sucrose and total sugar content) of 94 genotypes was evaluated and the GWAS (Genome-Wide Association Study) was conducted to search for candidate genes for sugar content. The fructose and glucose content were 0.2 ~ 8.8 and 0.2 ~ 9.4 g/100g, respectively. The maltose, sucrose and total sugar content were 0.2 ~ 9.1,3.2 - 30.0 and 7.9 ~ 40.2 g/100g, respectively. The fructose and glucose showed a positive correlation (0.98). The 94 genotypes were genotyped with genotyping-by-sequencing (GBS) and aligned against the reference genome sequences of sweetpotato. The GBS libraries from 94 genotypes were sequenced on an Illumina HiSeqXten system, and 1,339,892 SNPs (Single Nucleotide Polymorphism) were generated. Filtering for < 60% missing rate and > 0.05 minor allele frequency resulted in a total of 44,255 SNPs used in GWAS. The GAPIT (Genome Association and Prediction Integrated Tool) was used to conduct based on the mean of sugar content with a Bonferroni-corrected chromosome-wide significance threshold with a -logio(P) of 5.95. The significant SNPs were obtained with fructose (seven), glucose (six), maltose (four) and sucrose (nine). There were several genes related to sugar content around the significant SNPs such as sugar transport protein 8-like, probable galactose-1 -phosphate uridyltransferase-like and beta-amylase. These results will contribute to understanding of sugar content and conversion in sweetpotato.

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Combined Application Effects of Arbuscular Mycorrhizal Fungi and Biochar on the Rhizosphere Fungal Community of Allium fistulosum L.

  • Chunxiang Ji;Yingyue Li;Qingchen Xiao;Zishan Li;Boyan Wang;Xiaowan Geng;Keqing Lin;Qing Zhang;Yuan Jin;Yuqian Zhai;Xiaoyu Li;Jin Chen
    • Journal of Microbiology and Biotechnology
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    • 제33권8호
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    • pp.1013-1022
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    • 2023
  • Arbuscular mycorrhizal fungi (AMF) are widespread soil endophytic fungi, forming mutualistic relationships with the vast majority of land plants. Biochar (BC) has been reported to improve soil fertility and promote plant growth. However, limited studies are available concerning the combined effects of AMF and BC on soil community structure and plant growth. In this work, a pot experiment was designed to investigate the effects of AMF and BC on the rhizosphere microbial community of Allium fistulosum L. Using Illumina high-throughput sequencing, we showed that inoculation of AMF and BC had a significant impact on soil microbial community composition, diversity, and versatility. Increases were observed in both plant growth (the plant height by 8.6%, shoot fresh weight by 12.1%) and root morphological traits (average diameter by 20.5%). The phylogenetic tree also showed differences in the fungal community composition in A. fistulosum. In addition, Linear discriminant analysis (LDA) effect size (LEfSe) analysis revealed that 16 biomarkers were detected in the control (CK) and AMF treatment, while only 3 were detected in the AMF + BC treatment. Molecular ecological network analysis showed that the AMF + BC treatment group had a more complex network of fungal communities, as evidenced by higher average connectivity. The functional composition spectrum showed significant differences in the functional distribution of soil microbial communities among different fungal genera. The structural equation model (SEM) confirmed that AMF could improve the microbial multifunctionality by regulating the rhizosphere fungal diversity and soil properties. Our findings provide new information on the effects of AMF and biochar on plants and soil microbial communities.

Whole genome sequence of Staphylococcus aureus strain RMI-014804 isolated from pulmonary patient sputum via next-generation sequencing technology

  • Ayesha, Wisal;Asad Ullah;Waheed Anwar;Carlos M. Morel;Syed Shah Hassan
    • Genomics & Informatics
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    • 제21권3호
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    • pp.34.1-34.10
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    • 2023
  • Nosocomial infections, commonly referred to as healthcare-associated infections, are illnesses that patients get while hospitalized and are typically either not yet manifest or may develop. One of the most prevalent nosocomial diseases in hospitalized patients is pneumonia, among the leading causes of mortality and morbidity. Viral, bacterial, and fungal pathogens cause pneumonia. More severe introductions commonly included Staphylococcus aureus, which is at the top of bacterial infections, per World Health Organization reports. The staphylococci, S. aureus, strain RMI-014804, mesophile, on-sporulating, and non-motile bacterium, was isolated from the sputum of a pulmonary patient in Pakistan. Many characteristics of S. aureus strain RMI-014804 have been revealed in this paper, with complete genome sequence and annotation. Our findings indicate that the genome is a single circular 2.82 Mbp long genome with 1,962 protein-coding genes, 15 rRNA, 49 tRNA, 62 pseudogenes, and a GC content of 28.76%. As a result of this genome sequencing analysis, researchers will fully understand the genetic and molecular basis of the virulence of the S. aureus bacteria, which could help prevent the spread of nosocomial infections like pneumonia. Genome analysis of this strain was necessary to identify the specific genes and molecular mechanisms that contribute to its pathogenicity, antibiotic resistance, and genetic diversity, allowing for a more in-depth investigation of its pathogenesis to develop new treatments and preventive measures against infections caused by this bacterium.

Ginsenoside Rk1 inhibits HeLa cell proliferation through an endoplasmic reticulum signaling pathway

  • Qiuyang Li;Hang Sun;Shiwei Liu;Jinxin Tang;Shengnan Liu;Pei Yin;Qianwen Mi;Jingsheng Liu;Lei yu;Yunfeng Bi
    • Journal of Ginseng Research
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    • 제47권5호
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    • pp.645-653
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    • 2023
  • Background: Changes to work-life balance has increased the incidence of cervical cancer among younger people. A minor ginseng saponin known as ginsenoside Rk1 can inhibit the growth and survival of human cancer cells; however, whether ginsenoside Rk1 inhibits HeLa cell proliferation is unknown. Methods and results: Ginsenoside Rk1 blocked HeLa cells in the G0/G1 phase in a dose-dependent manner and inhibited cell division and proliferation. Ginsenoside Rk1 markedly also activated the apoptotic signaling pathway via caspase 3, PARP, and caspase 6. In addition, ginsenoside Rk1 increased LC3B protein expression, indicating the promotion of the autophagy signaling pathway. Protein processing in the endoplasmic reticulum signaling pathway was downregulated in Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses, consistent with teal-time quantitative PCR and western blotting that showed YOD1, HSPA4L, DNAJC3, and HSP90AA1 expression levels were dramatically decreased in HeLa cells treated with ginsenoside Rk1, with YOD1 was the most significantly inhibited by ginsenoside Rk1 treatment. Conclusion: These findings indicate that the toxicity of ginsenoside Rk1 in HeLa cells can be explained by the inhibition of protein synthesis in the endoplasmic reticulum and enhanced apoptosis, with YOD1 acting as a potential target for cervical cancer treatment.

유용한 바실러스의 토양 접종에 따른 토착 세균 군집의 변화 (Changes in Resident Soil Bacterial Communities in Response to Inoculation of Soil with Beneficial Bacillus spp.)

  • 김이슬;김상윤;안주희;상미경;원항연;송재경
    • 한국미생물·생명공학회지
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    • 제46권3호
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    • pp.253-260
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    • 2018
  • 유용미생물은 임업과 축산 분야에 활용될 뿐만 아니라 병해충 방제와 작물 생육 증진 등의 용도로 농업에서 널리 이용되고 있다. 하지만 유용미생물의 토양에서의 생존율과 정착율에 대한 연구는 미미한 형편이다. 본 연구에서는 마이크로코즘을 이용해 바실러스 3 균주를 토양에 처리한 후, 이들의 토양 내 생존능을 정량 PCR을 이용하여 13일 동안 정량적으로 분석하였다. 또한 Illumina MiSeq 플랫폼을 이용하여 바실러스 3 균주 처리구와 대조구의 토양미생물 군집 분포를 비교 및 분석하였다. 바실러스 3 균주의 처리 직후 토양 내 밀도는 건조토양 1 그람당 평균 $4.4{\times}10^6$ 유전자수로 대조구에 비해 1,000배 이상 높았다. 바실러스 균주의 토양 내 밀도는 처리 후 약 일주일 간 유지되었고 그 후부터는 유의성 있게 감소하였지만 여전히 대조구보다 100배 이상 높았다. 바실러스 균주 처리 후 토양 내 미생물 군집 구조 분석 결과, 대조구와 처리구 모두 Acidobacteria 문($26.3{\pm}0.9%$), Proteobacteria 문($24.2{\pm}0.5%$), Chloroflexi 문($11.1{\pm}0.4%$), Actinobacteria 문($9.7{\pm}2.5%$)에 속하는 세균이 우점하였다. 대조구 대비 처리구에서 Actinobacteria 문의 비율은 뚜렷하게 감소하였지만 Bacteroidetes 문과 Firmicutes 문의 비율은 증가하는 경향이었다. 속 수준에서 바실러스 3 균주를 처리함에 따라 일부 세균 군집의 종 풍부도를 변화되었고, 결국 전체 토착 미생물 군집 구조가 변화되었음을 확인할 수 있었다. 본 연구에서 수행한 유용한 바실러스의 토양 접종 후 이들의 토양 내 생존능 분석 및 토착 세균 군집의 변화는 유용미생물을 생물적 제제로 시설재배지에 사용할 때 중요한 정보를 제공할 것으로 판단된다.