• Title/Summary/Keyword: High-throughput Sequencing

Search Result 220, Processing Time 0.029 seconds

Fast and Precise: How to Measure Meiotic Crossovers in Arabidopsis

  • Kim, Heejin;Choi, Kyuha
    • Molecules and Cells
    • /
    • v.45 no.5
    • /
    • pp.273-283
    • /
    • 2022
  • During meiosis, homologous chromosomes (homologs) pair and undergo genetic recombination via assembly and disassembly of the synaptonemal complex. Meiotic recombination is initiated by excess formation of DNA double-strand breaks (DSBs), among which a subset are repaired by reciprocal genetic exchange, called crossovers (COs). COs generate genetic variations across generations, profoundly affecting genetic diversity and breeding. At least one CO between homologs is essential for the first meiotic chromosome segregation, but generally only one and fewer than three inter-homolog COs occur in plants. CO frequency and distribution are biased along chromosomes, suppressed in centromeres, and controlled by pro-CO, anti-CO, and epigenetic factors. Accurate and high-throughput detection of COs is important for our understanding of CO formation and chromosome behavior. Here, we review advanced approaches that enable precise measurement of the location, frequency, and genomic landscapes of COs in plants, with a focus on Arabidopsis thaliana.

First Report of Tomato Spotted Wilt Virus in Oxypetalum coeruleum in Korea (옥시페탈룸에서 발생한 토마토반점위조바이러스 국내 첫 보고)

  • Eseul, Baek;Peter, Palukaitis;Ju-Yeon, Yoon
    • Research in Plant Disease
    • /
    • v.28 no.4
    • /
    • pp.231-236
    • /
    • 2022
  • Oxypetalum coeruleum, commonly known as Tweedia, is a perennial herbaceous plant of the Apocynaceae family native to southern Brazil and Uruguay. Tweedia plants are grown as one of the most popular ornamental flowers for floral arrangement in Korea. In May 2021, several tweedia plants in a single greenhouse in Gimje, Jeollabuk-do were found to show virus-like symptoms including necrotic rings, vein-clearing, chlorotic mottle, and mosaic on the leaves, and necrosis on the stems. Here, we have identified tomato spotted wilt virus (TSWV) in symptomatic tweedia leaves by applying high-throughput RNA sequencing. In the result, a single infection by TSWV was verified without mixed infections of different virus species. To confirm the presence of TSWV, a reverse transcription polymerase chain reaction was performed with a specific primer set to the N gene of TSWV. The complete genomic sequence of L, M, and S segments of TSWV 'Oxy' isolate were determined and deposited in GenBank under accession numbers LC671525, LC671638, and LC671639, respectively. In the phylogenetic tree analysis by maximum likelihood method, 'Oxy' isolate showed a high relationship with TSWV 'Gumi' isolate from Gerbera jamesonii in Gyeongsangbuk-do, Korea; for all three RNA segments. To our knowledge, this is the first report of TSWV infection of O. coeruleum in Korea.

A combined approach to evaluate activity and structure of soil microbial community in long-term heavy metals contaminated soils

  • Wang, Tianqi;Yuan, Zhimin;Yao, Jun
    • Environmental Engineering Research
    • /
    • v.23 no.1
    • /
    • pp.62-69
    • /
    • 2018
  • In the present study, long-term heavy metals (HMs) contaminated soil samples from a well-known Pb/Zn smelting area in the southwest of China were collected, and physicochemical and biological characteristics of these samples were evaluated. Soil samples contained different concentrations of HMs, namely Pb, Zn, Cu, and Cd. Enzyme activity analyses combined with microcalorimetric analysis were used for soil microbial activity evaluation. Results showed that two soil samples, containing almost the highest concentrations of HMs, also shared the greatest microbial activities. Based on correlation coefficient analysis, high microbial activity in heavily HMs contaminated soil might be due to the high contents of soil organic matter and available phosphorus in these samples. High-throughput sequencing technique was used for microbial community structure analysis. High abundance of genera Sphingomonas and Thiobacillus were also observed in these two heavily contaminated soils, suggesting that bacteria belonging to these two genera might be further isolated from these contaminated soils and applied for future studies of HMs remediation. Results of present study would contribute to the evaluation of microbial communities and isolation of microbial resources to remediate HMs pollution.

Information Technology Infrastructure for Agriculture Genotyping Studies

  • Pardamean, Bens;Baurley, James W.;Perbangsa, Anzaludin S.;Utami, Dwinita;Rijzaani, Habib;Satyawan, Dani
    • Journal of Information Processing Systems
    • /
    • v.14 no.3
    • /
    • pp.655-665
    • /
    • 2018
  • In efforts to increase its agricultural productivity, the Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development has conducted a variety of genomic studies using high-throughput DNA genotyping and sequencing. The large quantity of data (big data) produced by these biotechnologies require high performance data management system to store, backup, and secure data. Additionally, these genetic studies are computationally demanding, requiring high performance processors and memory for data processing and analysis. Reliable network connectivity with large bandwidth to transfer data is essential as well as database applications and statistical tools that include cleaning, quality control, querying based on specific criteria, and exporting to various formats that are important for generating high yield varieties of crops and improving future agricultural strategies. This manuscript presents a reliable, secure, and scalable information technology infrastructure tailored to Indonesian agriculture genotyping studies.

Transferability of Cupped Oyster EST (Expressed Sequence Tag)-Derived SNP (Single Nucleotide Polymorphism) Markers to Related Crassostrea and Ostrea Species

  • Kim, Woo-Jin;Jung, Hyungtaek;Shin, Eun-Ha;Baek, Ilseon
    • The Korean Journal of Malacology
    • /
    • v.30 no.3
    • /
    • pp.197-210
    • /
    • 2014
  • Single nucleotide polymorphisms (SNPs) are widely acknowledged as the marker of choice for many genetic and genomic applications because they show co-dominant inheritance, are highly abundant across genomes and are suitable for high-throughput genotyping. Here we evaluated the applicability of SNP markers developed from Crassostrea gigas and C. virginica expressed sequence tags (ESTs) in closely related Crassostrea and Ostrea species. A total of 213 putative interspecific level SNPs were identified from re-sequencing data in six amplicons, yielding on average of one interspecific level SNP per seven bp. High polymorphism levels were observed and the high success rate of transferability show that genic EST-derived SNP markers provide an efficient method for rapid marker development and SNP discovery in closely related oyster species. The six EST-SNP markers identified here will provide useful molecular tools for addressing questions in molecular ecology and evolution studies including for stock analysis (pedigree monitoring) in related oyster taxa.

Pyrosequencing Based Detection of Rifampicin or Isoniazid Resistant in Mycobacterium tuberculosis (Pyrosequencing 분석법을 이용한 Rifampicin과 Isoniazid 결핵약제내성의 빠른 검사법)

  • Oh, Seo-Young;Kim, Hyo-Bin;Shin, Min-Sik;Kim, Jin-Wook;Park, Sung-Hwuy
    • Korean Journal of Clinical Laboratory Science
    • /
    • v.41 no.1
    • /
    • pp.24-30
    • /
    • 2009
  • Rifampicin (RIF) and isoniazid (INH) are the most important drug for the treatment of Mycobacterium tuberculosis. Mutations correlated to rifampicin and isoniazid-resistance have been detected in rpoB gene and katG gene, respectively. Of the rifampicin-resistant isolates, 90% showed mutations in rpoB gene at codon 507 to 533. Isoniazid-resistant isolates analysed had a mutation in katG at codon 315. The aim of this study is to develop a pyrosequencing-based approach for rapid detection of ripampin or isoniazid resistant M. tuberculosis based on characterization of all possible mutation in the target region. For this study, the DNA selected from 35 cases of MTB PCR positive clinical sample such as bronchial washing, sputum, and pleural fluid. RIF or INH resistant was analyzed by pyrosequencing data of rpoB and katG gene. 28 (80%) and 7 (20%) of 35 MTB PCR positive DNAs were occured rifampicin-sensitivity and resistant, respectively. For INH, 30 (85.7%) and 5 (14.5%) cases were detected isoniazid-sensitivity and resistant, respectively. When pyrosequencing analysis was compared with ABI sequencing analysis, both analysis were presented same result, but pyrosequencing analysis was more rapid than ABI sequencing analysis. In conclusion, we found that pyrosequencing technology offers high accuracy, specificity, short turn around time and a high throughput in detection of rifampicin or isoniazid resistance in M. tuberculosis.

  • PDF

Characterization of Vaginal Microbiota Associated with Pregnancy Outcomes of Artificial Insemination in Dairy Cows

  • Chen, Shi-Yi;Deng, Feilong;Zhang, Ming;Jia, Xianbo;Lai, Song-Jia
    • Journal of Microbiology and Biotechnology
    • /
    • v.30 no.6
    • /
    • pp.804-810
    • /
    • 2020
  • The profitability of the dairy and beef industries is largely affected by the actually achieved reproductive efficiency. Although a large proportion of cows worldwide are bred by artificial insemination (AI) services, many potential factors affecting the outcome of pregnancy by AI remain to be addressed. In the present study, we investigated the vaginal microbiota by high-throughput sequencing of 16S rRNA gene and analyzed their association with differential pregnancy outcomes (i.e., pregnant vs. nonpregnant) of multiple AI services in dairy cows. Sequencing of the V3-V4 region totally produced 512,046 high-quality sequences that were computationally clustered into 2,584 operational taxonomic units (OTUs). All OTUs were taxonomically assigned to 10 bacterial phyla. There were statistically significant differences among the three AI service times (T1, T2 and T3) with respect to the Shannon index and number of observed OTUs (p < 0.05). Bray-Curtis distance-based PCoA analysis also revealed that T2 group could be significantly distinguished from T1 and T3. However, no significant difference between the pregnant and nonpregnant cows was found in confidence regarding both alpha diversity and beta diversity. These results could help us better understand the possible influence of vaginal microbial community on pregnancy outcomes of AI service in cows.

Single Nucleotide Polymorphism Marker Discovery from Transcriptome Sequencing for Marker-assisted Backcrossing in Capsicum

  • Kang, Jin-Ho;Yang, Hee-Bum;Jeong, Hyeon-Seok;Choe, Phillip;Kwon, Jin-Kyung;Kang, Byoung-Cheorl
    • Horticultural Science & Technology
    • /
    • v.32 no.4
    • /
    • pp.535-543
    • /
    • 2014
  • Backcross breeding is the method most commonly used to introgress new traits into elite lines. Conventional backcross breeding requires at least 4-5 generations to recover the genomic background of the recurrent parent. Marker-assisted backcrossing (MABC) represents a new breeding approach that can substantially reduce breeding time and cost. For successful MABC, highly polymorphic markers with known positions in each chromosome are essential. Single nucleotide polymorphism (SNP) markers have many advantages over other marker systems for MABC due to their high abundance and amenability to genotyping automation. To facilitate MABC in hot pepper (Capsicum annuum), we utilized expressed sequence tags (ESTs) to develop SNP markers in this study. For SNP identification, we used Bukang $F_1$-hybrid pepper ESTs to prepare a reference sequence through de novo assembly. We performed large-scale transcriptome sequencing of eight accessions using the Illumina Genome Analyzer (IGA) IIx platform by Solexa, which generated small sequence fragments of about 90-100 bp. By aligning each contig to the reference sequence, 58,151 SNPs were identified. After filtering for polymorphism, segregation ratio, and lack of proximity to other SNPS or exon/intron boundaries, a total of 1,910 putative SNPs were chosen and positioned to a pepper linkage map. We further selected 412 SNPs evenly distributed on each chromosome and primers were designed for high throughput SNP assays and tested using a genetic diversity panel of 27 Capsicum accessions. The SNP markers clearly distinguished each accession. These results suggest that the SNP marker set developed in this study will be valuable for MABC, genetic mapping, and comparative genome analysis.

The impact of diet on the composition and relative abundance of rumen microbes in goat

  • Liu, Kaizhen;Xu, Qin;Wang, Lizhi;Wang, Jiwen;Guo, Wei;Zhou, Meili
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.30 no.4
    • /
    • pp.531-537
    • /
    • 2017
  • Objective: This experiment was conducted to explore the impact of diet on the ruminal microbial community in goats. Methods: Twelve goats were divided into two groups and fed complete feed (CF) or all forage (AF) diet. The total microbial DNAs in the rumen liquid were extracted. The V4 region of microbial 16S rRNA genes was amplified and sequenced using high-throughput. Information of sequences was mainly analyzed by QIIME 1.8.0. Results: The results showed that Bacteroidetes and Firmicutes were the most predominant microbial phyla in the rumen of all goats. At genus level, the abundance of fiber-digesting bacteria such as Ruminococcus and Lachnospiracea incertae sedis was significantly higher in AF than that in CF, while the levels of fat-degrading bacterium Anaerovibrio and protein-degrading bacterium Pseudomonas were opposite. The core shared genera, Prevotella and Butyrivibrio were widespread in the rumen of goats and no significant difference was observed in relative abundance between groups. Conclusion: We concluded that the richness of fiber-, protein-, and fat-digesting bacteria was affected by diet and tended to increase with the rise of their corresponding substrate contents in the ration; some bacteria shared by all goats maintained stable despite the difference in the ration, and they might be essential in maintaining the normal function of rumen.

High-throughput Gene Expression Analysis to Investigate Host-pathogen Interaction in Avian Coccidiosis

  • Lillehoj Hyun, S.
    • Korean Journal of Poultry Science
    • /
    • v.34 no.1
    • /
    • pp.77-83
    • /
    • 2007
  • Poultry products including meat and eggs constitute a major protein source in the American diet and disease-causing pathogens represent major challenges to the poultry industry. More than 95% of pathogens enter the host through the mucosal surfaces of the respiratory, digestive and reproductive tracts and over the past few decades, the two main mechanisms used to control diseases have been the use of vaccines and antibiotics. However, in the poultry industry, there are mounting concerns over the ability of current vaccines to adequately protect against emerging hyper-virulent strains of pathogens and a lack of suitable, cost effective adjuvants. Thorough investigation of the immunogenetic responses involved in host-pathogen interactions will lead to the development of new and effective strategies for improving poultry health, food safety and the economic viability of the US poultry industry. In this paper, I describe the development of immunogenomic and proteomic tools to fundamentally determine and characterize the immunological mechanisms of the avian host to economically significant mucosal pathogens such as Eimeria. Recent completion of poultry genome sequencing and the development of several tissue-specific cDNA libraries in chickens are facilitating the rapid application of functional immunogenomics in the poultry disease research. Furthermore, research involving functional genomics, immunology and bioinformatics is providing novel insights into the processes of disease and immunity to microbial pathogens at mucosal surfaces. In this presentation, a new strategy of global gene expression using avian macrophage (AMM) to characterize the multiple pathways related to the variable immune responses of the host to Eimeria is described. This functional immunogenomics approach will increase current understanding of how mucosal immunity to infectious agents operates, and how it may be enhanced to enable the rational development of new and effective strategies against coccidiosis and other mucosal pathogens.