scholarly journals Application of high-throughput amplicon sequencing-based SSR genotyping in genetic background screening

BMC Genomics ◽  
2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Tiantian Li ◽  
Zhiwei Fang ◽  
Hai Peng ◽  
Junfei Zhou ◽  
Pengcheng Liu ◽  
...  
Keyword(s):  
High Throughput ◽  
Genetic Background ◽  
Amplicon Sequencing ◽  
Ssr Genotyping

scholarly journals Extremely Halophilic Biohydrogen Producing Microbial Communities from High-Salinity Soil and Salt Evaporation Pond

Fuels ◽  
2021 ◽  
Vol 2 (2) ◽  
pp. 241-252
Author(s):  
Dyah Asri Handayani Taroepratjeka ◽  
Tsuyoshi Imai ◽  
Prapaipid Chairattanamanokorn ◽  
Alissara Reungsang
Keyword(s):  
Microbial Communities ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
High Salinity ◽  
Amplicon Sequencing ◽  
Spatial Proximity ◽  
Lignocellulosic Waste ◽  
Evaporation Pond ◽  
Operational Taxonomic Units ◽  
Determining Factor

Extreme halophiles offer the advantage to save on the costs of sterilization and water for biohydrogen production from lignocellulosic waste after the pretreatment process with their ability to withstand extreme salt concentrations. This study identifies the dominant hydrogen-producing genera and species among the acclimatized, extremely halotolerant microbial communities taken from two salt-damaged soil locations in Khon Kaen and one location from the salt evaporation pond in Samut Sakhon, Thailand. The microbial communities’ V3–V4 regions of 16srRNA were analyzed using high-throughput amplicon sequencing. A total of 345 operational taxonomic units were obtained and the high-throughput sequencing confirmed that Firmicutes was the dominant phyla of the three communities. Halanaerobium fermentans and Halanaerobacter lacunarum were the dominant hydrogen-producing species of the communities. Spatial proximity was not found to be a determining factor for similarities between these extremely halophilic microbial communities. Through the study of the microbial communities, strategies can be developed to increase biohydrogen molar yield.

scholarly journals Different Amplicon Targets for Sequencing-Based Studies of Fungal Diversity

2017 ◽  
Vol 83 (17) ◽  
Author(s):  
Francesca De Filippis ◽  
Manolo Laiola ◽  
Giuseppe Blaiotta ◽  
Danilo Ercolini
Keyword(s):  
Microbial Ecology ◽  
High Throughput ◽  
Biological Samples ◽  
Fungal Diversity ◽  
High Throughput Sequencing ◽  
Amplicon Sequencing ◽  
Fungal Species ◽  
Fungal Communities ◽  
Its2 Region ◽  
Mock Community

ABSTRACT Target-gene amplicon sequencing is the most exploited high-throughput sequencing application in microbial ecology. The targets are taxonomically relevant genes, with 16S rRNA being the gold standard for bacteria. As for fungi, the most commonly used target is the internal transcribed spacer (ITS). However, the uneven ITS length among species may promote preferential amplification and sequencing and incorrect estimation of their abundance. Therefore, the use of different targets is desirable. We evaluated the use of three different target amplicons for the characterization of fungal diversity. After an in silico primer evaluation, we compared three amplicons (the ITS1-ITS2 region [ITS1-2], 18S ribosomal small subunit RNA, and the D1/D2 domain of the 26S ribosomal large subunit RNA), using biological samples and a mock community of common fungal species. All three targets allowed for accurate identification of the species present. Nevertheless, high heterogeneity in ITS1-2 length was found, and this caused an overestimation of the abundance of species with a shorter ITS, while both 18S and 26S amplicons allowed for more reliable quantification. We demonstrated that ITS1-2 amplicon sequencing, although widely used, may lead to an incorrect evaluation of fungal communities, and efforts should be made to promote the use of different targets in sequencing-based microbial ecology studies. IMPORTANCE Amplicon-sequencing approaches for fungi may rely on different targets affecting the diversity and abundance of the fungal species. An increasing number of studies will address fungal diversity by high-throughput amplicon sequencing. The description of the communities must be accurate and reliable in order to draw useful insights and to address both ecological and biological questions. By analyzing a mock community and several biological samples, we demonstrate that using different amplicon targets may change the results of fungal microbiota analysis, and we highlight how a careful choice of the target is fundamental for a thorough description of the fungal communities.

scholarly journals Characterization of Holstein and Normande whole milk miRNomes highlights breed specificities

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
S. Le Guillou ◽  
A. Leduc ◽  
J. Laubier ◽  
S. Barbey ◽  
M.-N. Rossignol ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Milk Production ◽  
High Throughput ◽  
Genetic Background ◽  
Molecular Mechanisms ◽  
High Throughput Sequencing ◽  
Whole Milk ◽  
Mammary Morphogenesis ◽  
Regulatory Properties

AbstractThe concept of milk as a healthy food has opened the way for studies on milk components, from nutrients to microRNAs, molecules with broad regulatory properties present in large quantities in milk. Characterization of these components has been performed in several species, such as humans and bovine, depending on the stages of lactation. Here, we have studied the variation in milk microRNA composition according to genetic background. Using high throughput sequencing, we have characterized and compared the milk miRNomes of Holstein and Normande cattle, dairy breeds with distinct milk production features, in order to highlight microRNAs that are essential for regulation of the lactation process. In Holstein and Normande milk, 2,038 and 2,030 microRNAs were identified, respectively, with 1,771 common microRNAs, of which 1,049 were annotated and 722 were predicted. The comparison of the milk miRNomes of two breeds allowed to highlight 182 microRNAs displaying significant differences in the abundance. They are involved in the regulation of lipid metabolism and mammary morphogenesis and development, which affects lactation. Our results provide new insights into the regulation of molecular mechanisms involved in milk production.

scholarly journals Application of a High-Throughput Amplicon Sequencing Method to Chart the Bacterial Communities that Are Associated with European Fermented Meats from Different Origins

Foods ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 1247
Author(s):  
Emiel Van Reckem ◽  
Christina Charmpi ◽  
David Van der Veken ◽  
Wim Borremans ◽  
Luc De Vuyst ◽  
...  
Keyword(s):  
Microbial Communities ◽  
High Throughput ◽  
Bacterial Communities ◽  
Geographical Origin ◽  
Salt Content ◽  
Amplicon Sequencing ◽  
Taxonomic Resolution ◽  
Future Application ◽  
Processing Conditions ◽  
The Impact

Insight into the microbial species diversity of fermented meats is not only paramount to gain control over quality development, but also to better understand the link with processing technology and geographical origin. To study the composition of the microbial communities, the use of culture-independent methods is increasingly popular but often still suffers from drawbacks, such as a limited taxonomic resolution. This study aimed to apply a previously developed high-throughput amplicon sequencing (HTS) method targeting the 16S rRNA and tuf genes to characterize the bacterial communities in European fermented meats in greater detail. The data obtained broadened the view on the microbial communities that were associated with the various products examined, revealing the presence of previously underreported subdominant species. Moreover, the composition of these communities could be linked to the specificities of individual products, in particular pH, salt content, and geographical origin. In contrast, no clear links were found between the volatile organic compound profiles of the different products and the country of origin, distinct processing conditions, or microbial communities. Future application of the HTS method offers the potential to further unravel complex microbial communities in fermented meats, as well as to assess the impact of different processing conditions on microbial consortia.

scholarly journals Correction for Gaby et al., “Diazotroph Community Characterization via a High-Throughput nifH Amplicon Sequencing and Analysis Pipeline”

2019 ◽  
Vol 85 (11) ◽  
Author(s):  
John Christian Gaby ◽  
Lavanya Rishishwar ◽  
Lina C. Valderrama-Aguirre ◽  
Stefan J. Green ◽  
Augusto Valderrama-Aguirre ◽  
...  
Keyword(s):  
High Throughput ◽  
Amplicon Sequencing ◽  
Analysis Pipeline

Using PacBio Long-Read High-Throughput Microbial Gene Amplicon Sequencing To Evaluate Infant Formula Safety

2016 ◽  
Vol 64 (37) ◽  
pp. 6993-7001 ◽  
Author(s):  
Yi Zheng ◽  
Xiaoxia Xi ◽  
Haiyan Xu ◽  
Qiangchuan Hou ◽  
Yanfei Bian ◽  
...  
Keyword(s):  
Infant Formula ◽  
High Throughput ◽  
Amplicon Sequencing ◽  
Long Read

scholarly journals An improved method for utilizing high‐throughput amplicon sequencing to determine the diets of insectivorous animals

2019 ◽  
Vol 19 (1) ◽  
pp. 176-190 ◽  
Author(s):  
Michelle A. Jusino ◽  
Mark T. Banik ◽  
Jonathan M. Palmer ◽  
Amy K. Wray ◽  
Lei Xiao ◽  
...  
Keyword(s):  
High Throughput ◽  
Amplicon Sequencing ◽  
Improved Method

scholarly journals High-throughput amplicon sequencing of the full-length 16S rRNA gene with single-nucleotide resolution

2019 ◽  
Vol 47 (18) ◽  
pp. e103-e103 ◽  
Author(s):  
Benjamin J Callahan ◽  
Joan Wong ◽  
Cheryl Heiner ◽  
Steve Oh ◽  
Casey M Theriot ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
High Throughput ◽  
Amplicon Sequencing ◽  
Full Length ◽  
Rrna Gene ◽  
Single Nucleotide ◽  
Full Complement ◽  
Nucleotide Resolution ◽  
Single Nucleotide Resolution

AbstractTargeted PCR amplification and high-throughput sequencing (amplicon sequencing) of 16S rRNA gene fragments is widely used to profile microbial communities. New long-read sequencing technologies can sequence the entire 16S rRNA gene, but higher error rates have limited their attractiveness when accuracy is important. Here we present a high-throughput amplicon sequencing methodology based on PacBio circular consensus sequencing and the DADA2 sample inference method that measures the full-length 16S rRNA gene with single-nucleotide resolution and a near-zero error rate. In two artificial communities of known composition, our method recovered the full complement of full-length 16S sequence variants from expected community members without residual errors. The measured abundances of intra-genomic sequence variants were in the integral ratios expected from the genuine allelic variants within a genome. The full-length 16S gene sequences recovered by our approach allowed Escherichia coli strains to be correctly classified to the O157:H7 and K12 sub-species clades. In human fecal samples, our method showed strong technical replication and was able to recover the full complement of 16S rRNA alleles in several E. coli strains. There are likely many applications beyond microbial profiling for which high-throughput amplicon sequencing of complete genes with single-nucleotide resolution will be of use.

scholarly journals The Grapevine and Wine Microbiome: Insights from High-Throughput Amplicon Sequencing

2017 ◽  
Vol 8 ◽  
Author(s):  
Horatio H. Morgan ◽  
Maret du Toit ◽  
Mathabatha E. Setati
Keyword(s):  
High Throughput ◽  
Amplicon Sequencing
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