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Zhang, Qiuting

Publications and source records attributed to Zhang, Qiuting.

7 records
All sourcesNASA NTRSDOE OSTI
DOE OSTI2023

Effects of error, chimera, bias, and GC content on the accuracy of amplicon sequencing

ABSTRACT Targeted amplicon sequencing is widely used in microbial ecology studies. However, sequencing artifacts and amplification biases are of great concern. To identify sources of these artifacts, a systematic analysis was performed using mock communities comprised of 16S rRNA genes from 33 bacterial strains. Our results indicated that while sequencing errors were generally isolated to low-abundance operational taxonomic units, chimeric sequences were a major source of artifacts. Singleton and doubleton sequences were primarily chimeras. Formation of chimeric sequences was significantly correlated with the GC content of the targeted sequences. Low-GC-content mock community members exhibited lower rates of chimeric sequence formation. GC content also had a large impact on sequence recovery. The quantitative capacity was notably limited, with substantial recovery variations and weak correlation between anticipated and observed strain abundances. The mock community strains with higher GC content had higher recovery rates than strains with lower GC content. Amplification bias was also observed due to the differences in primer affinity. A two-step PCR strategy reduced the number of chimeric sequences by half. In addition, comparative analyses based on the mock communities showed that several widely used sequence processing pipelines/methods, including DADA2, Deblur, UCLUST, UNOISE, and UPARSE, had different advantages and disadvantages in artifact removal and rare species detection. These results are important for improving sequencing quality and reliability and developing new algorithms to process targeted amplicon sequences. IMPORTANCE Amplicon sequencing of targeted genes is the predominant approach to estimate the membership and structure of microbial communities. However, accurate reconstruction of community composition is difficult due to sequencing errors, and other methodological biases and effective approaches to overcome these challenges are essential. Using a mock community of 33 phylogenetically diverse strains, this study evaluated the effect of GC content on sequencing results and tested different approaches to improve overall sequencing accuracy while characterizing the pros and cons of popular amplicon sequence data processing approaches. The sequencing results from this study can serve as a benchmarking data set for future algorithmic improvements. Furthermore, the new insights on sequencing error, chimera formation, and GC bias from this study will help enhance the quality of amplicon sequencing studies and support the development of new data analysis approaches.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Microbial Functional Responses Explain Alpine Soil Carbon Fluxes under Future Climate Scenarios

Soil microorganisms are sensitive to temperature in cold ecosystems, but it remains unclear how microbial responses are modulated by other important climate drivers, such as precipitation changes. Here, we examine the effects of six in situ warming and/or precipitation treatments in alpine grasslands on microbial communities, plants, and soil carbon fluxes. These treatments differentially affected soil carbon fluxes, gross primary production, and microbial communities. Variations of soil CO 2 and CH 4 fluxes across all sites significantly (r > 0.70, P < 0.050) correlated with relevant microbial functional abundances but not bacterial or fungal abundances. Given tight linkages between microbial functional traits and ecosystem functionality, we conclude that future soil carbon fluxes in alpine grasslands can be predicted by microbial carbon-degrading capacities.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2019

Author Correction: Global diversity and biogeography of bacterial communities in wastewater treatment plants

An amendment to this paper has been published and can be accessed via a link at the top of the paper. Correction to: Nature Microbiology https://doi.org/10.1038/s41564-019-0426-5, published online 13 May 2019. In the version of this Article originally published, the name of the author ‘Mathew Robert Brown’ was incorrectly written as ‘Mathew Brown’ in the main author list and as ‘Matthew Brown’ in the Global Water Microbiome Consortium list. In addition, in the Global Water Microbiome Consortium list, the names of the authors ‘Kevin F. Boehnke’, ‘Janeth Sanabria’ and ‘Adalberto Noyola’ were incorrectly written as ‘Kevin Boehnke’, ‘Janeth Sanabria Gómez’ and ‘Adalberto Noyola Robles’, respectively. The names have now been corrected and the author initials in the author contributions section updated accordingly.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2019

Global diversity and biogeography of bacterial communities in wastewater treatment plants

Microorganisms in wastewater treatment plants (WWTPs) are essential for water purification to protect public and environmental health. However, the diversity of microorganisms and the factors that control it are poorly understood. Using a systematic global-sampling effort, we analysed the 16S ribosomal RNA gene sequences from ~1,200 activated sludge samples taken from 269 WWTPs in 23 countries on 6 continents. Our analyses revealed that the global activated sludge bacterial communities contain ~1 billion bacterial phylotypes with a Poisson lognormal diversity distribution. Despite this high diversity, activated sludge has a small, global core bacterial community ( n = 28 operational taxonomic units) that is strongly linked to activated sludge performance. Meta-analyses with global datasets associate the activated sludge microbiomes most closely to freshwater populations. In contrast to macroorganism diversity, activated sludge bacterial communities show no latitudinal gradient. Furthermore, their spatial turnover is scale-dependent and appears to be largely driven by stochastic processes (dispersal and drift), although deterministic factors (temperature and organic input) are also important. Finally, our findings enhance our mechanistic understanding of the global diversity and biogeography of activated sludge bacterial communities within a theoretical ecology framework and have important implications for microbial ecology and wastewater treatment processes.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2019

High variations of methanogenic microorganisms drive full-scale anaerobic digestion process

Anaerobic digestion is one of the most successful waste management strategies worldwide, wherein microorganisms play an essential role in reducing organic pollutants and producing renewable energy. However, variations of microbial community in full-scale anaerobic digesters, particularly functional groups relevant to biogas production, remain elusive. Here, we examined microbial community in a year-long monthly time series of 3 full-scale anaerobic digesters. We observed substantial diversification in community composition, with only a few abundant OTUs (e.g. Clostridiales, Anaerolineaceae and Methanosaeta) persistently present across different samples. Similarly, there were high variations in relative abundance of methanogenic archaea and methanogenic genes, which were positively correlated (r 2 = 0.530, P < 0.00 1). Variations of methanogens explained 55.7% of biogas producing rates, much higher than the explanatory percentage of environmental parameters (16.4%). Hydrogenotrophic methanogens, especially abundant Methanomicrobiales taxa, were correlated with biogas production performance (r = 0.665, P < 0.001) and nearly all methanogenic genes (0.430 < r < 0.735, P < 0.012). Given that methanogenic archaea or genes are well established for methanogenesis, we conclude that high variations in methanogenic traits (e.g. taxa or genes) are responsible for biogas production variations in full-scale anaerobic digesters.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2019

Microbial functional traits are sensitive indicators of mild disturbance by lamb grazing

Mild disturbances are prevalent in the environment, which may not be easily notable but could have considerable ecological consequences over prolonged periods. To evaluate this, a field study was designed to examine the effects of very light-intensity lamb grazing on grassland soil microbiomes with different soil backgrounds. No significant change (P > 0.05) was observed in any vegetation and soil variables. Nonetheless, hundreds of microbial functional gene families, but not bacterial taxonomy, were significantly (P < 0.05) shifted. The relative abundances of both taxonomic markers and functional genes related to nitrifying bacteria were also changed. Here, the observation highlighted herein, showing a high level of sensitivity with respect to functional traits (functionally categorized taxa or genes) in differentiating mild environmental disturbance, suggests that the key level at which to address microbial responses may not be "species" (by means of rRNA taxonomy), but rather at the functional gene level.

59 BASIC BIOLOGICAL SCIENCES↗