The permafrost carbon inventory on the Tibetan Plateau : a new evaluation using deep sediment cores

Acknowledgements We are grateful for Dr. Jens Strauss and the other two anonymous reviewers for their insightful comments on an earlier version of this MS, and appreciate members of the IBCAS Sampling Campaign Teams for their assistance in field investigation. This work was supported by the National Basic Research Program of China on Global Change (2014CB954001 and 2015CB954201), National Natural Science Foundation of China (31322011 and 41371213), and the Thousand Young Talents Program.Peer reviewedPostprin

Hotspots of soil organic carbon storage revealed by laboratory hyperspectral imaging

Subsoil organic carbon (OC) is generally lower in content and more heterogeneous than topsoil OC, rendering it difficult to detect significant differences in subsoil OC storage. We tested the application of laboratory hyperspectral imaging with a variety of machine learning approaches to predict OC distribution in undisturbed soil cores. Using a bias-corrected random forest we were able to reproduce the OC distribution in the soil cores with very good to excellent model goodness-of-fit, enabling us to map the spatial distribution of OC in the soil cores at very high resolution (~53 × 53 µm). Despite a large increase in variance and reduction in OC content with increasing depth, the high resolution of the images enabled statistically powerful analysis in spatial distribution of OC in the soil cores. In contrast to the relatively homogeneous distribution of OC in the plough horizon, the subsoil was characterized by distinct regions of OC enrichment and depletion, including biopores which contained ~2–10 times higher SOC contents than the soil matrix in close proximity. Laboratory hyperspectral imaging enables powerful, fine-scale investigations of the vertical distribution of soil OC as well as hotspots of OC storage in undisturbed samples, overcoming limitations of traditional soil sampling campaigns

APPLICATIONS OF MACHINE LEARNING IN MICROBIAL FORENSICS

Microbial ecosystems are complex, with hundreds of members interacting with each other and the environment. The intricate and hidden behaviors underlying these interactions make research questions challenging – but can be better understood through machine learning. However, most machine learning that is used in microbiome work is a black box form of investigation, where accurate predictions can be made, but the inner logic behind what is driving prediction is hidden behind nontransparent layers of complexity. Accordingly, the goal of this dissertation is to provide an interpretable and in-depth machine learning approach to investigate microbial biogeography and to use micro-organisms as novel tools to detect geospatial location and object provenance (previous known origin). These contributions follow with a framework that allows extraction of interpretable metrics and actionable insights from microbiome-based machine learning models. The first part of this work provides an overview of machine learning in the context of microbial ecology, human microbiome studies and environmental monitoring – outlining common practice and shortcomings. The second part of this work demonstrates a field study to demonstrate how machine learning can be used to characterize patterns in microbial biogeography globally – using microbes from ports located around the world. The third part of this work studies the persistence and stability of natural microbial communities from the environment that have colonized objects (vessels) and stay attached as they travel through the water. Finally, the last part of this dissertation provides a robust framework for investigating the microbiome. This framework provides a reasonable understanding of the data being used in microbiome-based machine learning and allows researchers to better apprehend and interpret results. Together, these extensive experiments assist an understanding of how to carry an in-silico design that characterizes candidate microbial biomarkers from real world settings to a rapid, field deployable diagnostic assay. The work presented here provides evidence for the use of microbial forensics as a toolkit to expand our basic understanding of microbial biogeography, microbial community stability and persistence in complex systems, and the ability of machine learning to be applied to downstream molecular detection platforms for rapid and accurate detection

Randomized lasso links microbial taxa with aquatic functional groups inferred from flow cytometry

High-nucleic-acid (HNA) and low-nucleic-acid (LNA) bacteria are two operational groups identified by flow cytometry (FCM) in aquatic systems. A number of reports have shown that HNA cell density correlates strongly with heterotrophic production, while LNA cell density does not. However, which taxa are specifically associated with these groups, and by extension, productivity has remained elusive. Here, we addressed this knowledge gap by using a machine learning-based variable selection approach that integrated FCM and 16S rRNA gene sequencing data collected from 14 freshwater lakes spanning a broad range in physicochemical conditions. There was a strong association between bacterial heterotrophic production and HNA absolute cell abundances (R-2 = 0.65), but not with the more abundant LNA cells. This solidifies findings, mainly from marine systems, that HNA and LNA bacteria could be considered separate functional groups, the former contributing a disproportionately large share of carbon cycling. Taxa selected by the models could predict HNA and LNA absolute cell abundances at all taxonomic levels. Selected operational taxonomic units (OTUs) ranged from low to high relative abundance and were mostly lake system specific (89.5% to 99.2%). A subset of selected OTUs was associated with both LNA and HNA groups (12.5% to 33.3%), suggesting either phenotypic plasticity or within-OTU genetic and physiological heterogeneity. These findings may lead to the identification of system-specific putative ecological indicators for heterotrophic productivity. Generally, our approach allows for the association of OTUs with specific functional groups in diverse ecosystems in order to improve our understanding of (microbial) biodiversity-ecosystem functioning relationships. IMPORTANCE A major goal in microbial ecology is to understand how microbial community structure influences ecosystem functioning. Various methods to directly associate bacterial taxa to functional groups in the environment are being developed. In this study, we applied machine learning methods to relate taxonomic data obtained from marker gene surveys to functional groups identified by flow cytometry. This allowed us to identify the taxa that are associated with heterotrophic productivity in freshwater lakes and indicated that the key contributors were highly system specific, regularly rare members of the community, and that some could possibly switch between being low and high contributors. Our approach provides a promising framework to identify taxa that contribute to ecosystem functioning and can be further developed to explore microbial contributions beyond heterotrophic production

Machine learning applications in microbial ecology, human microbiome studies, and environmental monitoring

Advances in nucleic acid sequencing technology have enabled expansion of our ability to profile microbial diversity. These large datasets of taxonomic and functional diversity are key to better understanding microbial ecology. Machine learning has proven to be a useful approach for analyzing microbial community data and making predictions about outcomes including human and environmental health. Machine learning applied to microbial community profiles has been used to predict disease states in human health, environmental quality and presence of contamination in the environment, and as trace evidence in forensics. Machine learning has appeal as a powerful tool that can provide deep insights into microbial communities and identify patterns in microbial community data. However, often machine learning models can be used as black boxes to predict a specific outcome, with little understanding of how the models arrived at predictions. Complex machine learning algorithms often may value higher accuracy and performance at the sacrifice of interpretability. In order to leverage machine learning into more translational research related to the microbiome and strengthen our ability to extract meaningful biological information, it is important for models to be interpretable. Here we review current trends in machine learning applications in microbial ecology as well as some of the important challenges and opportunities for more broad application of machine learning to understanding microbial communities

Machine learning for microbial ecology: predicting interactions and identifying their putative mechanisms

Microbial communities are key components of Earth’s ecosystems and they play important roles in human health and industrial processes. These communities and their functions can strongly depend on the diverse interactions between constituent species, posing the question of how such interactions can be predicted, measured and controlled. This challenge is particularly relevant for the many practical applications enabled by the rising field of synthetic microbial ecology, which includes the design of microbiome therapies for human diseases. Advances in sequencing technologies and genomic databases provide valuable datasets and tools for studying inter-microbial interactions, but the capacity to characterize the strength and mechanisms of interactions between species in large consortia is still an unsolved challenge. In this thesis, I show how machine learning methods can be used to help address these questions. The first portion of my thesis work was focused on predicting the outcome of pairwise interactions between microbial species. By integrating genomic information and observed experimental data, I used machine learning algorithms to explore the predictive relationship between single-species traits and inter-species interaction phenotypes. I found that organismal traits (e.g. annotated functions of genomic elements) are sufficient to predict the qualitative outcome of interactions between microbes. I also found that the relative fraction of possible experiments needed to build acceptable models drastically shrinks as the combinatorial space grows. In the second part of my thesis work, I developed an algorithmic method for identifying putative interaction mechanisms by scoring combinations of variables that random forest uses in order to predict interaction outcomes. I applied this method to a study of the human microbiome and identified a previously unreported combination of microbes that are strongly associated with Crohn’s disease. In the last part of my thesis, I utilized a regression approach to first identify and then quantify interactions between microbial species relevant to community function. The work I present in this dissertation provides a general framework for understanding the myriad interactions that occur in natural and synthetic microbial consortia

A non-linear Granger-causality framework to investigate climate-vegetation dynamics

Satellite Earth observation has led to the creation of global climate data records of many important environmental and climatic variables. These come in the form of multivariate time series with different spatial and temporal resolutions. Data of this kind provide new means to further unravel the influence of climate on vegetation dynamics. However, as advocated in this article, commonly used statistical methods are often too simplistic to represent complex climate-vegetation relationships due to linearity assumptions. Therefore, as an extension of linear Granger-causality analysis, we present a novel non-linear framework consisting of several components, such as data collection from various databases, time series decomposition techniques, feature construction methods, and predictive modelling by means of random forests. Experimental results on global data sets indicate that, with this framework, it is possible to detect non-linear patterns that are much less visible with traditional Granger-causality methods. In addition, we discuss extensive experimental results that highlight the importance of considering non-linear aspects of climate-vegetation dynamics

Assessment of hydrological and seasonal controls over the nitrate flushing from a forested watershed using a data mining technique

A data mining, regression tree algorithm M5 was used to review the role of mutual hydrological and seasonal settings which control the streamwater nitrate flushing during hydrological events within a forested watershed in the southwestern part of Slovenia, characterized by distinctive flushing, almost torrential hydrological regime. The basis for the research was an extensive dataset of continuous, high frequency measurements of seasonal meteorological conditions, watershed hydrological responses and streamwater nitrate concentrations. The dataset contained 16 recorded hydrographs occurring in different seasonal and hydrological conditions. Based on predefined regression tree pruning criteria, a comprehensible regression tree model was obtained in the sense of the domain knowledge, which was able to adequately describe most of the streamwater nitrate concentration variations (RMSE=1.02mg/l-N; r=0.91). The attributes which were found to be the most descriptive in the sense of streamwater nitrate concentrations were the antecedent precipitation index (API) and air temperatures in the preceding periods. The model was most successful in describing streamwater concentrations in the range 1-4 mg/l-N, covering large proportion of the dataset. The model performance was little worse in the periods of high streamwater nitrate concentration peaks during the summer hydrographs (up to 7 mg/l-N) but poor during the autumn hydrograph (up to 14 mg/l-N) related to highly variable hydrological conditions, which would require a less robust regression tree model based on the extended dataset