Comprehensive compendium of Arabidopsis RNA-seq data, A

2020 Spring.Includes bibliographical references.In the last fifteen years, the amount of publicly available genomic sequencing data has doubled every few months. Analyzing large collections of RNA-seq datasets can provide insights that are not available when analyzing data from single experiments. There are barriers towards such analyses: combining processed data is challenging because varying methods for processing data make it difficult to compare data across studies; combining data in raw form is challenging because of the resources needed to process the data. Multiple RNA-seq compendiums, which are curated sets of RNA-seq data that have been pre-processed in a uniform fashion, exist; however, there is no such resource in plants. We created a comprehensive compendium for Arabidopsis thaliana using a pipeline based on Snakemake. We downloaded over 80 Arabidopsis studies from the Sequence Read Archive. Through a strict set of criteria, we chose 35 studies containing a total of 700 biological replicates, with a focus on the response of different Arabidopsis tissues to a variety of stresses. In order to make the studies comparable, we hand-curated the metadata, pre-processed and analyzed each sample using our pipeline. We performed exploratory analysis on the samples in our compendium for quality control, and to identify biologically distinct subgroups, using PCA and t-SNE. We discuss the differences between these two methods and show that the data separates primarily by tissue type, and to a lesser extent, by the type of stress. We identified treatment conditions for each study and generated three lists: differentially expressed genes, differentially expressed introns, and genes that were differentially expressed under multiple conditions. We then visually analyzed these groups, looking for overarching patterns within the data, finding around a thousand genes that participate in stress response across tissues and stresses

Impact of Extrinsic Incubation Temperature on Natural Selection During Zika Virus Infection of Aedes Aegypti and Aedes Albopictus

Arthropod-borne viruses (arboviruses) require replication across a wide range of temperatures to perpetuate. While vertebrate hosts tend to maintain temperatures of approximately 37°C-40°C, arthropods are subject to ambient temperatures which can have a daily fluctuation of \u3e 10°C. Temperatures impact vector competence, extrinsic incubation period, and mosquito survival unimodally, with optimal conditions occurring at some intermediate temperature. In addition, the mean and range of daily temperature fluctuations influence arbovirus perpetuation and vector competence. The impact of temperature on arbovirus genetic diversity during systemic mosquito infection, however, is poorly understood. Therefore, we determined how constant extrinsic incubation temperatures of 25°C, 28°C, 32°C, and 35°C control Zika virus (ZIKV) vector competence and population dynamics within Aedes aegypti and Aedes albopictus mosquitoes. We also examined fluctuating temperatures which better mimic field conditions in the tropics. We found that vector competence varied in a unimodal manner for constant temperatures peaking between 28°C and 32°C for both Aedes species. Transmission peaked at 10 days post-infection for Aedes aegypti and 14 days for Aedes albopictus. Conversely, fluctuating temperature decreased vector competence. Using RNA-seq to characterize ZIKV population structure, we identified that temperature alters the selective environment in unexpected ways. During mosquito infection, constant temperatures more often elicited positive selection whereas fluctuating temperatures led to strong purifying selection in both Aedes species. These findings demonstrate that temperature has multiple impacts on ZIKV biology, including major effects on the selective environment within mosquitoes