RNA‐Seq

RNA - Seq
  • 文章类型: Journal Article
    社区生态学的一个重要焦点,包括入侵生物学,是研究功能性状多样性模式,以理清环境和生物相互作用的影响。然而,一个值得注意的局限性是,研究通常依赖于一组小型且易于测量的功能特征,这可能不会立即反映对不断变化的非生物或生物条件的持续生态反应,包括在分子或生理水平上发生的那些。我们探索了使用表达基因的多样性-功能基因组多样性(FGD)-来了解最近和正在进行的高山入侵的生态动态的潜力。我们根据沿相邻入侵和原始河床发生的26种植物的转录组数据对FGD进行了量化。我们使用RNA-seq方法来总结表达转录本的总数及其对功能类别的注释,并将其与功能性状多样性(FTD)进行了对比,这些特征是传统上在植物生态学中考虑的。我们在被入侵的社区发现了更大的FGD和FTD,与物种丰富度的差异无关。然而,从FGD的角度来看,功能分散的幅度大于从FTD的角度来看。比较同类外来-本地物种对之间的FGD,我们没有发现许多显著差异的基因的比例,其注释匹配的功能类别。尽管如此,与原始物种相比,在入侵群落中具有更高的相对丰度的本地物种倾向于在入侵群落中以显着水平表达更大比例的基因,这表明烟气脱硫的变化可能与群落组成的变化有关。从群落到物种水平的多样性模式的比较提供了对驱动入侵动态的过程和机制的补充见解。FGD有可能阐明生态多样性的隐秘变化,我们预见了未来在分类水平和宏观生态系统中扩展的有希望的途径。
    An important focus of community ecology, including invasion biology, is to investigate functional trait diversity patterns to disentangle the effects of environmental and biotic interactions. However, a notable limitation is that studies usually rely on a small and easy-to-measure set of functional traits, which might not immediately reflect ongoing ecological responses to changing abiotic or biotic conditions, including those that occur at a molecular or physiological level. We explored the potential of using the diversity of expressed genes-functional genomic diversity (FGD)-to understand ecological dynamics of a recent and ongoing alpine invasion. We quantified FGD based on transcriptomic data measured for 26 plant species occurring along adjacent invaded and pristine streambeds. We used an RNA-seq approach to summarize the overall number of expressed transcripts and their annotations to functional categories, and contrasted this with functional trait diversity (FTD) measured from a suite of characters that have been traditionally considered in plant ecology. We found greater FGD and FTD in the invaded community, independent of differences in species richness. However, the magnitude of functional dispersion was greater from the perspective of FGD than from FTD. Comparing FGD between congeneric alien-native species pairs, we did not find many significant differences in the proportion of genes whose annotations matched functional categories. Still, native species with a greater relative abundance in the invaded community compared with the pristine tended to express a greater fraction of genes at significant levels in the invaded community, suggesting that changes in FGD may relate to shifts in community composition. Comparisons of diversity patterns from the community to the species level offer complementary insights into processes and mechanisms driving invasion dynamics. FGD has the potential to illuminate cryptic changes in ecological diversity, and we foresee promising avenues for future extensions across taxonomic levels and macro-ecosystems.
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  • 文章类型: Journal Article
    目的:TagSeq是一种经济有效的方法,用于需要大量样本的基因表达研究。迄今为止,植物中的TagSeq研究仅限于具有高质量参考基因组的植物。我们在非模型植物中测试了TagSeq参考转录组的适用性,作为圣丽塔实验范围国家生态观测网(NEON)核心站点自然基因表达变异研究的一部分。
    方法:TagSeq的组织样本来自四个物种的多个个体(Boutelouaaristidoides和Eragrostislehmanniana[禾本科],铜绿苔藓[Amaranthaceae],和佛罗里达帕金森病[Fabaceae])在三个日期的两个地点(总共56个样品)。每个物种一个样品用于通过标准RNA-seq产生参考转录组。通过参考基因座的回收来评估TagSeq性能,标记比对的特异性,和样本之间的差异。
    结果:大部分标签与每个参考对齐并唯一映射。数万个基因座的表达模式是可量化的,揭示了所有物种表达的一致空间差异。
    结论:使用从头参考转录组的TagSeq是本研究中定量基因表达的有效方法。标签是高度基因座特异性的,并为四种非模型植物物种产生生物学信息谱。
    OBJECTIVE: TagSeq is a cost-effective approach for gene expression studies requiring a large number of samples. To date, TagSeq studies in plants have been limited to those with a high-quality reference genome. We tested the suitability of reference transcriptomes for TagSeq in non-model plants, as part of a study of natural gene expression variation at the Santa Rita Experimental Range National Ecological Observatory Network (NEON) core site.
    METHODS: Tissue for TagSeq was sampled from multiple individuals of four species (Bouteloua aristidoides and Eragrostis lehmanniana [Poaceae], Tidestromia lanuginosa [Amaranthaceae], and Parkinsonia florida [Fabaceae]) at two locations on three dates (56 samples total). One sample per species was used to create a reference transcriptome via standard RNA-seq. TagSeq performance was assessed by recovery of reference loci, specificity of tag alignments, and variation among samples.
    RESULTS: A high fraction of tags aligned to each reference and mapped uniquely. Expression patterns were quantifiable for tens of thousands of loci, which revealed consistent spatial differentiation in expression for all species.
    CONCLUSIONS: TagSeq using de novo reference transcriptomes was an effective approach to quantifying gene expression in this study. Tags were highly locus specific and generated biologically informative profiles for four non-model plant species.
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  • 文章类型: Journal Article
    Background Current mammalian models for heart regeneration research are limited to neonatal apex amputation and myocardial infarction, both of which are controversial. RNAseq has demonstrated a very limited set of differentially expressed genes between sham and operated hearts in myocardial infarction models. Here, we investigated in rats whether pressure overload in the right ventricle, a common phenomenon in children with congenital heart disease, could be used as a better animal model for heart regeneration studies when considering cardiomyocyte proliferation as the most important index. Methods and Results In the rat model, pressure overload was induced by pulmonary artery banding on postnatal day 1 and confirmed by echocardiography and hemodynamic measurements at postnatal day 7. RNA sequencing analyses of purified right ventricular cardiomyocytes at postnatal day 7 from pulmonary artery banding and sham-operated rats revealed that there were 5469 differentially expressed genes between these 2 groups. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analysis showed that these genes mainly mediated mitosis and cell division. Cell proliferation assays indicated a continuous overproliferation of cardiomyocytes in the right ventricle after pulmonary artery banding, in particular for the first 3 postnatal days. We also validated the model using samples from overloaded right ventricles of human patients. There was an approximately 2-fold increase of Ki67/pHH3/aurora B-positive cardiomyocytes in human-overloaded right ventricles compared with nonoverloaded right ventricles. Other features of this animal model included cardiomyocyte hypotrophy with no fibrosis. Conclusions Pressure overload profoundly promotes cardiomyocyte proliferation in the neonatal stage in both rats and human beings. This activates a regeneration-specific gene program and may offer an alternative animal model for heart regeneration research.
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  • 文章类型: Journal Article
    An essential part of gene expression is the coordination of RNA synthesis and degradation, which occurs in the same cellular compartment in bacteria. Here, we report a genome-wide RNA degradation study in Escherichia coli using RNA-seq, and present evidence that the stereotypical exponential RNA decay curve obtained using initiation inhibitor, rifampicin, consists of two phases: residual RNA synthesis, a delay in the interruption of steady state that is dependent on distance relative to the mRNA\'s 5\' end, and the exponential decay. This gives a more accurate RNA lifetime and RNA polymerase elongation rate simultaneously genome-wide. Transcripts typically have a single RNA decay constant along all positions, which is distinct between different operons, indicating that RNA stability is unlikely determined by local sequences. These measurements allowed us to establish a model for RNA processing involving co-transcriptional degradation, providing quantitative description of the macromolecular coordination in gene expression in bacteria on a system-wide level.
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