Project description:Most northern insect species experience a period of developmental arrest, diapause, which enables them to survive over the winter and postpone reproduction until favorable conditions. We studied the timing of reproductive diapause and its long-term effects on the cold tolerance of Drosophila montana, D. littoralis and D. ezoana females in seasonally varying environmental conditions. At the same time we traced expression levels of 219 genes in D. montana using a custom-made microarray. We show that the seasonal switch to reproductive diapause occurs over a short time period, and that overwintering in reproductive diapause has long-lasting effects on cold tolerance. Some genes, such as Hsc70, Jon25bi and period, were upregulated throughout the diapause, while others, including regucalcin, couch potato and Thor, were upregulated only at its specific phases. Some of the expression patterns induced during the sensitive stage, when the females either enter diapause or not, remained induced regardless of the later conditions. qPCR analyses confirmed the findings of the microarray analysis in D. montana and revealed similar gene expression changes in D. littoralis and D. ezoana. The present study helps to achieve a better understanding of the genetic regulation of diapause and of the plasticity of seasonal responses in general. Custom made DNA microarray for Drosophila montana and D. virilis. Current experiment includes 8 samples (7 to 250 days old diapausing or non-diapausing D. montana females) with two or three biological replicates
Project description:Background: Vernicia fordii (Tung oil tree) is a popular tree for biodiesel production in China. Unfortunately, the devastating Fusarium wilt disease caused great losses in production. Its sister species (Vernicia montana) was shown resistant to this pathogen. Vernicia fordii and Vernicia montana are main Vernicia species. V. fordii owns superior oil and agronomic traits, but susceptible to tung wilt disease, while V. nontana is resistant the wilt. However, the plants response mechanism remains largely unknown. Method: To define the divergence of gene expression modules between the resistant and susceptible Vernicia species responding to tung wilt pathogen, we generated comparative transcriptome atlases of two species during the process of infection with the pathogen F. oxysporum by RNA Sequencing in three biological replicates. Results: We describe the de novo assembly of the transcriptome of V. fordii and V. montana, comprising 258,430 and 245,240 non-redundant transcripts with N50 of 1,776 and 2,452 respectively. About 44,310 pair of putatively one-to-one orthologous genes between V. fordii and V. montana transcriptoms were identified. Overall, a high proportion of orthologous genes shared remarkably similar expression mode between Vernicia species. K means clustering indicated 2 cluster appear opposite expression mode. The highly connected gene expression analysis were conducted among genes with significantly differential expression mode, and the result indicated D6PK and LRR-RLK CLAVATA2 were top hub genes and hub genes glycosyltransferase (Gts), GLABRA2, PERK15 and EREBP-like factor were significantly associated with pathologic grades in resistant V. montana. Moreover, the result showed the resistant is a crucial signaling network, where MAPK signaling pathway, Plant-pathogen interaction, Circadian rhythm, Calcium signaling pathway and apoptosis fulfill distinct function. Additionally, dozens of unigenes were validated by quantitative real-time PCR (qRT-PCR). The study provided insight into the resistance gene expression modules. Conclusion: We first conducted a system analyses of the dynamics of gene expression both in susceptible and resistant Vernicia species with pathogen F. oxysporum infection. The results will serve as the important foundation to further deeply validation the resistance mechanism and breeding of the woody biodiesel plants V. fordii and V. montana.
Project description:Next-Generation-Sequencing (NGS) technologies have led to important improvement in the detection of new or unrecognized infective agents, related to infectious diseases. In this context, NGS high-throughput technology can be used to achieve a comprehensive and unbiased sequencing of the nucleic acids present in a clinical sample (i.e. tissues). Metagenomic shotgun sequencing has emerged as powerful high-throughput approaches to analyze and survey microbial composition in the field of infectious diseases. By directly sequencing millions of nucleic acid molecules in a sample and matching the sequences to those available in databases, pathogens of an infectious disease can be inferred. Despite the large amount of metagenomic shotgun data produced, there is a lack of a comprehensive and easy-use pipeline for data analysis that avoid annoying and complicated bioinformatics steps. Here we present HOME-BIO, a modular and exhaustive pipeline for analysis of biological entity estimation, specific designed for shotgun sequenced clinical samples. HOME-BIO analysis provides comprehensive taxonomy classification by querying different source database and carry out main steps in metagenomic investigation. HOME-BIO is a powerful tool in the hand of biologist without computational experience, which are focused on metagenomic analysis. Its easy-to-use intrinsic characteristic allows users to simply import raw sequenced reads file and obtain taxonomy profile of their samples.
Project description:Next-Generation-Sequencing (NGS) technologies have led to important improvement in the detection of new or unrecognized infective agents, related to infectious diseases. In this context, NGS high-throughput technology can be used to achieve a comprehensive and unbiased sequencing of the nucleic acids present in a clinical sample (i.e. tissues). Metagenomic shotgun sequencing has emerged as powerful high-throughput approaches to analyze and survey microbial composition in the field of infectious diseases. By directly sequencing millions of nucleic acid molecules in a sample and matching the sequences to those available in databases, pathogens of an infectious disease can be inferred. Despite the large amount of metagenomic shotgun data produced, there is a lack of a comprehensive and easy-use pipeline for data analysis that avoid annoying and complicated bioinformatics steps. Here we present HOME-BIO, a modular and exhaustive pipeline for analysis of biological entity estimation, specific designed for shotgun sequenced clinical samples. HOME-BIO analysis provides comprehensive taxonomy classification by querying different source database and carry out main steps in metagenomic investigation. HOME-BIO is a powerful tool in the hand of biologist without computational experience, which are focused on metagenomic analysis. Its easy-to-use intrinsic characteristic allows users to simply import raw sequenced reads file and obtain taxonomy profile of their samples.
Project description:Eukaryotic chromosomes feature large regions of compact, transcriptionally-repressed heterochromatin hallmarked by the presence of Heterochromatin Protein 1 (HP1) family members. HP1 proteins play multi-faceted roles in directly shaping the properties of heterochromatin, and in vivo , HP1 tethering to individual gene promoters leads to epigenetic modifications and gene silencing. However, emergent properties of HP1 at supranucleosomal scales have been difficult to study in cells due to a lack of appropriate tools. Here, we develop CRISPR-Engineered Chromatin Organization (EChO), a novel approach for combining live cell CRISPR-based imaging with inducible and reversible large-scale recruitment of heterochromatin components to native chromatin in human cells. Using CRISPR-EChO, we demonstrate that human HP1α binding across tens of kilobases of genomic DNA leads to formation of novel contacts with other heterochromatin regions and reversibly compacts chromatin from a diffuse to condensed state. The condensed chromatin state exhibits delayed disassembly kinetics and represses transcription across over 600 kilobases. Collectively, these findings support a polymer model of HP1α-mediated chromatin regulation and highlight the utility of CRISPR-EChO in studying and manipulating supranucleosomal chromatin organization in living cells.
Project description:Multiomics of faecal samples collected from individuals in families with multiple cases of type 1 diabetes mellitus (T1DM) over 3 or 4 months. Metagenomic and metatranscriptomic sequencing and metaproteomics were carried out, as well as whole human genome sequencing. Phenotypic data is available.