Project description:Drought stress response is a complex trait regulated at multiple levels. In the past few years, molecular and genomic studies have shown that several drought responsive genes (DRGs) with various functions are induced by drought stresses, and that various transcription factors (TFs) are involved in the regulation of stress-inducible genes. In addition to those DRGs mentioned above, microRNAs (miRNAs) are important regulators of gene expression at the posttranscriptional level by repressing mRNA expression. There is a complex interplay between transcriptional and post-transcriptional regulation of drought response that has not been extensively characterized in tobacco. In order to fully understand DRGs (including TFs) and different roles of miRNAs involved in the stress response, we sequenced and analysed three Digital Gene Expression (DGE) libraries in roots from drought treated tobacco plants, and four small RNA populations in roots, stems and leaves from control or drought treated tobacco plants. We identified 276 candidate DRGs in tobacco with sequence similarities to 64 known DRGs from model plants and crops and about 40% were TFs including WRKY, NAC, ERF and bZIP families. Furthermore, Out of these tobacco DRGs, 54differentially expressed DRGs included 21 TFs, which belonged to 24 TFs families such as NAC (6), MYB (4), ERF (10) and bZIP (1). Additionally, we confirmed expression of 39 known miRNA families (122 members) and five conserved miRNA families, which showed differential regulation under drought stress. Targets of miRNAs were further surveyed based on a recently published study, in which ten targets were DRGs. Finally, an integrated gene regulatory network has been proposed for the molecular mechanisms of the response of tobacco roots to drought stress using differentially expressed DRGs, the changed expression profiles of miRNAs and their target transcripts as a basis base on previous studies. In general, our data provide valuable information for future studies of the molecular mechanisms underlying tobacco roots in response to drought resistance in tobacco and other plants. Three tobacco (Nicotiana tabacum L.) roots tag-based DGE libraries treated at three time points (0, 6 and 48 h) with 20% PEG6000 were generated using Illumina 1G technology. The samples for four small RNA libraries was used based on the result of physiological index measurement as follows: equal quantities (10 ug) of total RNA isolated from tobacco roots treated with two time points (6 and 48 h) were mixed together to construct the drought -treated small RNA library (Root-treat), and total RNA prepared from the control roots sample was used to construct the control small RNA library (Root-ck). In addition, we also constructed another two libraries (stem and leaf) from leaves and stems of control plants, respectively. These libraries were constructed using the Small RNA Sample Prep Kit (Illumina, San Diego, CA) and then sent for Solexa sequencing.
Project description:Background: A comprehensive transcriptome survey, or "gene atlas", provides information essential for a complete understanding of the genomic biology of an organism. We present an atlas of RNA abundance for 92 adult, juvenile and fetal cattle tissues and 3 cattle cell lines. Results: The Bovine Gene Atlas was generated from 7.2 million unique digital gene expression tag sequences (300.2 million total raw tag sequences), from which 1.59 million unique tag sequences were identified that mapped to the bovine genome accounting for 85% of the total raw tag abundance. Filtering these tags yielded 87,764 unique tag sequences that unambiguously mapped to 16,517 annotated protein-coding loci in the genome accounting for 45% of the total raw tag abundance. Clustering of tissues based on tag abundance profiles generally confirmed ontology classification based on anatomy. There were 5,429 constitutively expressed loci and 3,445 constitutively expressed unique tag sequences mapping outside annotated gene boundaries that represent a resource for enhancing current gene models. Physical measures such as inferred transcript length or antisense tag abundance identified tissues with atypical transcriptional tag profiles. We report for the first time the tissue specific variation in the proportion of mitochondrial transcriptional tag abundance. The Bovine Gene Atlas can be examined at http://www.agbase.msstate.edu/bovineatlas . Conclusions: The Bovine Gene Atlas is the deepest and broadest transcriptome survey of any livestock genome to date. Commonalities and variation in sense and antisense transcript tag profiles identified in different tissues facilitate the examination of the relationship between gene expression, tissue, and gene function. An atlas of mRNA abundance for 92 adult, juvenile and fetal cattle tissues and 3 cattle cell lines.
Project description:We have quantified gene expression in five tissues (brain, heart, kidney, liver and testis) from humans, chimpanzees and rhesus macaques using the Illumina NlaIII Digital Gene Expression (DGE) protocol. This dataset extends a previous microarray study by Khaitovich et al. (Khaitovich et al. 2005) with the rhesus macaque outgroup and complements other previously generated tissue transcriptome profiles from primates (Enard et al. 2002; Khaitovich et al. 2006; Somel et al. 2009; Babbitt et al. 2010; Blekhman et al. 2010; Wetterbom et al. 2010). contributor: Max Planck Institute for Evolutionary Anthropology, Deutscher Platz 6, D-04103 Leipzig, Germany Samples were obtained from brain (pre-frontal cortex), heart, kidney, liver, and testis tissues of male humans, chimpanzees and rhesus macaques. Illumina NlaIII DGE libraries for all samples were generated in tissue batches, randomizing species in library preparation and sequencing. Human samples originate from different, probably unrelated, individuals for each tissue. For chimpanzees and rhesus macaques the libraries for all tissues come from the same set of individuals and among these are individuals related at the half- and full-sibling level. Due to limited access to samples, the analysis could not be limited to individuals of similar age. Human individuals vary between 5 and 88 years of age, chimpanzees between 6 years and 35 years of age and rhesus macaques between 3 and 9 years of age.
Project description:The goals of this study are to compare differential gene expressions for Penicillium oxalicum wild type strain (WT), laeA knockout strain (M-NM-^TlaeA), creA knockout strain (M-NM-^TcreA), and double genes knockout strain (M-NM-^TlaeAM-NM-^TcreA). The deletion of laeA downregulated genes involved in oxidation- reduction process, alkaloid metabolic process, and transmembrane transport. We find the expression levels of seven secondary metabolism gene clusters (totally 28 clusters) were silenced inM-NM-^TlaeA. The deletion of creA upregulated genes involved in hydrolase activity, acting on glycosyl bonds. Many genes involved in conidiation were drastically regulated inM-NM-^TlaeAM-NM-^TcreA. This study provides the information that combined laeA and creA function are required in conidiation and hydrolase activity of P. oxalicum. Examination of differential gene expressions by digital gene expression tag profiling in Penicillium oxalicum wild type strain and three mutant strains (laeA knockout strain, creA knockout strain and double genes knockout strain). qRTM-bM-^@M-^SPCR validation was performed using SYBR Green assays.
Project description:The goals of this study are to compare differential gene expressions for Penicillium oxalicum wild type strain (WT), and laeA knockout strain (ΔlaeA) in different development phase. The deletion of laeA downregulated genes involved in oxidation- reduction process, alkaloid metabolic process, and transmembrane transport. We find the expression levels of seven secondary metabolism gene clusters (totally 28 clusters) were silenced inΔlaeA. This study provides the information that laeA function are required in conidiation and hydrolase activity of P. oxalicum. Examination of differential gene expressions by digital gene expression tag profiling in Penicillium oxalicum wild type strain and laeA knockout mutant strains in 24h and 60h in modified Czapek culture medium with 2% glucose as carbon resource. qRT–PCR validation was performed using SYBR Green assays.
Project description:Transcriptomic analysis of fungus Penicillium decumbens and brlA deletion strains in liquid medium and solid medium respectivelly Examination of differential gene expressions by Penicillium decumbens strains 114-2 and brlA deletion stains in liquid medium and solid medium
Project description:Maize plants defective for the AGO104 protein show defects in chromatin condensation during meiosis, and subsequent failure to segregate chromosomes. AGO104 is a member of the ARGONAUTE family of proteins. AGO104 accumulates specifically in somatic cells surrounding the female meiocyte, suggesting a mobile signal rather than cell-autonomous control. AGO104 is necessary for non-CG methylation of centromeric and knob repeat DNA. Digital Gene Expression Tag Profiling experiments using high-throughput sequencing show that AGO104 influences the transcription of many targets in the ovaries, with a strong effect on centromeric repeats. AGO104 is related to Arabidopsis AGO9, but while AGO9 acts to repress germ cell fate in somatic tissues, AGO104 acts to repress somatic fate in germ cells. Thus, female germ cell development in maize is dependent upon conserved small RNA pathways acting non-cell autonomously in the ovule. Interfering with this repression leads to apomixis-like phenotypes in maize. Profiling of gene expression in mutant versus wilt type ovaries.
Project description:The present study profiled and analyzed gene expression of the maize ear at four key developmental stages. Based on genome-wide profile analysis, we detected differential mRNA of maize genes. Some of the differentially expressed genes (DEGs) were predicted to be potential candidates of maize ear development. Several well-known genes were found with reported mutants analyses, such as, compact plant2 (ct2), zea AGAMOUS homolog1 (zag1), bearded ear (bde), and silky1 (si1). MicroRNAs such as microRNA156 were predicted to target genes involved in maize ear development. Antisense transcripts were widespread throughout all the four stages, and are suspected to play important roles in maize ear development. Thus, identification and characterization of important genes and regulators at all the four developmental stages will contribute to an improved understanding of the molecular mechanisms responsible for maize ear development. Seeds of the maize inbred line 18-599 (Maize Research Institute, Sichuan Agricultural University, Chengdu, China) were grown in a growth chamber at 24°C/18°C (day/night) with 12 h illumination per day. Ears were collected as described previously [10] at four developmental stages: the growth point elongation, spikelet differentiation, floret primordium differentiation, and the floret organ differentiation phases. In brief, ears were manually collected at the four developmental stages. All the samples were harvested and immediately frozen in liquid nitrogen, and stored at -80°C until used for RNA isolation.
Project description:We report the application of Solexa/Illumina's digital gene expression (DGE) sequencing approaches to investigate inactivated Vibrio harveyi--induced transcriptome changes in Lateolabrax japonicas, a non model vertebrate species. Totally 3.44 and 3.22 million raw tags were measured. Then, gene annotation was performed by tags mapping analysis and the 169,950 non-redundant consensus sequences from RNA-seq based transcriptome analysis were used as reference transcript database. Tag mapping indicated that Vibrio harveyi--challenged adult Lateolabrax japonicas express over 70% of all genes represented in transcript databases. Meanwhile, totally 1224 consensus sequences exhibited significant difference after the bacterial challenge, in which 1183 transcripts can be well annotated, while approximately 41 transcripts have low sequence homology to the existing known sequences in public databases, suggesting that they might be putative novel immune-relevant genes in Lateolabrax japonicus closely related to the immunity for bacterial challenge. Our present study would greatly benefit to give deep insight into the immunogenetics in fish species, and clinical application in fish diseases. Examination of differentially expressed transcripts in baterial- and mock challenged fish.
Project description:RNA isolated from draining tracheobronchial lymph nodes (TBLN) from 5-week old pigs, either clinically infected with a feral isolate of Pseudorabies virus or uninfected were interrogated using Illumina Digital Gene Expression Tag Profiling. Over 100 million tag sequences were observed, representing 4,064,189 unique 21-base sequences collected from TBLN at time points 1, 3, 6 and 14 days post-infection (dpi) RNA isolated from draining tracheobronchial lymph nodes (TBLN) from 5-week old pigs (% per group pooled), either clinically infected with feral isolate FS268 of Pseudorabies virus or uninfected at 1, 3, 6, and 14 days post inoculation. Over 100 million tag sequences were observed, representing 4,064,189 unique 21-base sequences.