Project description:Identification of key genes and pathways regulated by phytohormone ABA during maize seed maturation, using the ABA synthesis-deficient mutant (vp5) and regular maize (Vp5) developing embryos.
Project description:To gain more information about the Kyn inhibition of TAA1/TARs, we conducted a RNA-seq analysis to compare the genome-wide gene expression profiles of WT seedlings grown on MS medium or MS plus Kyn, and wei8-/- tar2+/- progeny grown on MS. We report that Kyn treatment largely mimics the loss of TAA1/TAR2 functions. RNA seq of 3 samples: Col-0 on MS medium, Col-0 on MS+Kyn medium, wei8 tar2(+/-) on MS medium.
Project description:According to the physiological change of the ear leaf, the developing ear leaf after pollination can be divided into three classes: mature leaves [ML, 0–14 days after pollination (DAP)]; early senescent leaves (ESL, 15–24 DAP); and later senescent leaves (LSL, 25–30 DAP). We harvested ear leaves at 12 DAP, 20 DAP, and 28 DAP, representing ML, ESL, and LSL, respectively. To identify genes involved in the leaf senescence process, we sequenced three cDNA libraries, ML (12 DAP), ESL (20 DAP), and LSL (28 DAP) using an Illumina HiSeqTM 2000.
Project description:Comparative proteomics of ER-Golgi membranes from etiolated coleoptiles of maize (Zea mays) and young leaves of Arabidopsis (Arabidopsis thaliana) isolated by flotation centrifugation demonstrate differences in the complement of synthases and glycosyl transferases unique to the type of wall made. Additionally, maize Golgi membranes enriched by flotation were separated further by free-flow electrophoresis (FFE), yielding 3009 proteins, of which 229 were known to function in cell wall synthesis or metabolism. A subset of proteins identified after flotation centrifugation were identified in Golgi membranes but failed to enrich in them. Individual classes of proteins associated with cell wall synthesis were asymmetrically distributed across the fractions of Golgi membranes separated by FFE.
Project description:Characterisation of the differences in transcript abundance between maize leaves that have been exposed to blue, red, or no light: B73 maize seeds were planted and grown in the dark for 9 days. Etiolated second leaves were clamped 5 cm from the leaf tip by a Licor 6800 device equipped with a multi-phase flash fluorometer head, which administered 100 µmol m-2s-1 of either 100% red or 100% blue light. The Licor was configured with flow rate 500 µmol s⁻¹, 400 µmol mol⁻¹ CO2, leaf temperature 28°C and 60% humidity. Fluorescence and gas exchange were measured every 15 minutes for 3 hours. Leaf samples were collected between 12.30am-2pm each day.
Project description:Myocardial damage caused for example by cardiac ischemia leads to ventricular volume overload resulting in increased stretch of the remaining myocardium. In adult mammals, these changes trigger an adaptive cardiomyocyte hypertrophic response which, if the damage is extensive, will ultimately lead to pathological hypertrophy and heart failure. Conversely, in response to extensive myocardial damage, cardiomyocytes in the adult zebrafish heart and neonatal mice proliferate and completely regenerate the damaged myocardium. We therefore hypothesized that in adult zebrafish, changes in mechanical loading due to myocardial damage may act as a trigger to induce cardiac regeneration. Based, on this notion we sought to identify mechanosensors which could be involved in detecting changes in mechanical loading and triggering regeneration. Here we show using a combination of knockout animals, RNAseq and in vitro assays that the mechanosensitive ion channel Trpc6a is required by cardiomyocytes for successful cardiac regeneration in adult zebrafish. Furthermore, using a cyclic cell stretch assay, we have determined that Trpc6a induces the expression of components of the AP1 transcription complex in response to mechanical stretch. Our data highlights how changes in mechanical forces due to myocardial damage can be detected by mechanosensors which in turn can trigger cardiac regeneration.
Project description:In this study, a small RNA library from maize seed 24 hours after imbibition was sequenced by the Solexa technology. A total of 11,338,273 reads were obtained. 1,047,447 total reads representing 431 unique sRNAs matched to known maize miRNAs. Further analysis confirmed the authenticity of 115 known miRNAs belonging to 24 miRNA families and the discovery of 167 novel miRNAs in maize. Both the known and the novel miRNAs were confirmed by sequencing of a second small RNA library constructed the same way as the one used in the first sequencing. We also found 10 miRNAs that had not been reported in maize, but had been reported in other plant species. All novel sequences had not been earlier described in other plant species. In addition, seven miRNA* sequences were also obtained. Putative targets for 106 novel miRNAs were successfully predicted. Our results indicated that miRNA-mediated gene expression regulation is present in maize imbibed seed. This study led to the confirmation of the authenticity of 115 known miRNAs and the discovery of 167 novel miRNAs in maize. Identification of novel miRNAs resulted in significant enrichment of the repertoire of maize miRNAs and provided insights into miRNA regulation of genes expressed in imbibed seed. Discovery of novel miRNAs involved in imbibed maize seed by deep sequencing 2 independent small RNA libraries
Project description:Light is a pivotal signal for plants’ growth and survival. The promptly responses are required and achieving by sophisticated transcriptomic adjustments, the promptly translation enhancement, and effective protein surveillance. However, the global role of alternative splicing regulations in response to the light is unknown. Through global surveys of alternative splicing profile and frequency calculation for individual events normalized to transcriptomic changes, we identify nearly half of transcript present at this stage are alternatively spliced. Through the unbiased event comparison in D and L4h mRNA, we reveal a clear impact of splicing control on 2,961 genes during early photomorphogenesis. Intron retention is the most frequently event under controls of light as discovered in other developmental tissues and stages of Arabidopsis. Cryptic exon is the most abundant event preserved only in the dark-grown Arabidopsis. Genes underlying the splicing and transcriptional control mostly not overlapped indicate the distinct impacts of two regulatory mechanisms. Through confirmation with a homemade microarray for their responsiveness of light, we conclude 1,792 genes as biological validated candidates. Through functional study for genes underlying light-mediated splicing controls, we explore new light-signaling components which mainly responsive in isoform abundance but steady state mRNA. Among examination both positive (RPR39, KH-RBP, RRM and RRC1) and negative (PIF4, AMI1, DNAJ, UKL3, TF-IIs-like, and ubknown-Golgi protein) factors are controlled by splicing. Through predictions for example isoform structures, PIF4-IR4, AMI1-IR7-IR8, and DNAJ-IR1 both possess PTC and possibly produce truncated coding protein. PIF4-IR4 becomes best illustration for the dysfunction for the negative regulator by light. Above PTC+ isoforms together with one third of retained introns translate that evident for NMD-insensitive retained introns and supplements for protein levels. The event identities and cis-motif signatures associated with light regulations confirm a given uniqueness for the gene expression regulation during Arabidopsis photomorphogenesis. mRNA and polysome-associated mRNA of 4 days old Arabidopsis seedling grown under the dark or with 4 h light-treatment were isolated and the transcriptome and splicing events profiles were surveyed by mRNAseq. Independent 3 biological replicates of dark and L4h mRNA were applied for two-color-microarray using custom designed alternative-splicing array from Agilent Technologies.
Project description:1, Using mRNA-Seq to get expression profiling of rrp6l1-2 mutant and Col-0 wild-type (WT); 2,Using MethylC-Seq to provide single-base resolution of DNA methylation status in rrp6l1-2 mutant; 3, Using small RNA-Seq(sRNA-Seq) to get small RNA profiling of rrp6l1-2 and WT mRNA-Seq: 2 samples examined, WT and rrp6l1-2 mutant; MethylC-Seq: 1 sample examined, rrp6l1-2 mutant; small RNA-Seq: 2 samples examined, WT and rrp6l1-2 mutant