Project description:Pot grown plants of Arabidopsis thaliana, Cardamine hirsuta, Cardamine pratensis, Rorippa palustris and Rorippa sylvestris where completely submerged under ambient light conditions. After 24 and 48 hours the shoots were harvested for expression analysis. Differential expression analysis, taking into account unsubmerged control plants revealed that the Rorippa genus had a pronounced down regulation of the cell cycle whereas the Cardamine had an attenuated response to submergence.
Project description:To study the genes regulated by transcription factor SQUAMOSA PROMOTER BINDING PROTEIN-LIKE7 (SPL7) in Arabidopsis thaliana, we conducted chromatin immunoprecipitation-based sequencing (ChIP-seq) in 35S:FALG-SPL7 transgenic plant and spl7 mutant, and genome-wide sequencing-based transcript profiling of Arabidopsis thaliana wild-type plants and of spl7 mutant under MS medium and MS supplement with 5µM CuSO4. These experiments led to the identification of genes that are direct target of SPL7 and genes differentially expressed in an SPL7-dependent or Cu-dependent manner. This study provides a framework for the identification of SPL7 regulated genes towards characterization of SPL7 in copper homeostasis.
Project description:We used RNA-seq to profile gene expression changes during flg22 activated pattern-triggered immunity in multiple Brassicaceae including Capsella rubella, Cardamine hirsuta and Eutrema salsugineum as well as in multiple Arabidopsis thaliana accessions. This allows comparative transcriptomics within and across species to investigate the evolution of stress-responsive transcrption changes in these species.
Project description:To study the genes regulated by transcription factor SQUAMOSA PROMOTER BINDING PROTEIN-LIKE7 (SPL7) in Arabidopsis thaliana, we conducted chromatin immunoprecipitation-based sequencing (ChIP-seq) in 35S:FALG-SPL7 transgenic plant and spl7 mutant, and genome-wide sequencing-based transcript profiling of Arabidopsis thaliana wild-type plants and of spl7 mutant under MS medium and MS supplement with 5µM CuSO4. These experiments led to the identification of genes that are direct target of SPL7 and genes differentially expressed in an SPL7-dependent or Cu-dependent manner. This study provides a framework for the identification of SPL7 regulated genes towards characterization of SPL7 in copper homeostasis. ChIP-Seq: Chromatin isolation was performed with 7-day-old whole seedlings of spl7 mutant (negative control) and 35S:FALG-SPL7 transgene (sample) grown on standard MS medium under continuous white light according to Bowler et al. (2004). The resuspended chromatin pellet was sonicated at 4°C with a Diagenode Bioruptor set at high intensity for 10 min (30 s on, 30 s off intervals). The DNA was sheared to an average size of approximately 500 bp. Chromatin was immunoprecipitated with FLAG antibody (Sigma) according to the Affymetrix Chromatin Immunoprecipitation Assay Protocol Rev.3. Reverse cross-linked DNA was purified and used for sequencing libraries construction following the manufacture’s protocol (Illumina). RNA-Seq: DNase I-treated total RNA was prepared from 7-day-old whole seedlings of wild type (Col-0) and spl7 mutant grown on standard MS medium and MS supplement with 5µM CuSO4 under continuous white light (WL). First- and second-strand cDNA were generated using SuperScript II and random hexamer primers. Double-stranded cDNA was fragmented by nebulization and used for mRNA library construction following the manufacturer’s protocol (Illumina). Tophat was used to map the sequence reads to the Arabidopsis genome and only uniquely mapped reads were used in subsequent analyses. mRNA profiles of 7-day-old whole seedlings of wild type (Col-0) or spl7 mutant grown on standard MS medium and MS supplement with 5µM CuSO4 under continuous white light (WL) were generated by deep sequencing using illumine HiSeq2000.
Project description:Here we investigate the function of CUC1(CUP-SHAPED COTYLEDON1) in the diversification of leaf forms between simple-leaved Arabidopsis thaliana and compound-leaved Cardamine hirsuta. CUC transcription factors are conserved regulators in leaf margin dissection and leaflet formation. ChCUC1, ChCUC2 and ChCUC3 function redundantly and are required for the leaflet formation in C. hirsuta. Recently we discovered that ChCUC1 has species species-specific expression in young leaves of C.hirsuta. Moreover, interspecies gene transfer of ChCUC1 allele into A.thaliana is sufficient to increase leaf complexity. On this basis, we hypothesize that redeployment of ChCUC1 in leaves contributes to the formation of leaflets instead of serrations. However, the mechanism underlying ChCUC1 regulating cell division, cell polarity, cytoskeleton and thus leaf marginal patterning remains elusive. To this end, we make use of chromatin immunoprecipitation sequencing(ChIP-seq), transcriptomic, comparative genetics and advanced imaging approaches to identify the downstream regulating genes of ChCUC1.
Project description:An Ac/Ds transposon tagged mutant population was screened for changes in visible fruit phenotypes. One line showed orange, yellow sectors in the fruit and was named Orange ripening (Orr), its transposase free offspring showed Mendelian segregation yielding red, yellow and orange fruit bearing plants in a ratio of 1:2:1. Crossing the an orange fruit plant line to the wild-type yielded only plants bearing yellow fruit. A cross between the yellow fruit bearing progeny yielded 26 plants having red and 17 plants having yellow fruit, suggesting an over-dominant allele. Using inverse PCR analysis showed an insertion in the putative subunit M of the tomato Ndh complex. Subsequently, an Orr Ds transposon excision line was recovered which only showed red pigmented fruit. Here, we describe microarray profiling of tomato fruits from wild-type, heterozygous and homozygous Orr insertion plants and from fruits harvested from the Orr excision line. Keywords: mutant wild type