Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of flg22 treatment in Arabidopsis thaliana Ecotype Landsberg vs fls2-17 mutant

ABSTRACT: Effect of flg22 treatment in Arabidopsis thaliana Ecotype Landsberg versus fls2-17 mutant

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Edward Oakeley

PROVIDER: E-MEXP-81 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Arabidopsis systemic immunity uses conserved defense signaling pathways and is mediated by jasmonates.

Truman William W Bennett Mark H MH Kubigsteltig Ines I Turnbull Colin C Grant Murray M

Proceedings of the National Academy of Sciences of the United States of America 20070110 3

In the absence of adaptive immunity displayed by animals, plants respond locally to biotic challenge via inducible basal defense networks activated through recognition and response to conserved pathogen-associated molecular patterns. In addition, immunity can be induced in tissues remote from infection sites by systemic acquired resistance (SAR), initiated after gene-for-gene recognition between plant resistance proteins and microbial effectors. The nature of the mobile signal and remotely activ ...[more]

PMID: 17215350

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Project description:F1 hybrids can outperform their parents in yield and vegetative biomass, features of hybrid vigor which form the basis of the hybrid seed industry. The yield advantage of the F1 is lost in the F2 and subsequent generations. In Arabidopsis, from F2 plants which have a F1 âlike phenotype, we have by recurrent selection produced pure breeding F5/F6 lines âHybrid Mimicsâ, in which the characteristics of the F1 Hybrid are stabilized. These Hybrid Mimic lines, like the F1 Hybrid, have larger leaves than the parent plant, the leaves having increased photosynthetic cell numbers, and in some lines increased size of cells, suggesting an increased supply of photosynthate. A comparison of the differentially expressed genes in the F1 Hybrid with those of eight Hybrid Mimic lines has identified metabolic pathways altered in both; these pathways include down regulation of defense response pathways and altered abiotic response pathways. F6 Hybrid Mimic lines are mostly homozygous at each locus in the genome yet retain the large F1-like phenotype. Many alleles in the F6 plants, when they are homozygous, have expression levels different to the level in the parent. We consider this altered expression to be a consequence of trans-regulation of genes from one parent by genes from the other parent. Transregulation could also arise from epigenetic modifications in the F1. The pure breeding Hybrid Mimics have been valuable in probing the mechanisms of hybrid vigor and may also prove to be useful hybrid vigor equivalents in agriculture. Plant Materials: Seeds were sterilized and sown onto plates containing MSN medium (MS salts supplemented with 1% (w/v) sucrose, pH7 with KOH, 0.6% wt/vol Noble agar). After 2 days at 4Â°C, the plates were transferred to a growth room with conditions of 22 Â°C/18 Â°C (day/night) and 16h light/8h dark cycle under Philips Cool Daylight TLD 58W/840 fluorescent tubes providing a photosynthetic photon flux density of 130 - 150Î¼mol photons m-2Â·s-1 as measured using a quantum meter (Model MQ-200 calibrated for electric light source, Apogee). At 18 days after sowing (DAS), the plants were transferred to soil and grown to the reproductive stage in a growth room with controlled light conditions (OSRAM L 36W/865 LumLux Daylight, 130 - 150Î¼mol photons m-2Â·s-1). To minimize the edge effects, the positioning of both plates and trays on the shelves was rotated every two days. All the plants were grown under the condition described above unless specified. Plant sample preparation and RNA extraction: For the transcriptomes of 15 DAS plants, the aerial tissues of 15 day seedlings were sampled. For the transcriptomes of plants at 28 DAS, the four largest leaves of 28 day plants from the parental lines (C24, Ler), the reciprocal Hybrids and eight F4 Hybrid Mimic lines were harvested. Each sample comprised a pool of five plants. Two biological replicates of each sample were sequenced. Total RNA was isolated using QIAGEN RNeasy MiniKitTM following the product instructions. To eliminate variation in experimental conditions in different positions in the growth room, nine F4 plant lines were divided into two batches to ensure plants were grown on the same shelves in each experiment. Parental lines C24 and Ler and the F1 Hybrid from which the F4 lines derived from were grown under the same conditions in each experiment as controls. The first batch including samples C24 (RepA and RepB), Ler (RepA and RepB), C24 x Ler F1, F4-L1-1, F4-L1-2, F4-L2-1, F4-L2-2 and F4-S-1 were grown under the condition describe above, we also sampled the five smallest C24 seedlings and five smallest Ler seedlings among the population (n = 50); the mRNA sequencing was performed by the Australian Genome Research Facility (AGRF). The second batch including C24 (RepC and RepD), Ler (RepC and RepD), Ler x C24 F1 Hybrid, F4-L3-1, F4-L3-2, F4-L4-1 and F4-L4-2 were grown in the same light intensity as above but under different light tubes (SYLVANIA PREMIUM extra FL36W/865 Super Daylight deluxe light tube). For the transcriptome of the two F6 Hybrid Mimic lines at 15 DAS, C24, Ler, Ler x C24 F1 Hybrids and three siblings from each F6 line (L3-1-1-2 and L4-2-1-2) were grown on MS medium (MS salts supplemented with 1% (w/v) sucrose, pH 5.7 with KOH, add 0.6% wt/vol agar) with controlled light conditions (SYLVANIA PREMIUM extra FL36W/865 Super Daylight deluxe, 130 - 150Î¼mol photons m-2Â·s-1). The mRNA sequencing service was provided by AGRF on the Illumina platform, 100bp paired ends.

Project description:F1 hybrids can outperform their parents in yield and vegetative biomass, features of hybrid vigor which form the basis of the hybrid seed industry. The yield advantage of the F1 is lost in the F2 and subsequent generations. In Arabidopsis, from F2 plants which have a F1like phenotype, we have by recurrent selection produced pure breeding F5/F6 linesHybrid Mimics, in which the characteristics of the F1 Hybrid are stabilized. These Hybrid Mimic lines, like the F1 Hybrid, have larger leaves than the parent plant, the leaves having increased photosynthetic cell numbers, and in some lines increased size of cells, suggesting an increased supply of photosynthate. A comparison of the differentially expressed genes in the F1 Hybrid with those of eight Hybrid Mimic lines has identified metabolic pathways altered in both; these pathways include down regulation of defense response pathways and altered abiotic response pathways. F6 Hybrid Mimic lines are mostly homozygous at each locus in the genome yet retain the large F1-like phenotype. Many alleles in the F6 plants, when they are homozygous, have expression levels different to the level in the parent. We consider this altered expression to be a consequence of trans-regulation of genes from one parent by genes from the other parent. Transregulation could also arise from epigenetic modifications in the F1. The pure breeding Hybrid Mimics have been valuable in probing the mechanisms of hybrid vigor and may also prove to be useful hybrid vigor equivalents in agriculture. Plant Materials: Seeds were sterilized and sown onto plates containing MSN medium (MS salts supplemented with 1% (w/v) sucrose, pH7 with KOH, 0.6% wt/vol Noble agar). After 2 days at 4°C, the plates were transferred to a growth room with conditions of 22°C/18°C (day/night) and 16h light/8h dark cycle under Philips Cool Daylight TLD 58W/840 fluorescent tubes providing a photosynthetic photon flux density of 130 - 150 umol photons m-2·s-1 as measured using a quantum meter (Model MQ-200 calibrated for electric light source, Apogee). At 18 days after sowing (DAS), the plants were transferred to soil and grown to the reproductive stage in a growth room with controlled light conditions (OSRAM L 36W/865 LumLux Daylight, 130 - 150 umol photons m-2·s-1). To minimize the edge effects, the positioning of both plates and trays on the shelves was rotated every two days. All the plants were grown under the condition described above unless specified. Plant sample preparation and RNA extraction: For the transcriptomes of 15 DAS plants, the aerial tissues of 15 day seedlings were sampled. For the transcriptomes of plants at 28 DAS, the four largest leaves of 28 day plants from the parental lines (C24, Ler), the reciprocal Hybrids and eight F4 Hybrid Mimic lines were harvested. Each sample comprised a pool of five plants. Two biological replicates of each sample were sequenced. Total RNA was isolated using QIAGEN RNeasy MiniKitTM following the product instructions. To eliminate variation in experimental conditions in different positions in the growth room, nine F4 plant lines were divided into two batches to ensure plants were grown on the same shelves in each experiment. Parental lines C24 and Ler and the F1 Hybrid from which the F4 lines derived from were grown under the same conditions in each experiment as controls. The first batch including samples C24 (RepA and RepB), Ler (RepA and RepB), C24 x Ler F1, F4-L1-1, F4-L1-2, F4-L2-1, F4-L2-2 and F4-S-1 were grown under the condition describe above, we also sampled the five smallest C24 seedlings and five smallest Ler seedlings among the population (n = 50); the mRNA sequencing was performed by the Australian Genome Research Facility (AGRF). The second batch including C24 (RepC and RepD), Ler (RepC and RepD), Ler x C24 F1 Hybrid, F4-L3-1, F4-L3-2, F4-L4-1 and F4-L4-2 were grown in the same light intensity as above but under different light tubes (SYLVANIA PREMIUM extra FL36W/865 Super Daylight deluxe light tube). For the transcriptome of the two F6 Hybrid Mimic lines at 15 DAS, C24, Ler, Ler x C24 F1 Hybrids and three siblings from each F6 line (L3-1-1-2 and L4-2-1-2) were grown on MS medium (MS salts supplemented with 1% (w/v) sucrose, pH 5.7 with KOH, add 0.6% wt/vol agar) with controlled light conditions (SYLVANIA PREMIUM extra FL36W/865 Super Daylight deluxe, 130 - 150 umol photons m-2Â·s-1). The mRNA sequencing service was provided by AGRF on the Illumina platform, 100bp paired ends.

Project description:Experiment 1 Gene profiling upon biotic stress treatment Biological question : What are the genes (incuding microRNA precursors) that are differentially regulated upon flg22 treatment? Experiment description : 14-day old seedlings (Col-0 and fls2 KO) treated with 10uM of flg22 (flagellin- derived peptide) over a one hour timecourse. Experiment 2 Gene profiling in microRNA mutants Biological question : What are the genes (including miRNA precursors) that are differentially regulated in a set of microRNA mutants? Experiment description : Ler, dcl1-9, hen1-1 and hasty were grown for 12-day on MS solid medium ,seedlings were then transferred in MS liquid medium and harvested 2 days after. Experiment 3 Gene profiling in siRNA mutants Biological question : What are the genes (including miRNA precursors) that are differentially regulated in a set of siRNA mutants? Experiment description : Col-0, dcl2-1, dcl3-1 and rdr2-1 were grown for 12-day on MS solid medium ,seedlings were then transferred in MS liquid medium and harvested 2 days after. Experiment 4 Gene profiling in Agrobacterium-induced tumors and auxin-induced calli. Biological question : What are the genes (including miRNA precursors) that are differentially regulated in an Agrobacterium-induced tumors compared to auxin-induced calli? Experiment description : Col-0 plants were infected with wt Agrobacterium tumefaciens (strain A208) by stabbing the emerging stems. 18 days later, growing tumors were harvested as were the non-infected part of the stabbed-stem.In parallel, Col-0 seeds were germinated on Callus Induction Media (CIM) and 18 days later, calli were harvested. Experiment 5 Gene profiling in tasiRNA mutants Biological question : What are the genes (including miRNA precursors) that are differentially regulated in a set of tasiRNA mutants? Experiment description : Col-0, dcl4-1, rdr6-1 were grown for 12-day on MS solid medium ,seedlings were then transferred in MS liquid medium and harvested 2 days after. Experiment 6 Gene profiling upon biotic stress treatment Biological question : What are the genes (incuding microRNA precursors) that are differentially regulated upon flg22 treatment? Experiment description : 14-day old seedlings (Col-0 and fls2 KO) treated with 10uM of flg22 (flagellin- derived peptide) over a one hour timecourse.

Project description:Plant growth and development is continuous and modular, a combination that allows morphogenesis by controlled cell division and elongation and serves to facilitate plantsM-bM-^@M-^Y adaptations to changing environments. Plasticity is achieved through integration of external and internal signals to regulate meristem activities. We previously characterized the pleiotropic phenotypes of the harlequin (hlq) mutant, isolated on the basis of ectopic expression of ABA- and auxin-inducible proDc3:GUS in roots and hypocotyls. Mutants are skotomorphogenic (dark development of true leaves and chlorophyll accumulation), have deformed and collapsed epidermal cells that accumulate callose and starch and stain strongly with the vital dye fluorescein diacetate, brittle stems and leaves, cell walls with a relative abundance of pectins and cell wall proteins, and abnormal and reduced root hairs and leaf trichomes. We cloned hlq and show it is an allele of brassinosteroid-insensitive3/hypocotyl6/root hairless3/Topoisomerase6B. Mutants affecting the cell wall and inhibitors of cellulose synthesis and N-glycosylation phenocopy several of the traits of hlq such as ectopic proDc3:GUS expression, lignin accumulation, and bulging root tips and epidermal cells. We performed a microarray analysis of genes differentially expressed in hlq mutants and observed coordinated expression of adjacent pairs/sets of genes suggesting a direct effect on chromatin remodeling, and significantly mis-regulated genes for histones, WRKY and IAA/AUX transcription factors, aquaporins, ubiquin E3-ligase mediated proteolysis and autophagy, biotic stress response markers, as well as numerous other cellular and metabolic processes affecting carbon partitioning into secondary metabolites lignin and anthocyanin. Our results shed light on the molecular mechanisms underlying the det/cop/fus-like pleiotropic phenotypes of hlq and support a broader role for TopoVI regulation of chromatin remodeling that mediates cell wall metabolism, growth, and development in response to environmental and hormonal signals. Seven samples; four mutant, three wild type in a dual color competitive hybridization to four microarray slides. One 'quasi-technical' replicate slide made up of pools of the three biological replicates plus a fourth mutant replicate.

Dataset Information

Transcription profiling of flg22 treatment in Arabidopsis thaliana Ecotype Landsberg vs fls2-17 mutant

Publications

Arabidopsis systemic immunity uses conserved defense signaling pathways and is mediated by jasmonates.

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