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Analysis of gene expression from fbn5-1 T-DNA insertion mutant of Arabidopsis

ABSTRACT: We used microarrays to detail the global programme of gene expression underlying cellularisation and identified distinct classes of up-regulated genes in fbn5-1 T-DNA insertion Arabidopsis mutant. Fibrillins are lipid-associated proteins in plastids and are ubiquitous in plants. Fibrillins are known to accumulate and sequester carotenoids in chromoplasts during the development of flowers and fruits; however, little is known about the function of fibrillins in leaf tissues. Here we identified a novel protein fibrillin5 (FBN5) that is essential for plastoquinone-9 (PQ-9) biosynthesis in Arabidopsis thaliana. We showed that homozygous fbn5-1 plants were seedling-lethal, and XVE:FBN5-B transgenic plants expressing low levels of FBN5-B showed a lower growth rate and were smaller than wild-type. In chloroplasts, FBN5-B physically interacts with solanesyl-diphosphate synthase (SPS) 1 and 2, which synthesize the solanesyl moiety of PQ-9. Plants containing defective FBN5-B accumulated less PQ-9 and its cyclized product, plastochromanol-8, which is consistent with mutant phenotypes showing lower photosynthetic performance and higher levels of hydrogen peroxide. The role of FBN5-B in the FBN5-B/dimeric SPS complex is discussed with respect to biosynthesis of the solanesyl moiety.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE68275 | GEO | 2017/04/30

SECONDARY ACCESSION(S): PRJNA282370

REPOSITORIES: GEO

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Project description:In plants, fatty acids are de novo synthesized predominantly in plastids fromacetyl-CoA. Although fatty acid biosynthesis has been biochemically well-studied, little isknown about the regulatory mechanisms of the pathway. Here, we show that overexpressionof the Arabidopsis (Arabidopsis thaliana) LEAFY COTYLEDON1 (LEC1) gene causesglobally increased expression of fatty acid biosynthetic genes, which are involved in keyreactions of condensation, chain elongation and desaturation of fatty acid biosynthesis. Inthe plastidial fatty acid synthetic pathway, over 58% of known enzyme-coding genes areupregulated in LEC1-overexpressing transgenic plants, including those encoding threesubunits of acetyl-CoA carboxylase, a key enzyme controlling the fatty acid biosynthesisflux. Moreover, genes involved in glycolysis and lipid accumulation are also upregulated.Consistent with these results, levels of major fatty acid species and lipids were substantiallyincreased in the transgenic plants. Genetic analysis indicates that the LEC1 function ispartially dependent on ABSCISIC ACID INSENSITIVE3, FUSCA3 and WRINKLED1 in theregulation of fatty acid biosynthesis. Moreover, a similar phenotype was observed intransgenic Arabidopsis plants overexpressing two LEC1-like genes of Brassica napus.These results suggest that LEC1 and LEC1-like genes act as key regulators to coordinate theexpression of fatty acid biosynthetic genes, thereby representing a promising target forgenetic improvement of oil-production plants. Experiment Overall Design: The pER8-LEC1 transgenic seedlings were germinated and grown in the presence of 10 ÂµM estradiol for 4 days. The control sample was germinated and grown under the identical conditions without estradiol but containing 0.1% DMSO. Total RNA was prepared from fresh or frozen plant materials using the RNAeasy Plant Mini Kit (Qiagen China, Beijing). The first strand cDNA was synthesized, and then hybridized with the ATH1 oligonucleotide chips as described by the manufactureâs instructions (Affymetrix). The microarray hybridization data were collected and analyzed using related R (http://www.r-project.org/) packages provided by Bioconductor (http://www.biocoductor.org/). In brief, genes differentially expressed between wild type and mutant plants were selected by first removing âabsentâ genes which were never detected to be expressed in the experiments, then a two-side t-test was applied to remaining genes in order to test the expression difference between wide type and mutant plants. To avoid multiple testing problems, raw p-values were then adjusted into False Discovery Rate (FDR) using Benjamini and Hochberg approach. Finally, differentially expressed genes were defined as those FDR less than 0.2. Functional analysis of differentially expressed genes was carried out using the biological process category of Arabidopsis Gene Ontology. The hierarchical map of GO annotation was constructed according to the ontology tree provided by the Gene Ontology website (http://www.geneontology.com) as described previously (Zheng and Wang, 2008). Ontology categories that are significantly enriched among differentially expressed genes (hypergeometric test and FDR less than 0.1) were displayed as boxes in the map.

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Analysis of gene expression from fbn5-1 T-DNA insertion mutant of Arabidopsis

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