Probing the Existence of an Alternative Gibberellin Receptor(s) in Rice Aleurone Cells
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ABSTRACT: Many years after the discovery of GID1 as the GA receptor, the belief in the existence of GA-binding protein (GBP) in aleurone cells still persists. Actually, some recent reviews and textbooks still contain information about the possibility of an alternative GA perception mechanism mediated by GBPs. Thus, to finally clarify this issue, we examined the expression of GA-regulated genes using rice embryoless half-seeds of four kinds of GA signaling mutants, each one independently carrying mutated GID1, DELLA, and GA-related F-box, which are all encoded by single genes in rice, allowing for convenient study of the GA perception system. The comprehensive microarray analysis using these rice mutants revealed that all genes tested to be responsive to GA in the wild-type (WT) aleurone cells consistently did not respond to GA in the gid1 or slr1 mutant.
Project description:Many years after the discovery of GID1 as the GA receptor, the belief in the existence of GA-binding protein (GBP) in aleurone cells still persists. Actually, some recent reviews and textbooks still contain information about the possibility of an alternative GA perception mechanism mediated by GBPs. Thus, to finally clarify this issue, we examined the expression of GA-regulated genes using rice embryoless half-seeds of four kinds of GA signaling mutants, each one independently carrying mutated GID1, DELLA, and GA-related F-box, which are all encoded by single genes in rice, allowing for convenient study of the GA perception system. The comprehensive microarray analysis using these rice mutants revealed that all genes tested to be responsive to GA in the wild-type (WT) aleurone cells consistently did not respond to GA in the gid1 or slr1 mutant. All microarray experiments were performed according to the manufacturerM-bM-^@M-^Ys manual. The Feature Extraction software (Agilent Technologies) was used to delineate and measure the Cy3 and Cy5 signal intensities of each spot in the array. The resulting data were normalized using the Variance-stabilizing normalization (VSN) algorithm.
Project description:The phytohormone gibberellic acid (GA) is well known to promote seed germination in plants. One of its functions is to stimulate the production of hydrolytic enzymes in the aleurone and their secretion to the adjacent endosperm. The storage in the endosperm is thus degraded by these hydrolases into small molecules, which are utilized as nutrients for embryo growth to establish the young seedling. ABA in contrast plays antagonistic role to GA to keep seed in dormancy. Cereal aleurone has been established as a model system to investigate giberrellin (GA) and abscisic acid (ABA) responses. Using Barley 1 GeneChip, we examined the mRNA accumulation of over 22 000 genes in barley aleurone treated with GA, ABA, GA plus ABA, and sln1 mutant.
Project description:The phytohormone gibberellic acid (GA) is well known to promote seed germination in plants. One of its functions is to stimulate the production of hydrolytic enzymes in the aleurone and their secretion to the adjacent endosperm. The storage in the endosperm is thus degraded by these hydrolases into small molecules, which are utilized as nutrients for embryo growth to establish the young seedling. ABA in contrast plays antagonistic role to GA to keep seed in dormancy. Cereal aleurone has been established as a model system to investigate giberrellin (GA) and abscisic acid (ABA) responses. Using Barley 1 GeneChip, we examined the mRNA accumulation of over 22 000 genes in barley aleurone treated with GA, ABA, GA plus ABA, and sln1 mutant. Barley aleurone tissues were separated from half-seed without embryo. Three independent RNA samples for each treatment including the control without any hormone were extracted and hybridized onto Affymetrix microarrays. We also did microarray in three replications for sln1 mutant aleurone without hormone treatment.
Project description:GA treatment of wild-type rice and use of DELLA protein mutant for RNA-seq and ChIP-seq. Explaining the chromatin mechanisms of DELLA-mediated gene repression and GA signalling-induced gene activation.
Project description:Microarray analysis was performed to know how many gibberellin (GA)-responsive genes are inhibited by beta-Yariv reagent, a specific binder of plant arabinogalactan-proteins. cRNAs were prepared from mRNAs isolated from aleurone protoplasts that were treated with GA, GA plus beta-Yariv reagent, or mock (DMSO)-treated for 24 hours, and were subjected to microarray analysis. The analysis was performed twice using target cRNAs prepared independently. Keywords: repeat sample
Project description:Transcriptional profiling of developing rice endosperm at seven days after flowering comparing aleurone layers with central starchy endosperm. Cereal productivity is dependent on the accumulation of storage compounds in the endosperm, a nutritive tissue that is composed of aleurone cells in the outermost regions and starchy endosperm in the inner region. The transcriptional analyses provides clues to the molecular basis for different metabolic pathways in response to the spatial and nutritional differences between rice aleurone cells and starchy endosperm.
Project description:Transcriptional profiling of developing rice endosperm at seven days after flowering comparing aleurone layers with central starchy endosperm. Cereal productivity is dependent on the accumulation of storage compounds in the endosperm, a nutritive tissue that is composed of aleurone cells in the outermost regions and starchy endosperm in the inner region. The transcriptional analyses provides clues to the molecular basis for different metabolic pathways in response to the spatial and nutritional differences between rice aleurone cells and starchy endosperm. Two-condition experiment, Aleurone layers vs. central starchy endosperm. 3 biological replicates with color swap for each biological replicate
Project description:Microarray analysis was performed to know how many gibberellin (GA)-responsive genes are inhibited by beta-Yariv reagent, a specific binder of plant arabinogalactan-proteins. cRNAs were prepared from mRNAs isolated from aleurone protoplasts that were treated with GA, GA plus beta-Yariv reagent, or mock (DMSO)-treated for 24 hours, and were subjected to microarray analysis. The analysis was performed twice using target cRNAs prepared independently. Experiment Overall Design: 2 control (DMSO), 2 gibberellin and 2 gibberellin plus beta-Yariv reagent-treated biological replicates were analyzed at 24 h.
Project description:In this study, we investigated novel rice genes that are expressed in aleurone cells by RNA-seq. RNA-seq was performed on four samples: a control sample, and samples treated with ABA, GA, and a mixture of the two hormones.