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      Unravelling miRNA regulation in yield of rice ( Oryza sativa) based on differential network model

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          Abstract

          Rice ( Oryza sativa L.) is one of the essential staple food crops and tillering, panicle branching and grain filling are three important traits determining the grain yield. Although miRNAs have been reported being regulating yield, no study has systematically investigated how miRNAs differentially function in high and low yield rice, in particular at a network level. This abundance of data from high-throughput sequencing provides an effective solution for systematic identification of regulatory miRNAs using developed algorithms in plants. We here present a novel algorithm, Gene Co-expression Network differential edge-like transformation (GRN-DET), which can identify key regulatory miRNAs in plant development. Based on the small RNA and RNA-seq data, miRNA-gene-TF co-regulation networks were constructed for yield of rice. Using GRN-DET, the key regulatory miRNAs for rice yield were characterized by the differential expression variances of miRNAs and co-variances of miRNA-mRNA, including osa-miR171 and osa-miR1432. Phytohormone cross-talks (auxin and brassinosteroid) were also revealed by these co-expression networks for the yield of rice.

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          Protocol: a highly sensitive RT-PCR method for detection and quantification of microRNAs

          MicroRNAs (miRNAs) are a class of small non-coding RNAs with a critical role in development and environmental responses. Efficient and reliable detection of miRNAs is an essential step towards understanding their roles in specific cells and tissues. However, gel-based assays currently used to detect miRNAs are very limited in terms of throughput, sensitivity and specificity. Here we provide protocols for detection and quantification of miRNAs by RT-PCR. We describe an end-point and real-time looped RT-PCR procedure and demonstrate detection of miRNAs from as little as 20 pg of plant tissue total RNA and from total RNA isolated from as little as 0.1 μl of phloem sap. In addition, we have developed an alternative real-time PCR assay that can further improve specificity when detecting low abundant miRNAs. Using this assay, we have demonstrated that miRNAs are differentially expressed in the phloem sap and the surrounding vascular tissue. This method enables fast, sensitive and specific miRNA expression profiling and is suitable for facilitation of high-throughput detection and quantification of miRNA expression.
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            Gene regulatory network inference: data integration in dynamic models-a review.

            Systems biology aims to develop mathematical models of biological systems by integrating experimental and theoretical techniques. During the last decade, many systems biological approaches that base on genome-wide data have been developed to unravel the complexity of gene regulation. This review deals with the reconstruction of gene regulatory networks (GRNs) from experimental data through computational methods. Standard GRN inference methods primarily use gene expression data derived from microarrays. However, the incorporation of additional information from heterogeneous data sources, e.g. genome sequence and protein-DNA interaction data, clearly supports the network inference process. This review focuses on promising modelling approaches that use such diverse types of molecular biological information. In particular, approaches are discussed that enable the modelling of the dynamics of gene regulatory systems. The review provides an overview of common modelling schemes and learning algorithms and outlines current challenges in GRN modelling.
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              New mini- zincin structures provide a minimal scaffold for members of this metallopeptidase superfamily

              Background The Acel_2062 protein from Acidothermus cellulolyticus is a protein of unknown function. Initial sequence analysis predicted that it was a metallopeptidase from the presence of a motif conserved amongst the Asp-zincins, which are peptidases that contain a single, catalytic zinc ion ligated by the histidines and aspartic acid within the motif (HEXXHXXGXXD). The Acel_2062 protein was chosen by the Joint Center for Structural Genomics for crystal structure determination to explore novel protein sequence space and structure-based function annotation. Results The crystal structure confirmed that the Acel_2062 protein consisted of a single, zincin-like metallopeptidase-like domain. The Met-turn, a structural feature thought to be important for a Met-zincin because it stabilizes the active site, is absent, and its stabilizing role may have been conferred to the C-terminal Tyr113. In our crystallographic model there are two molecules in the asymmetric unit and from size-exclusion chromatography, the protein dimerizes in solution. A water molecule is present in the putative zinc-binding site in one monomer, which is replaced by one of two observed conformations of His95 in the other. Conclusions The Acel_2062 protein is structurally related to the zincins. It contains the minimum structural features of a member of this protein superfamily, and can be described as a “mini- zincin”. There is a striking parallel with the structure of a mini-Glu-zincin, which represents the minimum structure of a Glu-zincin (a metallopeptidase in which the third zinc ligand is a glutamic acid). Rather than being an ancestral state, phylogenetic analysis suggests that the mini-zincins are derived from larger proteins.
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                Author and article information

                Contributors
                yiding@whu.edu.cn
                lnchen@sibs.ac.cn
                wwang@mail.kiz.ac.cn
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                31 May 2018
                31 May 2018
                2018
                : 8
                : 8498
                Affiliations
                [1 ]ISNI 0000 0004 1792 7072, GRID grid.419010.d, State Key Laboratory of Genetic Resources and Evolution, , Kunming Institute of Zoology, Chinese Academy of Sciences, ; Kunming, 650223 China
                [2 ]ISNI 0000 0001 2331 6153, GRID grid.49470.3e, State Key Laboratory of Hybrid rice, College of Life Sciences, , Wuhan University, ; Wuhan, 430072 China
                [3 ]ISNI 0000 0004 0467 2285, GRID grid.419092.7, Key Laboratory of Systems Biology, Innovation Center for Cell Signaling Network, Institute of Biochemistry and Cell Biology, , Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, ; Shanghai, 200031 China
                [4 ]Institute of Food Crop of Yunan Academy of Agricultural Sciences, Longtou Street, North Suburb, Kunming, 650205 China
                [5 ]ISNI 0000 0001 0307 1240, GRID grid.440588.5, Center for Ecological and Environmental Sciences, , Northwestern Polytechnical University, ; Xi’an, 710072 China
                Article
                26438
                10.1038/s41598-018-26438-w
                5981461
                29855560
                e8460505-4210-4c59-a5c7-f11d95488efb
                © The Author(s) 2018

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 22 September 2017
                : 8 May 2018
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