11 research outputs found

    Legislative Documents

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    Also, variously referred to as: House bills; House documents; House legislative documents; legislative documents; General Court documents

    Additional file 1: Table S1. of microRNA-dependent gene regulatory networks in maize leaf senescence

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    Summary of small RNA classes in the samples. Table S2. Expression abundance of the known miRNAs. Table S3. Regions of the target genes identified by degradome sequencing. Table S4. Primers used in miRNA and target gene validation. (DOCX 61 kb

    Additional file 2: Figure S1. of microRNA-dependent gene regulatory networks in maize leaf senescence

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    T-plot of miRNA target genes. Five different categories of T-plots are shown. The degradome tag distributions along the target mRNA sequence are exhibited. The red line represents the sliced target transcripts. (TIF 4679 kb

    Disease symptoms induced by RSV infection in WT, <i>NS3</i> OX, and <i>mNS3</i> OX rice lines.

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    <p>(A) Images of whole plants and details showing stunted or folded-leaf phenotypes of the wild-type, <i>NS3</i> OX#1, <i>NS3</i> OX#7, <i>mNS3</i> OX#1 and <i>mNS3</i> OX#4 rice plants. Scale bars = 15 cm (upper panel) and 5 cm (lower panel). (B) Detection of the RSV <i>CP</i> gene by RT-PCR. (C) Time course of RSV symptom development in the WT and <i>NS3</i> OX or <i>mNS3</i> OX transgenic plants. Values represent the percentage of RSV-infected plants at various days post inoculation (dpi). Thirty plants were used for each treatment. **, <i>P</i> ā‰¤ 0.01. Average (Ā± SD) values from three biological replicates are shown.</p

    Proposed model.

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    <p><b>NS3 represses the PTGS pathway.</b> Viral-derived long double-stranded RNAs (dsRNA) are processed into 21-nt siRNAs, the 21-nt siRNAs are loaded into AGO protein complex for cleavage of viral RNA genome RNAs. However, NS3 binds viral long dsRNA or siRNAs and inhibits siRNA production or the loading of siRNAs into AGO complex. Therefore, the PTGS pathway is blocked by NS3. <b>NS3 enhances the miRNA biogenesis pathway.</b> In mock-infected rice plants, a given pri-miRNA has a low probability of being recognized by dimeric DRB1 and then being processed into a mature miRNA by the processor complex. Consequently, the mature miRNA is insufficient to repress its target gene, which plays a crucial role in antiviral activity or development. Under RSV infection, pri-miRNA-bound NS3 interacts with DRB1 in sites required for DRB1 self-interaction and acts as a scaffold to regulate the association between DRB1 and the pri-miRNA. This increases the chance that this pri-miRNA will be processed into a mature miRNA. In addition, this compromises the expression of the target gene and enhances RSV pathogenicity.</p

    RSV symptoms and miRNAs induced by NS3 in rice.

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    <p>(A) Images of whole plants exhibiting stunted phenotypes. Scale bar = 15 cm. (B) Detection of the RSV <i>CP</i> gene by RT-PCR. (C) Images of RSV-infected leaves exhibiting chlorotic mottling and rolled-leaf phenotypes. Scale bar = 5 cm. (D) Detection of miRNA (miR168, 395, 398, 399, and 528) accumulations in healthy (mock-infected) and RSV-infected rice plants by northern blotting. Red star, 21-nt miR528. (E) Northern and western blot assay detection of miRNA (miR168 and miR395) accumulations and protein expression in mock-infected, RSV-infected (RSV), <i>NS2</i> OX#1, <i>NS3</i> OX#1, <i>SP</i> OX#1, and <i>NSvc4</i> OX#1 rice plants. In (D and E), U6 served as a loading control, the expression levels in the WT-Mock plants are set to a value of 1.0 and the expression levels in the other plants are relative to this reference value.</p

    NS3 interacts with OsDRB1, a pri-miRNA processing factor.

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    <p>The BiFC assay was conducted in <i>N</i>. <i>benthamiana</i> epidermal cells, mCherry is a nuclear localization marker fused with red florescent protein (RFP). (A) Results of a BiFC assay showing the interaction between NS3 and OsDRB1a. Scale bar = 0.1 Ī¼m. (B) Results of a co-immunoprecipitation analysis showing the interaction between NS3 and OsDRB1a. (C) The accuracy of mutation sites of mOsDRB1a. (D) Results of a BiFC assay showing the accuracy of interaction sites between OsDRB1a. Scale bar = 0.1 Ī¼m (E) Formation of OsDRB1a dimers <i>in vivo</i>. Total rice protein extracts from the WT and <i>OsDRB1</i>-knockdown lines were treated with ā€˜ā€˜nativeā€ buffer and detected using anti-HYL1antibodies. (F) Results of a BiFC assay showing the accuracy of interaction sites between NS3 and DRB1a. Scale bar = 0.1 Ī¼m.</p

    Expression of NS3 promotes the accumulation of several miRNAs and reduces the expression of their targets.

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    <p>(A) Mutation site of the mutant NS3 (mNS3). (B) Measurement of miRNA (miR168, 395, 398, 399, and 528) accumulation in the WT, <i>NS3</i> OX#1, and <i>mNS3</i> OX#1 rice lines by small RNA sequencing. (C) Detection of miRNA (miR168, 395, 398, 399, and 528) accumulations in mock-infected, RSV-infected (RSV), <i>NS3</i> OX#1, <i>NS3</i> OX#7, <i>mNS3</i> OX#1, and <i>mNS3</i> OX#4 rice plants by northern blotting. U6 served as a loading control, the expression levels in the WT-Mock plants are set to a value of 1.0 and the expression levels of in the other plants are relative to this reference value. (D) Relative expression levels of the target genes of the miRNAs (miR168, 395, 398, 399, and 528), including <i>OsAGO1a</i>, <i>OsSULTR2;1</i>, <i>OsCSD1</i>, <i>Os08g45000</i>, and <i>OsRFPH2-10</i> in the mock-infected, RSV-infected (RSV), <i>NS3</i> OX#1, <i>NS3</i> OX#7, <i>mNS3</i> OX#1, and <i>mNS3</i> OX#4 rice plants. (E) Relative expression levels of the miRNA (miR168, 395, 398, 399, and 528) precursors, including <i>pri-miR168a</i>, <i>pri-miR395d</i>, <i>pri-miR398a</i>, <i>pri-miR399b</i>, and <i>pri-miR528</i> in the mock-infected, RSV-infected (RSV), <i>NS3</i> OX#1, <i>NS3</i> OX#7, <i>mNS3</i>OX#1, and <i>mNS3</i> OX#4 rice plants. Average (Ā± SD) values based on RT-qPCR analysis of three biological replicates are shown. ***, <i>P</i> ā‰¤ 0.001; **, <i>P</i> ā‰¤ 0.01; *, <i>P</i> ā‰¤ 0.05.</p
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