14 research outputs found

    Comparative Transcriptomics Reveals Jasmonic Acid-Associated Metabolism Related to Cotton Fiber Initiation

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    <div><p>Analysis of mutants and gene expression patterns provides a powerful approach for investigating genes involved in key stages of plant fiber development. In this study, lintless-fuzzless XinWX and linted-fuzzless XinFLM with a single genetic locus difference for lint were used to identify differentially expressed genes. Scanning electron microscopy showed fiber initiation in XinFLM at 0 days post anthesis (DPA). Fiber transcriptional profiling of the lines at three initiation developmental stages (-1, 0, 1 DPA) was performed using an oligonucleotide microarray. Loop comparisons of the differentially expressed genes within and between the lines was carried out, and functional classification and enrichment analysis showed that gene expression patterns during fiber initiation were heavily associated with hormone metabolism, transcription factor regulation, lipid transport, and asparagine biosynthetic processes, as previously reported. Further, four members of the allene-oxide cyclase (AOC) family that function in jasmonate biosynthesis were parallel up-regulation in fiber initiation, especially at -1 DPA, compared to other tissues and organs in linted-fuzzed TM-1. Real time-quantitative PCR (RT-qPCR) analysis in different fiber mutant lines revealed that <i>AOCs</i> were up-regulated higher at -1 DPA in lintless-fuzzless than that in linted-fuzzless and linted-fuzzed materials, and transcription of the <i>AOCs</i> was increased under jasmonic acid (JA) treatment. Expression analysis of JA biosynthesis-associated genes between XinWX and XinFLM showed that they were up-regulated during fiber initiation in the fuzzless-lintless mutant. Taken together, jasmonic acid-associated metabolism was related to cotton fiber initiation. Parallel up-regulation of <i>AOCs</i> expression may be important for normal fiber initiation development, while overproduction of <i>AOCs</i> might disrupt normal fiber development.</p></div

    Differentially expressed genes involved in JA biosynthesis and signal transduction.

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    <p>Mixtures of ovules and fibers at -1, 0, and 1 DPA are shown in x-axis. Error bars indicate ± SD. Significant differences at the 0.05 and 0.01 probability levels are represented by * and **, respectively. The expression level was derived from triplicate experiments. Three biological replicates were used for calculation.</p

    <i>AOCs</i> expression in different tissues and organs in TM-1.

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    <p>R, S, L, Pe, A, -3d, -1d, 0d, 1d, 5d, 10d, 20d, and 25d indicate root, stem, leaf, petal, anther, -3, -1, 0, 1 and 3 DPA ovules and 5, 10, 20 and 25 DPA fibers, respectively. Significant differences compared with the expression in root tissue at the 0.05 and 0.01 probability levels are represented by * and **, respectively. Error bars indicate ± SD of triplicate experiments. Three biological replicates were used for calculation.</p

    Specific GO terms enriched between XinFLM and XinWX at three time points.

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    <p>NA, no result</p><p>FDR, false discovery rate.</p><p>Specific GO terms enriched between XinFLM and XinWX at three time points.</p

    Number of differentially expressed genes within and between XinWX and XinFLM.

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    <p>Microarray analysis was performed on three replicates of each sample at three stages (represented by boxes) for both accessions. Numbers on or beside the line designate the number of upregulated genes (>2-fold, FDR <0.05) relative to the adjacent developmental stage or between two accessions. The total number of genes in each comparison is given in brackets.</p

    Comparison of gene expression ratios detected by microarray and RT-qPCR analyses.

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    <p>(A) Relative expression of 12 genes during fiber initiation. Mixtures of ovules and fibers at -1, 0, 1 DPA are shown. Error bars indicate ± SD of triplicate experiments. Three biological replicates were used for calculation. Significant differences at the 0.05 and 0.01 probability levels are represented by * and **, respectively. (B) Comparison of gene expression ratios from microarray and RT-qPCR analyses. Data from 12 genes at -1, 0, and 1 DPA from XinWX and XinFLM are shown. Log<sub>2</sub> of the microarray expression ratios (x-axis) are plotted against log<sub>2</sub> of the RT-qPCR expression ratios (y-axis).</p

    Catalog of specific GO terms statistically over-represented during fiber initiation stages of XinWX and XinFLM.

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    <p>NA, no result</p><p>FDR, false discovery rate.</p><p>Catalog of specific GO terms statistically over-represented during fiber initiation stages of XinWX and XinFLM.</p

    Comparison of <i>AOCs</i> expression in XinWX and XinFLM.

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    <p>Ovules at -1, 0 and 1 DPA are shown. Significant differences at the 0.05 and 0.01 probability levels are represented by * and **, respectively. Error bars indicate ± SD. The expression level was derived from triplicate experiments. Three biological replicates were used for calculation.</p

    Exendin-4 attenuates integrin-related adhesion protein impairment induced by H<sub>2</sub>O<sub>2</sub>.

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    <p>(A and B) mRNA expression levels of integrin β1 and αV in ADSCs with or without Exendin-4 under H<sub>2</sub>O<sub>2</sub> injury by qRT-PCR analysis. (C-G) Representative western blotting of p-FAK, p-Src, paxillin, vinculin, talin and caspase-3 expression levels in ADSCs with or without Exendin-4 under H<sub>2</sub>O<sub>2</sub> injury. ADSCs were pretreated with 50 nM Exendin-4 for 24 h and then treated with or without 30 µM H<sub>2</sub>O<sub>2</sub> for 12 h. *<i>p</i><0.05.</p

    Confocal laser microscopic images of immunofluorescence analysis of differentiation of transplanted ADSCs in vivo (n = 12/group).

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    <p>(A and B) Representative images of differentiated cardiomyocytes-like cells using anti-cardiac troponin-I (green, cTnI) revealed significant augmentation of enhanced mRFP (red)/cTnI double positive cardiomyocyte-like cells (white arrow) in Ex-ADSCs group (B) compared with ADSCs group (A). (C) Quantitative analysis of the ratio of differentiated cardiomyocytes-like cells. (D and E) Representative images of differentiated vessel specific cells using anti-α-SMA (green) revealed significant enhancement of mRFP (red)/α-SMA double positive vessel-specific cells (white arrow) in Ex-ADSCs group (E) compared with ADSCs group(D). (F) Quantitative analysis of the ratio of differentiated vessel specific cells. *<i>p</i><0.05. Inset shows the corresponding boxed area magnified.</p
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