Dihydroagarofuran Derivatives from the Dried Roots of <i>Tripterygium wilfordii</i>

Five new sesquiterpene derivatives, including dihydroagarofuran pyridine macrolides <b>1</b>–<b>4</b> and dihydroagarofuran ester <b>18</b>, and 13 known dihydroagarofuran derivatives were isolated from the aqueous EtOH extract of the dried roots of <i>Tripterygium wilfordii</i>. An in vitro antiherpetic activity assay indicated that compounds <b>11</b> and <b>17</b> displayed weak and moderate inhibition against herpes simplex virus type II, respectively

Isolation and Identification of Antioxidant Compounds from <i>Gynura Bicolor</i> Stems and Leaves

<div><p>ABSTRACT</p><p>The antioxidant compounds in the stems and leaves of Gynura bicolor were studied. DPPH and ABTS radical scavenging assays were employed to evaluate antioxidant capacity. By solvent extraction and Sephadex LH-20 column chromatography in sequence, ethanol extracts of Gynura bicolor stems and leaves were fractionated to obtain their active fractions, which were further separated to obtain twelve compounds: <b>1-8</b> from stems, and <b>4, 8-12</b> from leaves. Their structures were elucidated on the basis of spectroscopic data (NMR and MS). Among these substances, compounds <b>1, 2, 3, 4</b>, and <b>8</b> with significant antioxidant activity were determined to be responsible active components for stems, and compounds <b>4, 8</b>, and <b>12</b> for leaves.</p></div

qRT-PCR validation of DEGs identified by RNA-Seq.

<p>Sixteen DEGs involved in the biosynthesis and metabolism of indican were randomly selected for qRT-PCR confirmation. The blue bars and red lines represent RNA-Seq RPKM data and qRT-PCR relative expression levels, respectively. The beta-actin gene was used as an internal control, and every experiment was repeated with three biological samples. All the values shown are the means ± SE.</p

Distribution of various types of SSRs identified in <i>B</i>. <i>cusia</i> unigenes.

<p>Distribution of various types of SSRs identified in <i>B</i>. <i>cusia</i> unigenes.</p

Histogram of KOG function classification of <i>B</i>. <i>cusia</i> unigenes.

<p>Histogram of KOG function classification of <i>B</i>. <i>cusia</i> unigenes.</p

<i>De novo</i> characterization of the <i>Baphicacanthus cusia</i>(Nees) Bremek transcriptome and analysis of candidate genes involved in indican biosynthesis and metabolism

<div><p><i>Baphicacanthus cusia</i> (Nees) Bremek is an herb widely used for the clinical treatment of colds, fever, and influenza in Traditional Chinese Medicine. The roots, stems and leaves can be used as natural medicine, in which indigo and indirubin are two main active ingredients. In this study, quantification of indigo, indirubin, indican and adenosine among various tissues of <i>B</i>. <i>cusia</i> was conducted using HPLC-DAD. Leaves have significantly higher contents than stems and roots (380.66, 315.15, 20,978.26, 4323.15 μg/g in leaves, 306.36, 71.71, 3,056.78, 139.45 μg/g in stems, and 9.31, 7.82, 170.45, 197.48 μg/g in roots, respectively). <i>De novo</i> transcriptome sequencing of <i>B</i>. <i>cusia</i> was performed for the first time. The sequencing yielded 137,216,248, 122,837,394 and 140,240,688 clean reads from leaves, stems and roots respectively, which were assembled into 51,381 unique sequences. A total of 33,317 unigenes could be annotated using the databases of Nr, Swiss-Prot, KEGG and KOG. These analyses provided a detailed view of the enzymes involved in indican backbone biosynthesis, such as cytochrome P450, UDP-glycosyltransferase, glucosidase and tryptophan synthase. Analysis results showed that tryptophan synthase was the candidate gene involved in the tissue-specific biosynthesis of indican. We also detected sixteen types of simple sequence repeats in RNA-Seq data for use in future molecular mark assisted breeding studies. The results will be helpful in further analysis of <i>B</i>. <i>cusia</i> functional genomics, especially in increasing biosynthesis of indican through biotechnological approaches and metabolic regulation.</p></div

Annotated CYP450, UGT, glucosidase, and tryptophan synthase DEG numbers among various tissues of <i>B</i>. <i>cusia</i>.

<p>The data in green bars show the down-regulated gene number and the data in red bars show the up-regulated gene number.</p

Venn diagram of unigene number annotated by BLASTx with the Nr, SwissProt, KOG and KEGG databases.

<p>Venn diagram of unigene number annotated by BLASTx with the Nr, SwissProt, KOG and KEGG databases.</p

Contents of four ingredients in leaves, stems and roots of <i>B</i>. <i>cusia</i>.

<p>The experiment was repeated three times. The amounts of indigo, indirubin, indican and adenosine in leaf tissue (380.66μg/g, 315.15 μg/g, 20,978.26 μg/g and 1224.12 μg/g, respectively) were higher than those in the stem tissue (306.36 μg/g, 71.71μg/g, 3056.78 μg/g and 139.45 μg/g, respectively, <i>p</i> = 0.000), the amounts of indigo, indirubin and indican in stem tissue were higher than those in the root tissue (9.31 μg/g, 7.82 μg/g and 170.45 μg/g, respectively, <i>p</i> = 0.000), and the amount of adenosine in stem tissue was lower than that in root tissue (197.48 μg/g), but the difference was not significant (<i>p</i> = 0.350).</p

Histogram of level 2 GO terms of the <i>B</i>. <i>cusia</i> unigenes.

<p>Histogram of level 2 GO terms of the <i>B</i>. <i>cusia</i> unigenes.</p