Up-regulation of <i>miRNA172E</i> under drought conditions.

<div><p>Each experiment was done triple with similar results.</p> <p>(<b>A</b>) Change in <i>pri-miRNA172</i> levels under drought conditions( Ler-0). </p> <p>(<b>B</b>) Change in mature <i>miRNA172</i> levels under drought conditions in wild-type plants. * <i>P</i><0.05. </p> <p>(<b>C</b>) RT-PCR analysis of <i>Pri-miRNA172A</i> and <i>Pri-miRNA172E</i> in the <i>gi</i> mutant under drought and control conditions. </p> <p>(<b>D</b>) Changes in mature <i>miRNA172A/B</i> and <i>miRNA172E</i> levels under drought conditions in the <i>gi</i> mutant.</p> <p>DR: Drought treatment began from the 14day age. For the mature miRNA assay, samples were collected at the 8^th day of DR treatment.</p></div

Differential gene expression in wild type (WT) and <i>gi</i> mutants under drought conditions as measured by digital gene expression.

<div><p>(<b>A</b>) Differential gene expression in WT( Ler-0) and <i>gi</i> mutants under drought conditions. </p> <p>(<b>B</b>) Venn diagram of up- and downregulated genes in WT and <i>gi</i> mutants with and without drought treatment. </p> <p>(<b>C</b>) Differential expression of <i>WRKY</i> genes in <i>gi</i> and WT under CK (standard) and DR(drought) conditions. Red: upregulated in <i>gi</i> compared with WT; green: down-regulated in <i>gi</i> compared with WT. </p> <p>DR treatment began from10 day age and maintained for 10 days. </p></div

Flowering times of <i>Arabidopsis</i> wild-type (WT) and mutants of different flowering pathways under drought stress.

<div><p>(<b>A</b>) Early flowering of WT (Col-0 and Ler-0) plants under drought stress and long-day conditions. </p> <p>(<b>B</b>) Flowering times of mutants of the photoperiod (<i>gi</i>, <i>co</i>), autonomous (flc-3), and phytohormone (gai) pathways under drought stress and long-day conditions. </p> <p>(<b>C</b>) Flowering times of WT (Col-0) plants under drought stress and short-day conditions. </p> <p>(<b>D</b>) Counted flowering times (days) of plants with different genotypes under CK and DR conditions. * flowering significantly earlier under DR condition than under CK condition.</p> <p>DR : Drought treatment began from 10days before flowering.</p></div

Sc₂C₂@<i>D</i>_3<i>h</i>(14246)‑C₇₄: A Missing Piece of the Clusterfullerene Puzzle

Clusterfullerenes with variable carbon cages have been extensively studied in recent years. However, despite all these efforts, C₇₄ cage-based clusterfullerene remains a missing piece of the puzzle. Herein, we show that single-crystal X-ray crystallographic analysis unambiguously assigns the previously reported dimetallofullerene Sc₂@C₇₆ to a novel carbide clusterfullerene, Sc₂C₂@<i>D</i>_3<i>h</i>(14246)-C₇₄, the first experimentally proven clusterfullerene with a C₇₄ cage. In addition, Sc₂C₂@<i>D</i>_3<i>h</i>(14246)-C₇₄ was charaterized by mass spectrometry, ultraviolet–visible–near-infrared absorption spectroscopy, ⁴⁵Sc nuclear magnetic resonance, and cyclic voltammetry. Comparative studies of the motion of the carbide cluster in Sc₂C₂@<i>D</i>_3<i>h</i>(14246)-C₇₄ and Sc₂C₂@C_2<i>n</i> (<i>n</i> = 40−44) revealed that a combination of factors, involving both the shape and size of the cage, is crucial in dictating the cluster motion. Moreover, structural studies of <i>D</i>_3<i>h</i>(14246)-C₇₄ revealed that it can be easily converted to <i>C</i>_<i>s</i>(10528)-C₇₂ and <i>T</i>_<i>d</i>(19151)-C₇₆ cages via C₂ desertion/insertion and Stone–Wales transformation. This suggests that <i>D</i>_3<i>h</i>(14246)-C₇₄ might play an important role in the growth pathway of clusterfullerenes

Transcriptional levels of <i>WRKY</i> genes in wild type (Ler-0) and <i>gi</i> mutants under standard (CK, white rectangles) and drought (DR, black rectangles) conditions.

<p>Results are averages of three biological replicates. *, significantly different (<i>P</i><0.05) expression levels between <i>gi</i> mutants and wild-type plants under CK or DR. DR treatment began from10 day age and maintained for 10 days. </p

Phylogenetic analysis of <i>Arabidopsis</i><i>WRKY</i> genes used in this study and <i>WRKY</i> genes from <i>Hordeum vulgare</i>.

<p>Data were analyzed by the neighbor joining method. Annotations indicate the regulation of <i>Arabidopsis </i><i>WRKY</i> genes by <i>GI</i>. The number above each branch-point referred to the bootstrap value (maximum is 100), which implied the reliability of existing clades in the tree. The system has performed 1000 replicates to construct the phylogram. The number in each clade represented the percentages of success for constructing the existing clade. 0.1 means 10% substitution rate between two sequences. </p

Yeast two-hybrid system analysis of WRKY and TOE1.

<p>Using TOE1 as bait identified WRKY44 as a potential protein interactor. Selective plates lacking adenine, histidine, tryptophan, and leucine (–Ade, –His, –Trp, –Leu) and control plates lacking only tryptophan (–Trp) are shown. Empty vectors (BD) and expressed proteins (TOE1) are indicated. Plates were photographed after 4 d. Potential interactors exhibited positive galactosidase activity (blue).</p

The <i>gi</i> mutant is sensitive to drought stress.

<div><p>(<b>A</b>) The phenotypes of wild-type plants ( Ler-0) and <i>gi</i> mutants under drought stress. (<b>B</b>) Transpiration rates of wild type, <i>gi</i> and <i>miRNA172A</i> (A1-10) <i>/D</i> (D6-3) <i>/E</i> (E1-2, E38-6) over-expressing plants. </p> <p>(<b>C</b>) Water loss in wild type, <i>gi</i> mutants, and plants over-expressing <i>miRNA172A</i> (A1-10) <i>/D</i> (D6-3) <i>/E</i> (E1-2, E38-6).</p> <p>DR treatment began from10 day age and maintained for 10 days.</p></div

Transcriptional level of <i>WRKY20</i>, <i>WRKY44</i>, and <i>WRKY51</i> in <i>co</i> and <i>miRNA172</i>–over-expressing plants (miRNA172-OX) under standard (CK, white rectangles) and drought (DR, black rectangles) conditions.

<p>Controls for the co mutant and miRNA172-OX was <i>Col-</i>0, the wild type in their respective ecotype backgrounds. Results are averages of three biological repeats. * Significantly different (<i>P</i><0.05) expression between miRNA172-OX and WT under both CK and DR conditions. E1-2 line was used as miRNA172-OX. DR treatment began from10 day age and maintained for 10 days. </p

Abundance of mRNAs of flowering-time and circadian-clock–regulated genes in <i>Arabidopsis</i> under long-day control (CK) and drought (DR) conditions.

<div><p>The expressions of <i>GI</i> (<b>A</b>), <i>FKF1</i> (<b>B</b>), <i>CO</i> (<b>C</b>), <i>FT</i> (<b>D</b>) were analyzed by real time-PCR in Ler-0 plants grown in LDs. For each gene, the first peak on the first day under CK conditions was standardized to a level of 1. Open and closed bars along the horizontal axis represent light and dark periods, respectively, measured in hours from dawn. Each experiment was done twice with similar results.</p> <p>===/ /=== represents the 5-d recovery period with watering. * indicated a significant difference (P<0.05).</p> <p>DR: Drought treatment began from the 10^th day age and maintained for 10 days.</p></div