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

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

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

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

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

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

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

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

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

Data_Sheet_1_Cultivation of Drought-Tolerant and Insect-Resistant Rice Affects Soil Bacterial, but Not Fungal, Abundances and Community Structures.DOC

<p>The impacts of rice varieties with stacked drought tolerance and insect resistance on soil microbiomes are poorly understood. Hence, the objective of this study was to investigate the effects resulting from the cultivation of the drought-tolerant and insect-resistant rice cultivar, Hanhui3T, on soil physical–chemical properties, and bacterial and fungal community composition. Soil samples of Hanhui3T and conventional rice varieties (Hanhui3 and Zhonghua11) were collected in triplicate at the booting stage, and bacterial and fungal population sizes and community structures were assessed using qPCR and Illumina MiSeq sequencing, respectively. The Bt protein concentration of Hanhui3T was significantly higher than that of Hanhui3 and Zhonghua11, while the pH of Hanhui3T was significantly lower. Bacterial population sizes and community composition were significantly different between Hanhui3T and Hanhui3 (or Zhonghua11), while no similar effects were observed for fungal communities. These differences suggest that the effect of Hanhui3T cultivation on bacterial community composition is stronger than the effect on fungal communities. Moreover, bacterial abundance was positively correlated to soil pH, while bacterial community structure variations were mainly driven by soil pH and Bt protein concentration differences. In conclusion, the abundances and structure of bacterial communities were altered in rhizosphere with Hanhui3T cultivation that changed soil pH and Bt protein concentrations, while fungal communities displayed no such responsiveness.</p