Additional file 2: Figure S1. of Transcriptome-wide high-throughput deep m6A-seq reveals unique differential m6A methylation patterns between three organs in Arabidopsis thaliana

The m6A peak and adenosine peak deduced from the HPLC-MS/MS analysis. a The relative m6A peak height (upper) and adenosine peak height (lower) in the standard sample. b The relative m6A peak height (upper) and adenosine peak height (lower) in the input sample. c The relative m6A peak height (upper) and adenosine peak height (lower) in the RIP sample. (DOC 50 kb

Additional file 9: Table S8. of Transcriptome-wide high-throughput deep m6A-seq reveals unique differential m6A methylation patterns between three organs in Arabidopsis thaliana

The primers used for qRT-PCR. (DOC 35 kb

Additional file 4: Table S3. of Transcriptome-wide high-throughput deep m6A-seq reveals unique differential m6A methylation patterns between three organs in Arabidopsis thaliana

Number of m6A sites detected in the three organs of Arabidopsis. (DOC 37 kb

Additional file 11: Figure S3. of Transcriptome-wide high-throughput deep m6A-seq reveals unique differential m6A methylation patterns between three organs in Arabidopsis thaliana

RNA QC results of the total RNA and the RIP RNA for m6A-seq samples. a RNA quality for the total RNA sample was high with RIN over 8.5. b RNA fragmentation for the m6A-seq samples was consistent in the experiments, with an average length of 106 nt. (DOC 313 kb

The miR165/166 Mediated Regulatory Module Plays Critical Roles in ABA Homeostasis and Response in <i>Arabidopsis thaliana</i>

<div><p>The function of miR165/166 in plant growth and development has been extensively studied, however, its roles in abiotic stress responses remain largely unknown. Here, we report that reduction in the expression of miR165/166 conferred a drought and cold resistance phenotype and hypersensitivity to ABA during seed germination and post-germination seedling development. We further show that the ABA hypersensitive phenotype is associated with a changed transcript abundance of ABA-responsive genes and a higher expression level of <i>ABI4</i>, which can be directly regulated by a miR165/166 target. Additionally, we found that reduction in miR165/166 expression leads to elevated ABA levels, which occurs at least partially through the increased expression of <i>BG1</i>, a gene that is directly regulated by a miR165/166 target. Taken together, our results uncover a novel role for miR165/166 in the regulation of ABA and abiotic stress responses and control of ABA homeostasis.</p></div

PHB promotes <i>ABI4</i> expression by directly binding to its promoter.

<p>(A) <i>ABI4</i> expression was analyzed in wild type and STTM165/166 2-day-old seedlings using qRT-PCR. (B) <i>ABI4</i> expression was analyzed in <i>PHB</i>:<i>PHB G202G-YFP</i> lines using qRT-PCR. (C) Analysis of <i>ABI4</i> promoter. A 3.0kb fragment upstream of ATG was chosen for the promoter analysis. (D) EMSA assay showed that PHB binds to an <i>ABI4</i> promoter region. Labeled probe of the <i>ABI4</i> promoter region was incubated with GST-PHB fusion protein. For the competition test, a non-labeled probe was added at 10-fold and 100-fold concentrations.</p

PHB directly upregulates the expression of <i>BG1</i>.

<p>(A) <i>BG1</i> expression was analyzed using qRT-PCR in <i>PHB</i>:<i>PHB G202G-YFP</i> lines. (B) Analysis of <i>BG1</i> promoter. The diagram shows a 4.0kb fragment upstream of the first ATG codon. (C) ChIP-qPCR was performed using specific primers corresponding to different promoter regions. (D) EMSA assay showed that PHB binds to the <i>BG1</i> promoter region. Labeled probes of the <i>BG1</i> promoter region were incubated with GST-PHB fusion protein. For the competition test, a non-labeled probe was added at 10-fold and 100-fold concentrations.</p

STTM165/166 plants exhibit drought stress resistance phenotype.

<p>(A) Drought resistance phenotype. 3-week-old plants (upper panel) were grown under the same conditions but without irrigation for 14 days (middle panel), and then rewatered for 2 days (lower panel). (B) Quantification of the survival rate. Forty plants of wild type and STTM165/166 were used in each experiment, and the survival rate was calculated from the results of four independent experiments. (C) Water loss assay. Aerial parts of 3-week-old plants were detached and weighed at the indicated time points. Water content at any time point was calculated as percentage of the fresh weight at time zero. Data were derived from four independent experiments (±SD).</p

ABA content is altered in STTM165/166 plants.

<p>(A) Quantitative RT-PCR analysis of the expression of genes involved in ABA conjugation and de-conjugation. (B) Quantitative RT-PCR analysis of <i>BG1</i> expression in various tissues of wild type and STTM165/166 plants. (C) Comparison of the ABA content between wild type and STTM165/166 plants.</p

Fine Tuning Water States in Hydrogels for High Voltage Aqueous Batteries

Hydrogels are widely used as quasi-solid-state electrolytes in aqueous batteries. However, they are not applicable in high-voltage batteries because the hydrogen evolution reaction cannot be effectively suppressed even when water is incorporated into the polymer network. Herein, by profoundly investigating the states of water molecules in hydrogels, we designed supramolecular hydrogel electrolytes featuring much more nonfreezable bound water and much less free water than that found in conventional hydrogels. Specifically, two strategies are developed to achieve this goal. One strategy is adopting monomers with a variety of hydrophilic groups to enhance the hydrophilicity of polymer chains. The other strategy is incorporating zwitterionic polymers or polymers with counterions as superhydrophilic units. In particular, the nonfreezable bound water content increased from 0.129 in the conventional hydrogel to >0.4 mg mg–1 in the fabricated hydrogels, while the free water content decreased from 1.232 to ∼0.15 mg mg–1. As a result, a wide electrochemical stability window of up to 3.25 V was obtained with the fabricated hydrogels with low concentrations of incorporated salts and enhanced hydrophilic groups or superhydrophilic groups. The ionic conductivities achieved with our developed hydrogel electrolytes were much higher than those in the conventional highly concentrated salt electrolytes, and their cost is also much lower. The designed supramolecular hydrogel electrolytes endowed an aqueous K-ion battery (AKIB) system with a high voltage plateau of 1.9 V and contributed to steady cycling of the AKIB for over 3000 cycles. The developed supramolecular hydrogel electrolytes are also applicable to other batteries, such as aqueous lithium-ion batteries, hybrid sodium-ion batteries, and multivalent-ion aqueous batteries, and can achieve high voltage output