第73回千葉医学会総会記事

Microarray data of genes with lower expression levels (<0.74, p < 0.05) in 35S:ROXY19 plants as compared to control plants and their response to TIBA in the Col-0 background. The table contains the gene identity (AGI), description, the mean expression values of four independent biological replicates of the genotypes Col-0 and 35S:ROXY19#8, the ratios (FC, fold changes, log2) of the transcript levels in the transgenic line with respect to Col-0 and the corresponding p-values, the ratios between Col-0 treated with 0.1 % DMSO and Col-0 treated with 0.1 mM TIBA/0.1 % DMSO and the corresponding p-values. Since the microarray analysis of the TIBA-treated plants was performed with the Affimetrix ATH1 gene chip, the list contains 301 and not 321 genes. Genes that are not induced by TIBA are shown in light grey. FC, fold changes. For TIBA induction, plants were grown for six to seven weeks on steamed soil (Archut, Fruhstorfer Erde, T25, Str1fein) in growth chambers with light intensity at 37 to 45 μmol photons m−2 s−1 at 22 °C and 60 % humidity. Eight plants were sprayed with either 0.1 mM TIBA/0.1 % DMSO or with 0.1 % DMSO and leaves were harvested after eight h. The experiment was repeated three times. (XLSX 205 kb

Additional file 2: Figure S2. of Drought stress in maize causes differential acclimation responses of glutathione and sulfur metabolism in leaves and roots

Alignment of cytosolic glutathione reductase 1 (GR1; At3g24710) and the plastid and mitochondria localized GR2 (At2g54660) from Arabidopsis thaliana with the single identified homologous GR protein sequence from maize (GenBank accession no. AJ006055) with the CLUSTALW software. The alignment of these sequences showed sequence identity of maize GR of approximately 52 % with GR1 and 78 % with GR2 from Arabidopsis. (PDF 198 kb

Additional file 4: Table S1. of Drought stress in maize causes differential acclimation responses of glutathione and sulfur metabolism in leaves and roots

Accession number, genome annotations (http://www.maizegdb.org/) and primers used for quantification of transcript steady levels by qRT-PCR of maize genes addressed in this study. (PDF 202 kb

Additional file 2: Table S2. of Ectopically expressed glutaredoxin ROXY19 negatively regulates the detoxification pathway in Arabidopsis thaliana

Microarray data of genes with lower expression levels (FC < 0.74, p < 0.05) in 35S:ROXY19 plants as compared to control plants. The table contains the gene identity (AGI), description, the mean expression values of four independent biological replicates of the genotypes Col-0, 35S:GRXC2, 35S:ROXY19 SSMS , 35S:ROXY19#8 and 35S:ROXY19#12, and the ratios (FC, fold changes, log2) of the transcript levels in the transgenic lines with respect to Col-0 and the corresponding p-values. (XLSX 305 kb

<i>fou8</i> is affected in glucosinolate synthesis.

<p>Col-0, <i>fou8</i>, <i>apk1 apk2</i>, and <i>fou8 apk1 apk2</i> plants were grown for 5 weeks in controlled environment room. The total content of <b>A</b> glucosinolates and <b>B</b> desulfo-glucosinolates was measured in leaves. <b>C</b> Total RNA was isolated from leaves and the transcript levels of six genes involved in glucosinolate synthesis was determined by quantitative RT-PCR. The qRT-PCR reactions were performed in triplicate for each biological sample. The values in Col-0 were set to 1 for all genes. Results are presented as means ± SE from six pools of three individual plants grown in two independent experiments. Different letters mark values significantly different at P<0.05; asterisks mark values significantly different from Col-0 at P<0.05.</p

Mineral content of Col-0 and <i>fou8</i>.

<p>Col-0 and <i>fou8</i> plants were grown for 30 days in soil in controlled environment room. Whole rosettes were harvested and the mineral levels were determined by X-ray fluorescence spectrophotometry as % of dry weight. Results from one of two independent experiments are presented as means ± SD from three individual plants. Values substantially different between the two genotypes (P<0.05) are marked by asterisks.</p

Metabolites quantification and supplementation.

<p>(A) Schematic representation of sulphur assimilation in <i>A</i>. <i>thaliana</i>. Colour squares above the metabolites represent the log2 value of the <i>msa1-1</i>/WT Col-0 ratio of the concentration of each metabolite. APR: APS reductase; APS: adenosine 5’-phosphosulfate; ATPS: ATP sulfurylase; CBL: cystathionine β-lyase; CGS: cystathionine γ-synthase; Cyst: cystathionine; γ-ECS: γ-glutamylcysteine synthetase; γ-GluCys: γ-glutamylcysteine; GSHS: glutathione synthetase; Hcy: homocysteine; MS: methionine synthase; OAS: O-acetylserine; OAS-TL: OAS(thiol)lyase; SAT: serine acetyltransferase; SAMS, S-adenosylmethionine synthetase; SiR: sulphite reductase; SHM: serine hydroxymethyltransferase. (B-G) Measurement of sulphur-related metabolites. Plants were grown on agar solidified MGRL media under S sufficient (S1500) or S deficient (S0) conditions. Metabolites were extracted from shoots and roots and quantified by HPLC. Data are presented as means ± SD (<i>n</i> = 3). *, <i>P</i> ≤ 0.05; **, <i>P</i> ≤ 0.01, Student’s <i>t</i> test. (H-I) The concentrations of SAM and MTA in the shoots and roots of WT Col-0 and <i>msa1-1</i> grown under S sufficient condition. (J) Total S in the shoots of WT Col-0 and <i>msa1-1</i> grown under S sufficient condition without (CK) or with SAM added to the growth medium. Data in (B-J) are presented as means ± SD (<i>n</i> = 3 in (B-G), <i>n</i> = 5 in (H-I), and <i>n</i> = 6 in (J)). * and ** in (B-J) indicate values significantly different between WT Col-0 and <i>msa1-1</i> mutant at <i>P</i> ≤ 0.05 and <i>P</i> ≤ 0.01, respectively (Student’s <i>t</i> test). DW, dry weight. CK, control.</p

Cytosine methylation levels at CG, CHG, and CHH and total cytosine sites in WT and <i>msa1-1</i>.

<p>Cytosine methylation levels at CG, CHG, and CHH and total cytosine sites in WT and <i>msa1-1</i>.</p

Sulfate uptake and flux through sulfate assimilation in <i>fou8</i> and related mutants.

<p>WT Col-0 and mutants <i>fou8</i>, <i>fou2</i>, and <i>aos</i> were grown for 3 weeks on MS-phytagel vertical plates in controlled environment room. The seedlings were incubated for four hours with their roots submerged in nutrient solution adjusted to sulfate concentration of 0.2 mM and supplemented with 6.7 μCi [³⁵S]sulfate. Shoot and root material was harvested separately, and the flux was determined as incorporation of ³⁵S from [³⁵S] sulfate to thiols and proteins. <b>A</b> sulfate uptake, <b>B</b> Percentage of ³⁵S transported to leaves from the [³⁵S]sulfate taken up, <b>C</b> relative flux through the sulfate assimilation in the leaves calculated as % of incorporation in thiols and proteins from total [³⁵S]sulfate taken up. Results are presented as means ± SE from six independent pools of 8 seedlings grown in two independent experiments. Values marked with an asterisk show significant (P≤0.05) difference from Col-0.</p

Expression analysis of Arabidopsis lines.

<p>The various Arabidopsis lines were grown for 5 weeks in controlled environment room. Total RNA was isolated from leaves and the transcript levels of genes involved in sulfur metabolism, glucosinolate synthesis, and jasmonate synthesis were determined by quantitative RT-PCR. The qRT-PCR reactions were performed in triplicate for each of the six independent biological samples from plants grown in two independent experiments. Results are presented as a heat map of relative mRNA levels compared to Col-0. For comparison, sulfate levels are presented in the same way on the far right.</p