Additional file 1 of Hydrogen sulphide alleviates iron deficiency by promoting iron availability and plant hormone levels in Glycine max seedlings
Creators: name: Chen, Juan
name: Zhang, Ni-Na
name: Pan, Qing
name: Lin, Xue-Yuan
name: Shangguan, Zhouping
name: Zhang, Jian-Hua
name: Wei, Ge-Hong
Publisher: name: figshare
Year: 2020
Subjects: subject: Genetics
subject: FOS: Biological sciences
subject: Plant Biology

description: Additional file 1. Materials and Methods. Figure S1. H2S but not other sulphur- or sodium-containing compounds derived from NaHS contributed to increased chlorophyll concentration in iron deficiency Glycine max seedlings. Glycine max seedlings were treated with 100 μM of different sulphur compounds including NaHS, Na2S, Na2SO4, Na2SO3, NaHSO4, NaHSO3, and NaAC for 15 d under iron deficiency condition (−Fe, 1 μM Fe). Figure S2. Effect of NaHS on the nodule number and nodule dry weight of iron-deficient Glycine max plants. The 3-w-old Glycine max seedlings were treated with the nutrition solution containing 1 μM Fe(III)-EDTA or 50 μM Fe(III)-EDTA with or without 100 μM NaHS for 15 days. Error bars represents the mean ± SE. Columns labelled with different letters indicate significant differences with P &lt; 0.05. Figure S3. Effect of NaHS on net photosynthesis (Pn) (A), stomatal conductance (Gs) (B), intrancellular CO2 concentration (Ci) (C), and water use efficiency (WUE) (D) of iron-deficient Glycine max plants. The 3-w-old Glycine max seedlings were treated with the nutrition solution containing 1 μM Fe(III)-EDTA or 50 μM Fe(III)-EDTA with or without 100 μM NaHS for 15 days. Error bars represents the mean ± SE. –Fe, 1 μM Fe; −Fe + NaHS, seedlings treated with 100 μM NaHS and 1 μM Fe; +Fe, 50 μM Fe; +Fe + NaHS, seedlings treated with 100 μM NaHS and 50 μM Fe. Figure S4. Effect of NaHS on photosystem II (PSII) (A), electronic transport ratio (ETR) (B), Fv/Fm (C), and Fv`/Fm` (D) of iron-deficient Glycine max plants. The 3-w-old Glycine max seedlings were treated with the nutrition solution containing 1 μM Fe(III)-EDTA or 50 μM Fe(III)-EDTA with or without 100 μM NaHS for 15 days. Error bars represents the mean ± SE. Columns labelled with different letters indicate significant differences with P &lt; 0.05. –Fe, 1 μM Fe; −Fe + NaHS, seedlings treated with 100 μM NaHS and 1 μM Fe; +Fe, 50 μM Fe; +Fe + NaHS, seedlings treated with 100 μM NaHS and 50 μM Fe. Figure S5. Effect of NaHS on the Cu (A-C), Mn (D-F), and Zn (G-I) concentrations of iron-deficient Glycine max plants. The 3-w-old Glycine max seedlings were treated with the nutrition solution containing 1 μM Fe(III)-EDTA or 50 μM Fe(III)-EDTA with or without 100 μM NaHS for 15 days. Error bars represents the mean ± SE. Columns labelled with different letters indicate significant differences with P &lt; 0.05. Figure S6. The correlation test of transcriptome sequencing data of Glycine max roots. There are three biological repeats per treatment. –Fe, 1 μM Fe; −Fe + NaHS, seedlings treated with 100 μM NaHS and 1 μM Fe; +Fe, 50 μM Fe; +Fe + NaHS, seedlings treated with 100 μM NaHS and 50 μM Fe. Figure S7. Volcano plot showing DEGs in Glycine max seedling treated with NaHS under iron deficiency condition. Biological significant (log2 fold change) is depicted on the x axis and statistical significant (log10) is depicted on the y axis. Statistical significance was corrected at P &lt; 0.05. (A) –Fe + NaHS vs –Fe; (B) + Fe + NaHS vs + Fe; (C) –Fe vs + Fe; (D) + Fe + NaHS vs –Fe + NaHS. –Fe, 1 μM Fe; −Fe + NaHS, seedlings treated with 100 μM NaHS and 1 μM Fe; +Fe, 50 μM Fe; +Fe + NaHS, seedlings treated with 100 μM NaHS and 50 μM Fe. Figure S8. Significantly enriched gene ontology (GO) terms (P &lt; 0.05) in the gene expression numbers in the root of Glycine max treated with NaHS under iron deficiency condition. (A) the up-regulated genes under –Fe + NaHS vs –Fe condition; (B) the down-regulated genes under –Fe + NaHS vs –Fe condition; (C) the up-regulated genes under +Fe + NaHS vs + Fe condition; (D) the down-regulated genes under +Fe + NaHS vs + Fe condition. GO terms belong to biological processes, molecular functions, and cellular components were shown in green, blue, and organ, respectively. Table S1. Summary of transcriptome sequencing data of Glycine max roots treated with NaHS under iron deficiency condition. –Fe, 1 μM Fe; −Fe + NaHS, seedlings treated with 100 μM NaHS and 1 μM Fe; +Fe, 50 μM Fe; +Fe + NaHS, seedlings treated with 100 μM NaHS and 50 μM Fe. Table S2. Statistics of genes in different expression-level interval of Glycine max roots treated with NaHS under iron deficiency condition. –Fe, 1 μM Fe; −Fe + NaHS, seedlings treated with 100 μM NaHS and 1 μM Fe; +Fe, 50 μM Fe; +Fe + NaHS, seedlings treated with 100 μM NaHS and 50 μM Fe. Table S3. Sequences of forward and reverse primers used in qRT-PCR for gene expression analysis in roots of Glycine max seedling under iron deficiency condition. Table S4. Procedures of dsDNA synthesis used in qRT-PCR for gene expression analysis in roots of Glycine max seedlings under iron deficiency condition. Table S5. Iron assimilation-related gene expression levels using transcriptome in Glycine max roots treated with NaHS under iron deficiency condition. –Fe, 1 μM Fe; −Fe + NaHS, seedlings treated with 100 μM NaHS and 1 μM Fe; +Fe, 50 μM Fe; +Fe + NaHS, seedlings treated with 100 μM NaHS and 50 μM Fe. Table S6. Sulfur assimilation-related gene expression levels using transcriptome in Glycine max roots treated with NaHS under iron deficiency condition. –Fe, 1 μM Fe; −Fe + NaHS, seedlings treated with 100 μM NaHS and 1 μM Fe; +Fe, 50 μM Fe; +Fe + NaHS, seedlings treated with 100 μM NaHS and 50 μM Fe. Table S7. Plant hormones-related gene expression levels using transcriptome in Glycine max roots treated with NaHS under iron deficiency condition. –Fe, 1 μM Fe; −Fe + NaHS, seedlings treated with 100 μM NaHS and 1 μM Fe; +Fe, 50 μM Fe; +Fe + NaHS, seedlings treated with 100 μM NaHS and 50 μM Fe. Table S8. Organic acid-related gene expression levels using transcriptome in Glycine max roots treated with NaHS under iron deficiency condition. –Fe, 1 μM Fe; −Fe + NaHS, seedlings treated with 100 μM NaHS and 1 μM Fe; +Fe, 50 μM Fe; +Fe + NaHS, seedlings treated with 100 μM NaHS and 50 μM Fe.