Additional file 1 of Strategies of chemolithoautotrophs adapting to high temperature and extremely acidic conditions in a shallow hydrothermal ecosystem
Creators: name: Deng, Wenchao
name: Zhao, Zihao
name: Li, Yufang
name: Cao, Rongguang
name: Chen, Mingming
name: Tang, Kai
name: Wang, Deli
name: Fan, Wei
name: Hu, Anyi
name: Chen, Guangcheng
name: Chen, Chen-Tung Arthur
name: Zhang, Yao
Publisher: name: figshare
Year: 2023
Subjects: subject: Biochemistry
subject: Physiology
subject: FOS: Biological sciences
subject: Environmental Sciences not elsewhere classified
subject: Chemical Sciences not elsewhere classified
subject: Ecology
subject: Biological Sciences not elsewhere classified
subject: Inorganic Chemistry
subject: FOS: Chemical sciences

description: Additional file 1: Supplementary figures. Fig. S1. Geographic location of Kueishantao Islet and geochemical characteristics of the white vent (WV) and yellow vent (YV). Fig. S2. Normalized distribution (data scaled between 0 and 1 along the gradient) of bacterial 16S rRNA gene copies in cesium chloride (CsCl) density gradients of temperature gradient incubated samples at the white vent (WV) and yellow vent (YV). The blue, pink, and red bars respectively represent the density range for the light (L, unlabeled), heavy (H, labeled with 13C), and ultra-heavy (UH, labeled with both 13C and 15N) DNA. The triangle, circle, and square symbols represent the fractions that were selected for high-throughput sequencing to obtain bacterial populations that incorporated both NaH13CO3 and 15NH4Cl, incorporated only NaH13CO3, and did not incorporate any labeled substrates, respectively. Fig. S3. Quantitative PCR tested bacterial (solid bar) and archaea (hollow bar) abundance in incubated samples and in in situ samples. WV, White vent; YV, yellow vent. Fig. S4. Nonmetric multidimensional scaling ordination based on Bray-Curtis dissimilarities among (a) bacterial 16S rRNA gene communities or (b) KEGG functional compositions of the ultra-heavy (UH), heavy (H) and light (L) fractions from temperature gradient incubated samples at White Vent (WV, circle) and Yellow Vent (YV, triangle). Each symbol represents an individual community. Fig. S5. Relative abundance of bacterial 16S rRNA gene reads assigned to phylogenetic orders. Fig. S6. Comparison of KEGG Orthologys (KOs) among 45 °C at site WV and 30 °C and 45 °C at site YV. In (a), circle size represents the highest normalized relative abundance (RB) of each KO among the three metagenomic libraries. If the difference in relative abundance of a KO was less than two-fold across the three metagenomic libraries, it was noted as a core function (gray circle). If the relative abundance of a KO in one metagenomic library was at least two-fold higher than its abundance in the remaining two libraries, the KO was noted as an enriched (Enr.) function (closed color circles) in that library. Functions with lower relative abundance in one metagenomic library compared to the other two libraries were classified as “diluted (Dil)” KOs (open circles) in that library. In (b), the total relative abundance of each KO category is shown, with the number of KOs in each category indicated above the bars. Fig. S7. Composition of core functions from the UH fraction in KEGG level 2 metabolic pathway. (a), comparison among WV 65 °C, WV 45 °C, and WV 30 °C; (b), comparison among YV 45 °C, YV 30 °C, and WV 30 °C. Fig. S8. Composition of enriched functions in the KEGG level 2 metabolic pathway from the ultra-heavy (UH) fraction. (a) Comparison among WV 65 °C, WV 45 °C, and WV 30 °C. (b) Comparison among YV 45 °C, YV 30 °C, and WV 30 °C. Fig. S9. Ternary plot comparing the abundance of genes encoding all subunits of proton-pumping NADH: ubiquinone oxidoreductase in the ultra-heavy (UH) fractions (a) among 65 °C, 45 °C and 30 °C at site WV, and (b) among WV 30 °C, YV 30 °C, and YV 45 °C. Circle size represents the highest normalized relative abundance (RB) of each gene among the three metagenomic libraries. If the difference in relative abundance of a gene was less than two-fold across the three metagenomic libraries, it was noted as a core gene (gray circle). If the relative abundance of a gene in one metagenomic library was at least two-fold higher than its abundance in the remaining two libraries, the gene was noted as an enriched (Enr.) gene (closed color circles) in that library. Genes with lower relative abundance in one metagenomic library caompared to the other two libraries were classified as “diluted (Dil)” gene (open circles) in that library. Table S1. Summary of chemolithoautotrophic MAGs obtained from the white vent metagenomes. Table S2. Isolation source, growth temperature and pH condition, and optimum conditions of reference genomes.