1 Supplementary Information 2 3 Resuscitation of soil microbiota after > 70-year of desiccation 4 Jun Zhao1, Dongfeng Chen1*, Wei Gao1,2, Zhiying Guo1,2, Zhongjun Jia1, Marcela 5 Hernández3* 6 1State Key Laboratory of Soil and Sustainable Agriculture. Institute of Soil Science, 7 Chinese Academy of Sciences. Nanjing, 210008, Jiangsu Province, China 8 2University of Chinese Academy of Sciences, Beijing 100049, China 9 3School of Environmental Sciences, Norwich Research Park, University of East 10 Anglia, Norwich, UK 11 * Correspondence: 12 Dongfeng Chen: [email protected] 13 Marcela Hernández: [email protected] 14 Running title: Soil microbes after long-term desiccation 15 1 16 17 18 Supplementary Figure 1. The geographical distribution of soils collected between 19 1934-1939. The soils were collected from different districts across mainland of China. 20 2 21 22 Supplementary Figure 2. Changes in bacterial abundance in 24 soils. The bacterial 23 abundance was estimated at desiccated condition (Dry) and after rewetting incubation 24 (Rewet) based on real-time quantitative PCR of 16S rRNA genes and high-throughput 25 sequencing, and shown as mean value of 2-3 biological replicates with standard 26 errors. 3 A. Dry 0 0.5 1 1.5 2 Counts S14 S16 S13 S15 S21 S19 S04 S05 S10 S11 S24 S02 S08 S20 S07 S18 S01 S12 S06 S09 S17 S03 S22 S23 Pseudonocardiaceae (OTU-2) Pseudonocardiaceae (OTU-24) Actino- Pseudonocardiaceae (OTU-25) bacteria Pseudonocardiaceae (OTU-14) Nocardiaceae (OTU-11) Anaerolineae (OTU-104) Chloroflexi SIA-15 (OTU-78) Stenotrophomonas (OTU-44) Proteobacteria Planococcaceae (OTU-1) Tumebacillus (OTU-13) Tumebacillus (OTU-19) Bacillaceae (OTU-9) Paenibacillus (OTU-15) Bacillales (OTU-3) Saccharopolyspora (OTU-59) Paenibacillaceae (OTU-22) Bacillales (OTU-4) Paenibacillaceae (OTU-72) Paenibacillus (OTU-54) Paenibacillaceae (OTU-83) Paenibacillus (OTU-64) Cohnella (OTU-102) Firmicutes Clostridium (OTU-60) Selenomodales (OTU-29) Hydrogenispora (OTU-49) Clostridium (OTU-48) Pelotomaculum (OTU-66) Hydrogenispora (OTU-142) Peptococcaceae (OTU-52) Clostridium (OTU-173) Hydrogenispora (OTU-304) Clostridium (OTU-88) Desulfosporosinus (OTU-196) Hydrogenispora (OTU-474) Clostridium (OTU-101) Ruminococcaceae (OTU-139) 27 28 Supplementary Figure 3. Heatmap of the most relevant OTUs derived from total 29 Bacteria. A. Dry soils; B. rewetted soils. The colored scale gives the percentage 30 abundance of OTUs. 31 4 B. Rewetted 0 1 2 3 Counts S22 S23 S08 S09 S01 S02 S03 S12 S18 S06 S07 S20 S19 S21 S10 S11 S24 S04 S05 S14 S17 S15 S13 S16 Micrmonosporaceae (OTU-55) Actino- Nocardiaceae (OTU-11) bacteria Halomonas (OTU-36) Aminobacter (OTU-90) Proteo- Acinetobacter (OTU-85) bacteria Stenotrophomonas (OTU-44) Ralstonia (OTU-87) Alicyclobacillus (OTU-6) Alicyclobacillus (OTU-20) Alicyclobacillus (OTU-8) Planococcaceae (OTU-1) Tumebacillus (OTU-13) Cohnella (OTU-30) Brevibacillus (OTU-23) Bacillaceae (OTU-9) Bacillus (OTU-230) Sporolactobacillaceae (OTU-5) Bacillales (OTU-4) Bacillales (OTU-3) Paenibacillus (OTU-15) Firmicutes Paenibacillaceae (OTU-22) Clostridiales (OTU-105) Bacillales (OTU-45) Bacillaceae (OTU-32) Ammoniphilus (OTU-41) Alicyclobacillus (OTU-124) Alicyclobacillus (OTU-527) Pullulanibacillus (OTU-31) Selenomodales (OTU-29) Hydrogenispora(OTU-69) Clostridium (OTU-60) Paenibacillus (OTU-54) Paenibacillaceae (OTU-72) 32 33 Supplementary Figure 3. continues 34 5 35 36 Supplementary Figure 4. Composition of the archaea in soils at desiccated condition 37 (Dry) and after rewet incubation (Rewet). The archaeal composition was plotted at 38 class level and the most abundant classes were shown. Taxa not seen more than 3 39 times in at least 20 % of the samples were removed using phyloseq package on R. The 40 relative abundances of minor classes were summed up and shown as “others” which 41 do not exceed 6.8% of total archaeal reads in any given soil. 6 .