Cover sheet for SI
Authors:
Li-Lian Wen, Yin Zhang, Ya-Wei Pan, Wen-Qi Wu, Shao-Hua Meng, Chen Zhou, Youneng Tang,Ping Zheng,He-Ping Zhao*
Manuscript title: The Roles of Methanogens and Acetogens in Dechlorination of Trichloroethene Using Different Electron Donors.
Number of pages: 9
Number of table: 5
Number of figures: 2
Supplementary information (SI) for manuscript
The Roles of Methanogens and Acetogens in Dechlorination of Trichloroethene Using Different Electron Donors
Li-Lian Wen1, YinZhang1, Ya-Wei Pan1, Wen-Qi Wu1, Shao-Hua Meng1, Chen Zhou2, Youneng Tang3, PingZheng1, He-Ping Zhao1*
1. MOE Key Lab of Environmental Remediation and Ecosystem Health, College of Environmental and Resource Sciences, Zhejiang University, Hangzhou, China.
2. Department of Civil and Environmental Engineering, FAMU-FSU College of Engineering, Florida State University, Tallahassee, Florida, 32310-6046.
* Correspondance to Dr. He-Ping Zhao. Tel: 0086-571-88982739, Fax: 0086-571-88982739, E-mail: ;
Table S1. Primers and PCR conditions for tested genes
95 oC 2 min (95oC 10 sec 56 oC 20 sec 68 oC 20 sec)×40 72oC 1 min / 16SF 16SR / 5'-GTGSTGCAYGGYTGTCGTCA-3' 5'-ACGTCRTCCMCACCTTCCTC-3' / 16S rDNAfor bacteria / Maeda et al., 2003 / -3.215 / 1.05
94 oC 10 min (94oC 30 sec 58 oC 20 sec 72 oC 60 sec)×40 72oC 1 min / ARC787F
ARC1059R / 5'-ATTAGATACCCSBGTAGTCC-3'
5'-GCCATGCACC WCCTCT-3' / 16S rDNAfor archaea / Yu et al., 2005 / -3.630 / 0.89
94 oC 10 min (94oC 30 sec 58 oC 300 sec 72 oC 60 sec)×40 72oC 1 min / Mlas
rev / 5'-GGTGGTGTMGGDTTCACMCARTA-3'
5'-CGTTCATBGCGTAGTTVGGRTAGT-3' / mcrA / Steinberg et al., 2008 / -3.221 / 1.04
95 oC 2 min (95oC 30 sec 58 oC 30 sec 72 oC 20 sec)×30 72oC 10 min / fhs1
FTHFS-r / 5'-GTWTGGGCWAARGGYGGMGAAGG-3'
5'-GTATTGDGTYTTRGCCATACA-3' / FTHFS / Kewei et al., 2009 / -3.442 / 0.95
95 oC 2 min (95oC 30 sec 58 oC 30 sec 72 oC 10 sec)×30 72oC 10 min / Dhc1200F
Dhc1271R / 5'-CTGGAGCTAATCCCCAAAGCT-3'
5'-CAACTTCATGCAGGCGGG-3' / DHC / He et al., 2003 / -3.290 / 1.01
Table S2. Stoichiometric equations of the dominant biochemical reactions in YCQ 1 culture with different electron donors
Lactate: 1) H3CH2OCOO-+2H2O=CH3COO-+HCO3-+2H2+H+2) CH3CH2OCOO-+H2=CH3CH2COO-+H2O
3) HCO3-+2H2+1/2H+= 1/2CH3COO-+2H2O
4) HCO3- +4H2+H+=CH4+3H2O
Acetate: 1) CH3COO- + 2C2HCl3+ 4H2O =2HCO3- + 4Cl- + 2C2H3Cl +5H+
2) HCO3- +4H2+H+=CH4+3H2O
Methanol: 1) CH3OH+2H2O=3H2+HCO3-+H+
2) CH3OH+1/2CO2= 3/4CH3COOH+2H2O
3) HCO3-+2H2+H+=1/2CH3COOH+2H2O
4) HCO3- +4H2+H+=CH4+3H2O
5) 4CH3OH=3CH4+CO2+2H2O
Hydrogen:1) HCO3-+2H2+H+=1/2CH3COOH+2H2O
2) HCO3- +4H2+H+=CH4+3H2O
Reaction for dechlorination for all electron donors except acetate:
C2HCl3+3H2=C2H4+3Cl-+3H+
Table S3. Electron-donor distribution for biomass synthesis based on method of Ziv-El et al.(2012)
Steady state endProduct (mM) / Microbial process
(Table S2) / Microorganism / Steady state biomass () / fs0
propionate / Fermenter / / 0.18a
acetate / Fermenter or Homoacetogen / / 0.18
methane / Methanogen / / 0.05b
Chlorinated ethenes / Dehalococcoides / / 0.10c
is the acetate used by Dehalococcoides as the carbon source. Biomass is C5H7NO2, and N is from NH4+, which gives 20 me- eq/mmol biomass (Rittmann & McCarty, 2001).
a: Rittmann & McCarty, 2001.
b: Rittmann & McCarty, 2001. Table 3.1
c: Ziv-El et al., 2012.
Table S4. Pearson correlation analysis of dechlorination rates with methane concentration, mcrA,Dhc, and mcrA/Dhc ratios (Batch Test 2).
Methane / mcrA / Dhc / Dhc/Mcr / k1 / k2 / k3Methane (85-420M) / Pearson / 1 / 0.791 / -0.750 / -0.991 / -0.152 / -0.990 / -0.988
Sig. 2-tailed / n.a. / 0.419 / 0.460 / 0.086 / 0.903 / 0.091 / 0.099
mcrA / Pearson / 0.791 / 1 / -0.998* / -0.701 / -0.725 / -0.869 / -0.686
Sig. 2-tailed / 0.419 / n.a. / 0.040 / 0.505 / 0.484 / 0.329 / 0.518
Dhc / Pearson / -0.750 / -0.998* / 1 / 0.655 / 0.767 / 0.836 / 0.639
Sig. 2-tailed / 0.460 / 0.040 / n.a. / 0.546 / 0.443 / 0.369 / 0.559
Dhc/Mcr / Pearson / -0.991 / -0.701 / 0.655 / 1 / 0.018 / 0.962 / 1.000*
Sig. 2-tailed / 0.086 / 0.505 / 0.546 / n.a. / 0.989 / 0.176 / 0.013
k1 / Pearson / -0.152 / -0.725 / 0.767 / 0.018 / 1 / 0.290 / -0.003
Sig. 2-tailed / 0.903 / 0.484 / 0.443 / 0.989 / n.a. / 0.813 / 0.998
k2 / Pearson / -0.990 / -0.869 / 0.836 / 0.962 / 0.290 / 1 / 0.956
Sig. 2-tailed / 0.091 / 0.329 / 0.369 / 0.176 / 0.813 / n.a. / 0.189
k3 / Pearson / -0.988 / -0.686 / 0.639 / 1.000* / -0.003 / 0.956 / 1
Sig. 2-tailed / 0.099 / 0.518 / 0.559 / 0.013 / 0.998 / 0.189 / n.a.
* Significant correlation (P<0.05). **Significant correlation (P<0.01). n.a. means not applicable.
k1: TCE to cis-DCE, k2: cis-DCE to VC; k3: VC to ethene.
Table S5. Pearson correlation analysis of dechlorination rates with methane concentration, mcrA,Dhc, and mcrA/Dhc ratios (Batch Test 3).
Methane (0-4 mM) / mcrA / Dhc / Dhc/mcrA / k1 / k2 / k3Methane (0-4 M) / Pearson / 1 / 0.681 / 0.472 / -0.551 / 0.691 / 0.940** / 0.854*
Sig. 2-tailed / 0.136 / 0.344 / 0.257 / 0.129 / 0.005 / 0.031
mcrA / Pearson / 0.681 / 1 / 0.837* / -0.695 / 0.938** / 0.835* / 0.950**
Sig. 2-tailed / 0.136 / 0.037 / 0.125 / 0.006 / 0.039 / 0.004
Dhc / Pearson / 0.472 / 0.837* / 1 / -0.203 / 0.601 / 0.702 / 0.725
Sig. 2-tailed / 0.344 / 0.037 / 0.700 / 0.207 / 0.120 / 0.103
Dhc/mcrA / Pearson / -0.551 / -0.695 / -0.203 / 1 / -0.881 / -0.522 / -0.730
Sig. 2-tailed / 0.257 / 0.125 / 0.700 / 0.021 / 0.288 / 0.100
k1 / Pearson / 0.691 / 0.938** / 0.601 / -0.881 / 1 / 0.773 / 0.927**
Sig. 2-tailed / 0.129 / 0.006 / 0.207 / 0.021 / 0.071 / 0.008
k2 / Pearson / 0.940** / 0.835* / 0.702 / -0.522 / 0.773 / 1 / 0.944**
Sig. 2-tailed / 0.005 / 0.039 / 0.120 / 0.288 / 0.071 / 0.005
k3 / Pearson / 0.854* / 0.950** / 0.725 / -0.730 / 0.927** / 0.944** / 1
Sig. 2-tailed / 0.031 / 0.004 / 0.103 / 0.100 / 0.008 / .005
* Significant correlation (P<0.05). **Significant correlation (P<0.01).
k1: TCE to cis-DCE, k2: cis-DCE to VC; k3: VC to ethene.
Figure S1. Time course of dechlorination of TCE for Batch Test 1 using different electron donors.
Figure S2. Theabsolute abundances of bacterial groups at the genus level for tested electron donors at the end of Batch Test 2.
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