Supplement of Biogeosciences, 11, 4665–4678, 2014 http://www.biogeosciences.net/11/4665/2014/ doi:10.5194/bg-11-4665-2014-supplement © Author(s) 2014. CC Attribution 3.0 License.
Supplement of Substrate quality alters the microbial mineralization of added substrate and soil organic carbon
S. Jagadamma et al.
Correspondence to: S. Jagadamma ([email protected]) Supplement material
Table S1. Quantitative polymerase chain reaction (qPCR) conditions and primers.
Experimental details Bacteria Fungi Primers Eub 338*, Eub nu-SSU-1196F‡, 518† nu-SSU-1536R§ Reaction conditions Denaturation 95 °C, 30sec 95 °C, 30sec Annealing 53 °C, 30 sec 56 °C, 30 sec Elongation 72 °C, 1min 72 °C, 1min Cycle length 39 39 Reaction mixture Sample 1 µL 1 µL Super mix 10 µL 10 µL Water 5 µL 5 µL Primers 2 µL per primer 2 µL per primer Pure cultures for Escherichia coli Saccharomyces standard curves cerevisiae
*5′-ACTCCTACGGGAGGCAGCAG-3′ (Lane, 1991) †5′-ATTACCGCGGCTGCTGG-3′ (Muyzer et al., 1993) ‡5′-GGAAACTCACCAGGTCCAGA-3′ (Borneman & Hartin, 2000) §5′-ATTGCAATGCYCTATCCCCA-3′ (Borneman & Hartin, 2000)
Table S2. Cumulative soil organic C (SOC) respiration after 270 days.
SOC respiration Soils Substrates mg C g-1 soil % of initial C lost Mollisol Unamended 4.1±0.21a 13.9 Glucose 4.0±0.07a 13.3 Starch 3.9±0.15a 13.1 Cinnamic acid 4.0±0.18a 13.6 Stearic acid 4.3±0.12a 14.4
Ultisol Unamended 3.5±0.13 b 15.3 Glucose 3.6±0.15b 15.6 Starch 4.0±0.10ab 17.4 Cinnamic acid 4.4±0.23a 19.0 Stearic acid 4.5±0.04a 19.6
Andisol Unamended 3.4±0.15 b 5.3 Glucose 3.5±0.04b 5.5 Starch 3.6±0.11b 5.5 Cinnamic acid 4.6±0.21a 7.2 Stearic acid 4.4±0.04a 6.8
Gelisol Unamended 2.4± 0.6b 11.8 Glucose 2.7±0.17b 12.9 Starch 2.6±0.0b 12.6 Cinnamic acid 2.9±0.11ab 14.2 Stearic acid 3.1±0.11a 15.3 Each value represents mean ± standard error (n=3). Different letters indicate significant mean differences among substrate addition treatments in each soil.
3000
)
-1
2500
2000
1500
1000
Toluene 500 Water Ethanol
Cumulative SOC respirationCumulative (ug C g soil 0 0 10 20 30 40 50
Incubation time (days) Fig. S1. Comparison of the effect of three solvents (water, ethanol and toluene) on soil organic carbon respiration from Andisol. Symbols represent cumulative soil organic carbon respiration in each sampling time along with standard error bar (n=3). This preliminary experiment demonstrated that the organic solvents ethanol and toluene did not alter the microbial respiration of soil organic carbon.
1e+11 1e+11 Unamended B 1e+10 A Glucose 1e+10
scale) scale) Starch
a 10 1e+9 a a 10 1e+9 a ab b Cinnamic acid b b 1e+8 c a a a Stearic acid 1e+8 c a aba b b a a a a a c b ab b a a a 1e+7 c 1e+7 1e+6 1e+6
1e+5 1e+5
1e+4 1e+4
1e+3 1e+3
1e+2 1e+2
1e+1 1e+1
Bacterial gene copy numbers (log numbers copy gene Bacterial Bacterial gene copy numbers (log gene copy Bacterial 1e+0 1e+0 Day 4 Day 30 Day 270 Day 4 Day 30 Day 270 Incubation length (days) Incubation length (days)
1e+11 1e+11 1e+10 C 1e+10 D
scale) b a a scale) a
10 1e+9 10 1e+9 c b 1e+8 d a 1e+8 c c b a b b b ab a ab b b b a b b b b 1e+7 1e+7 c c c c 1e+6 1e+6
1e+5 1e+5
1e+4 1e+4
1e+3 1e+3
1e+2 1e+2
1e+1 1e+1
Bacterial gene copy numbers (log numbers copy gene Bacterial
1e+0 (log numbers copy gene Bacterial 1e+0 Day 4 Day 30 Day 270 Day 4 Day 30 Day 270 Incubation length (days) Incubation length (days) Fig. S2. Bacterial gene copy numbers in response to the addition of substrates in Mollisol (A), Ultisol (B), Andisol (C), and Gelisol (D).at three time periods during the incubation. Bars represent mean ± standard error calculated from three analytical replications expressed in dry soil mass basis.
1e+12 1e+11 a Unamended B a a 1e+11 A Glucose 1e+10 a ab b b c Starch scale) c 1e+10 1e+9 c d scale) Cinnamic acid
10
10 1e+9 a Stearic acid a a ab a 1e+8 b 1e+8 c b b abc b b c 1e+7 1e+7 c c c b 1e+6 1e+6 c c c 1e+5 1e+5 1e+4 1e+4 1e+3 1e+3 1e+2 1e+2 1e+1
Fungal gene copy numbers (log numbers copy gene Fungal 1e+1
1e+0 (log numbers copy Fungal gene 1e+0 Day 4 Day 30 Day 270 Day 4 Day 30 Day 270 Incubation length (days) Incubation length (days)
1e+11 1e+11 1e+10 C 1e+10 D
scale) 1e+9 scale) 1e+9 10 a 10 a a 1e+8 c b 1e+8 b c c b c 1e+7 d 1e+7 a b b 1e+6 1e+6 d d e a d c a e 1e+5 ab b ab b 1e+5 b 1e+4 1e+4 b c 1e+3 1e+3 d 1e+2 1e+2
1e+1 1e+1
Fungal gene copy numbers (log numbers copy Fungal gene 1e+0 (log numbers copy Fungal gene 1e+0 Day 4 Day 30 Day 270 Day 4 Day 30 Day 270 Incubation length (days) Incubation length (days) Fig. S3. Fungal gene copy numbers in response to the addition of substrates in Mollisol (A), Ultisol (B), Andisol (C), and Gelisol (D). at three time periods during the incubation. Bars represent mean ± standard error calculated from three analytical replications expressed in dry soil mass basis.
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