Plant and Soil
Jaiswal et al.
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Supplementary Material
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GC/MS analysis of aqueous biochar extracts
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Aqueous extracts of the biochars were prepared by shaking 1 g biochar in 10 ml
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deionized water (hereafter, water) in 15 ml polypropylene centrifuge tubes in the dark for
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24 hours, followed by sedimentation and filtration of the liquid solution via membrane
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filters of 0.22-µm pore size (Durapore PVDF membrane, Millipore Corp., Carrigtwohill,
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Ireland). Aliquots (2 ml) of the extracts were lyophilized and the dried residue subjected
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to a two-stage derivatization procedure immediately prior to analysis by GC/MS
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following the methods detailed in Graber et al. (2014). The residue was first
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methoxyamininated to stabilize carbonyl moieties, and then functional groups with active
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hydrogens were silylized with N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA).
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The derivatized extracts were analyzed by quadrupole GC/MS (Agilent Technologies,
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Santa Clara, California, USA) using electron impact ionization. Separation was achieved
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on a 30-m long, VF-5M, 0.25 µm film, capillary column (Varian Inc., Palo Alto,
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California, USA). Total ion chromatograms (TIC) were analyzed with freely available
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deconvolution software (Automated Mass Spectral Deconvolution and Identification
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System; AMDIS) at a minimum match probability of 75% and retention indices based on
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a series of alkanes. The deconvoluted mass spectra were compared with a specialty
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library for plant, animal and microorganism metabolites (Golm Metabolite Database)
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freely provided by the Max Planck Institute for Metabolic Plant Physiology (Golm,
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Germany) and to the NIST08 mass spectral library. Results given in Table 5,
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Supplementary Material.
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Plant and Soil
Jaiswal et al.
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Table 1 Analysis of variance (ANOVA) to determine the effect of interaction between
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pyrolysis temperature (HTT) and concentration of eucalyptus (EUC) and greenhouse
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pepper plant wastes (GHW) biochar on disease caused by R. solani in bean.
Source
EUC
GHW
AUMPCa
Disease
AUMPCa Disease
(%×days)
severity (%)b (%×days) severity (%)b
Model
0.0028*
<0.0001*
<0.0001*
<0.0001*
R2
0.3047
0.4687
0.4047
0.4075
0.0870
0.2148
0.7078
0.1504
Concentration (C ) 0.0078*
<0.0001*
0.0015*
<0.0001*
T×C
0.0590
0.7327
0.2239
ANOVA
P-value
Temperature (T)
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a
27
b
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*
0.3666
Area under mortality progress curve
Values were normalized by an arcsine square root transformation before analysis.
Significant at α < 0.05
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31
32
33
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Plant and Soil
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Table 2 Analysis of variance (ANOVA) to determine the effect of interaction between
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feedstocks and concentration of biochar on disease caused by R. solani in bean.
AUMPCa (%×days)
Disease severity (%)b
Model
0.0057*
0.0006*
R2
0.2566
0.3233
Feedstock (F)
0.8654
0.4740
Concentration (C )
0.0224*
0.0244*
F×C
0.0084*
0.0005*
Source
ANOVA
P-value
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a
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b
39
*
Area under mortality progress curve
Values were normalized by an arcsine square root transformation before analysis
Significant at α < 0.05
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42
43
44
45
46
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Table 3 Analysis of variance (ANOVA) to determine the effect of interaction between R.
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solani inoculation (inoculated and non-inoculated), pyrolysis temperature (HTT) and
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concentration of eucalyptus (EUC) and greenhouse pepper plant wastes (GHW) biochar
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on growth parameters of bean.
Source
EUC
GHW
Plant height
Shoot dry
Plant height
Shoot dry
(cm)
weight (g)
(cm)
weight (g)
Model
<0.0001*
<0.0001*
<0.0001*
<0.0001*
R2
0.7297
0.7999
0.7108
0.6757
<0.0001*
<0.0001*
<0.0001*
<0.0001*
Temperature (T )
0.0636
0.1025
0.0002*
0.0090*
Concentration (C )
0.5377
0.0362*
0.0049*
0.4418
I×T
0.2140
0.5891
0.0003*
0.0034*
I×C
0.0820
0.0239*
0.7185
0.0759
T×C
0.8353
0.9835
<0.0001*
<0.0001*
I×T×C
0.8892
0.3383
0.0139*
0.0864
ANOVA
P-value
Rhizoctonia inoculation
(I)
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Significant at α < 0.05
*
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Jaiswal et al.
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Table 4 Analysis of variance (ANOVA) to determine the effect of pyrolysis temperature
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(HTT) and concentration of eucalyptus (EUC) biochar on growth parameters of bean
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(non-inoculated).
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Source
EUC
GHW
Plant height
Shoot dry
Plant height
Shoot dry
(cm)
weight (g)
(cm)
weight (g)
Model
<0.0001*
<0.0001*
<0.0001*
<0.0001*
R2
0.8055
0.7967
0.8095
0.7057
Temperature (T )
0.0111*
0.1570
<0.0001*
<0.0001*
Concentration (C )
0.0574
0.0117*
0.0068*
0.5742
T×C
0.8499
0.6069
<0.0001*
<0.0001*
ANOVA
P-value
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Significant at α < 0.05
*
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Table 5 Putative identifications by gas chromatograph/mass spectrometer (GC/MS) of
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compounds in aqueous extracts of eucalyptus woodchips (EUC) biochars produced at
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350◦C (EUC-350) and 600◦C (EUC-600) and greenhouse pepper plant waste (GHW)
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biochars produced at 350◦C (GHW-350) and 600◦C (GHW-600).
Retention EUC-
EUC-
GHW-
GHW-
Compound
time
350
600
350
600
2-Hydroxypropanoic acid
18.379
Y
Y
Y
Lactic acid
18.396
Y
Y
Y
Hexanoic acid
19.297
Y
Hydroxyacetic acid (glycolic acid) (alpha-hydroxy acid)
19.542
Y
1,2-Butanediol
22.163
Y
3-Hydroxypropanoic acid
23.074
Y
Heptanoic acid
24.102
Y
Y
4-Hydroxybutanoic acid
26.660
Y
Y
Benzoic acid
26.920
Y
Y
Octanoic acid
27.598
Glycerol
27.898
Succinic acid
29.189
3-Methylbenzoic acid
29.333
2-Methylbutanedioic acid
29.516
Glyceric acid
29.654
2-Methylbenzoic acid
30.216
Y
Nonanoic acid
30.554
Y
Oxalic acid
31.522
Y
Glutaric acid
31.831
Y
Benzenepropanoic acid
32.120
Y
Sulfurous acid, 2-ethylhexyl isohexyl ester
33.982
Decanoic acid
33.208
Y
Salicylic acid
34.432
Y
1,4,5-Pentatriol
34.462
Pyroglutamic acid
34.765
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
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Plant and Soil
Jaiswal et al.
2-Piperidinecarboxylic acid
34.825
Y
Erythronic acid
35.297
Y
Threonic acid
35.299
Y
4-Hydroxybenzoic acid
35.887
Y
3-hydroxybenzoic acid
37.344
Y
Methyl α-d-lyxofuranoside
38.043
Y
D-Altro-2-Heptulose (sedoheptulose)
38.307
Y
1,6-Anhydro-beta-d-glucose
38.520
Y
1,6-Anhydroglucose
38.986
Y
Cyperaquinone
39.205
4-Benzylimidazole-5-(1-propenoic acid), methyl ester
39.274
Y
Tetradecanoic acid
42.248
Y
Mannitol
43.613
Y
Silane, dimethyl(4-(2-phenylprop-2-yl)phenoxy)butoxy-
43.748
Hexadecanoic acid
46.035
Myo-Inositol
46.618
Octadecanoic acid
49.573
Trehalose
57.061
Melezitose
57.130
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
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References
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Graber ER, Tsechansky L, Lew B, Cohen E (2014) Reducing capacity of water extracts
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of biochars and their solubilization of soil Mn and Fe. Eur J Soil Sci 65:162-172.
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