1
Supplementary material
2
Journal: Metabolomics
3
4
Impact of induced drought stress on the metabolite
5
profiles of barley grain
6
7
Alexandra Wenzel1, Thomas Frank1, Gabriela Reichenberger2, Markus Herz2 and
8
Karl-Heinz Engel*1
9
1Technische
10
Universität München,
11
Lehrstuhl für Allgemeine Lebensmitteltechnologie,
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Maximus-von-Imhof-Forum 2, D-85350 Freising-Weihenstephan, Germany
13
14
2Bavarian
State Research Center for Agriculture (LfL), Institute for Crop Science and
Plant Breeding, Am Gereuth 8, D-85354 Freising, Germany
15
16
17
* Author to whom correspondence should be addressed [Karl-Heinz Engel,
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Technische Universität München, Lehrstuhl für Allgemeine Lebensmitteltechnologie,
19
Maximus-von-Imhof-Forum
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Telephone: +49 (0) 8161 71 4250; Fax: +49 (0) 8161 71 4259; email:
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k.h.engel@wzw.tum.de]
2,
D-85350
22
1
Freising-Weihenstephan,
Germany;
23
Supplementary Table S1 Analyzed spring barley genotypes and their
24
characteristics
Barke
IPZ 24727
Argentina
Mutant 6519
Steina
Streif
Polygena
Supplier
Josef Breun GmbH & Co. KG,
Herzogenaurach Germany
Bavarian State Research Center for
Agriculture (LfL), Germany
Cerveceria y Malteria Quilmes,
Argentina
Josef Breun GmbH & Co. KG,
Herzogenaurach Germany
Origin
Libelle x Alexis
(1996)
Maresi x Omega
Streng-Engelen GmbH & Co.KG,
Uffenheim, Germany
Saatzucht Hadmersleben GmbH,
Hadmersleben, Germany
Pasadena x Aspen
(2007)
46401-80 x 45465-78
(1994)
25
26
2
chemical-induced mutant of
Quilmes Paine (1993)
(Sultan x St.434/62) x (Voldagsen
x Carlsberg x Volla) (1980)
27
Supplementary Table S2 Compounds identified in fraction I (major lipids) and
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fraction II (minor lipids)
no. compound
identa
saturated FAMEb
2
3
5
9
12
16
18
21
23
26
29
compound
identa
unsaturated FAMEb
12:0
14:0
15:0
16:0
17:0
18:0
19:0
20:0
21:0
22:0
23:0
A
A
A
A
A
A
D
A
A
A
D
7
8
11
15
20
25
28
30
6
13
19
31 24:0
A
14
35 26:0
37 28:0
D
D
24
free fatty acidsc
1 12:0
2 14:0
3 15:0
5 16:1 (9E)
6 16:1 (9Z)
7 16:0
8 17:1 (9Z)
9 17:0
12 18:2 (9Z, 12Z)
13 18:3 (9Z, 12Z, 15Z)
14 18:1 (9Z)
15 18:0
17 20:2 (11Z, 14Z)
18 20:1 (11Z)
19 20:0
21 22:1 (13Z)
22 22:0
24 24:0
29
30
31
32
33
34
35
36
37
no.
A
A
A
C
A
A
A
A
A
A
A
A
A
A
A
A
A
A
16:1 (9E)
16:1 (9Z)
17:1 (9Z)
18:1 (9Z)
20:1 (11Z)
22:1 (11Z)
23:1
24:1 (15Z)
16:2 (9Z, 12Z)
18:2 (9Z, 12Z)
20:2 (11Z, 14Z)
18:3 (9Z, 12Z,
15Z)
22:2 (13Z,16Z)
fatty alcoholsc
4
16:0
10 18:0
11 phytol
16 20:0
20 22:0
23 24:0
25 26:0
27 28:0
35 30:0
tocopherolsc
26 -tocopherol
a Identification
no. compound
hydrocarbons
D
A
A
A
A
A
D
A
B,D
A
A
1
4
10
17
22
27
32
33
34
36
37
C14
C18
C20
C22
C24
C26
C28
squalene
cholestane
C31
C32
A
A
A
A
A
A
A
A
A
D
D
A
D
A
A
A
A
A
C
A
D
D
B,C
sterols/stanolsc
28 cholesterol
29 campesterol
30 campestanol
31 stigmasterol
32 β-sitosterol
33 sitostanol
34 5-avenasterol
36 gramisterol
37 cycloartenol
38 7-avenasterol
39 24-methylenecycloartenol
40 citrostadienol
according to
A mass spectral data and retention times of reference compounds
B mass spectral data and retention index of Golm Metabolome Database (Kopka et al., 2005)
C mass spectral data of NIST08 mass spectral library
D mass spectral data
E according to (Xu and Godber, 1999)
F according to (Kamal-Eldin et al., 1992)
b fatty acid methyl esters
c TMS derivatives of respective compound
3
identa
A
A
A
A
A
A
F
F
E
F
A
F
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Supplementary Table S3 TMS derivatives of compounds identified in fraction III
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(sugars and sugar alcohols) and fraction IV (acids, amino
40
41
acids and amines)
no.
42
43
44
45
46
47
48
a
b
compounda
identb
fraction
sugars III
and sugar alcohols
1,2
arabinose
ad
3
xylitol
4,5,6 fructose
7,9
galactose
8,12 glucose
10
mannitol
11
sorbitol
myo-inositol
13
14
sucrose
15,16 maltose
17
galactinol
18
raffinose
A
A
A
A
A
A
A
A
A
A
A
A
##
acids
lactic acid
1
lactic acid
3
pyruvic acid
A
6
benzoic acid
9
phosphoric acid
13
maleic acid
14
succinic acid
15
glyceric acid
16
fumaric acid
19
glutaric acid
22
malic acid
27
threonic acid
α-ketoglutaric acid
30
33
3-glycerophosphoric acid
34
cis-aconitic acid
36
2-glycerophosphoric acid
38
citric acid
39
isocitric acid
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
C
compounda
identb
amino acids and amines
2
alanine
4
α-aminobutyric acid
5
valine
7
ethanolamine
8
leucine
10
proline
11
isoleucine
12
glycine
17
serine
18
threonine
β-alanine
20
21
homoserine
23
pyroglutamic acid
24
-aminobutyric acid
25
aspartic acid
26
cysteine
28
phenylalanine
29
glutamic acid
31
asparagine
32
putrescine
35
glutamine
37
ornithine
40
lysine
41
tyrosine
42
tryptophan
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
D
A
A
A
A
B,D
A
A
A
A
no.
Metabolites identified as persilylated derivates
Identification according to
A mass spectral data and retention times of reference compound
B mass spectral data and retention index of Golm Metabolome Database (Kopka et al., 2005)
C NIST08 mass spectral library
D mass spectral data
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50
4
51
Captions
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53
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55
56
57
58
59
60
61
62
Supplementary Fig. S1
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Supplementary Fig. S3
65
66
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Rain-Out-Shelter (ROS)
Supplementary Fig. S2
Soil moisture content (,,) and irrigation/natural rainfall (bars)
under drought stress in a Rain-Out-Shelter (a) and under natural
weather conditions at control location 1 (b) in growing seasons 2010
(, black bars), 2011(, grey bars) and 2012(, broken bars)
Hours of sun recorded at Munich airport (approximately 0.5 kilometres
from C1/ROS and C2) during growing seasons 2010, 2011 and 2012.
Graphics provided on http://weather-online.de (23 June 2014)
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68
Supplementary Fig. S1
69
70
6
10-May
10.5
17-May
17.5
24-May
24.5
31-May
31.5
*
*
7-Jun
7.6
14-Jun
14.6
21-Jun
21.6
28-Jun
28.6
5-Jul
5.7
12-Jul
12.7
19-Jul
19.7
26-Jul
26.7
25
20
15
10
5
0
60
50
40
30
20
10
0
irrigation [mm]
rainfall [mm]
0
3.5
-100
3-May
a
26.4
-200
26-Apr
-300
19.4
-400
19-Apr
-500
12.4
-600
12-Apr
-700
5.4
-800
-900
5-Apr
* continuing data
irrigation 20102010
Bewässerung
irrigation 20112011
Bewässerung
irrigation 20122012
Bewässerung
soil moisture tension 2010
2010
soil moisture tension 2011
2011
soil moisture tension 2012
2012
rainfall 2010 2010
Niederschlag
rainfall 2011 2011
Niederschlag
rainfall 2012 2012
Niederschlag
soil moisture tension 2012
2012
soil moisture tension 2011
2011
soil moisture tension 2010
2010
7
b
0
-100
-200
-300
-400
-500
-600
-700
-800
-900
29.3
a data of tensiometer obtained in 60 cm soil depth
not available due to technical problems
29-Mar
soil moisture tension [mbar]a
soil moisture tension [mbar]a
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
Supplementary Fig. S2
71
Supplementary Fig. S3
122
123
124
16 March – 31 August 2010
sun hours per day
121
125
126
127
128
129
sun hours per day
16 March – 31 August 2011
130
131
132
133
134
sun hours per day
16 March – 31 August 2012
135
8