Table S1. Primers used for gene cloning and qPCR in this study.
Name
Sequence (5’-3’) a
Ht1-FEH I-F
5' GATGGTAAAGGAGATGGC 3'
Ht1-FEH I-R
Ht1-FEH II-F
Ht1-FEH II-R
5' TGAAGCAGGTGTCAAAAC 3'
5' CTTAAAACTCACGGATACC 3'
5' AAAGCAACAAAGAATGACC 3'
Pic-Ht1-FEH I-F
Pic-Ht1-FEH I-R
Pic-Ht1-FEH II-F
Pic-Ht1-FEH II-R
a
Remark
Amplification of
full-length
sequences
5' CCGCTCGAGAAAAGATCAGAAGATTTGCAGCCTT 3' Amplification of the
entire
5' GCCGTCTAGAAAATCCATAGGAACTATTTGAG 3'
Ht1-FEH-ORF for
5' CCGCTCGAGAAAAGAACTGGTCGGAGTATGGTA 3'
expression
5' GCCGTCTAGAAAAGTTGCACTTTTTACG 3'
qPCR Ht1-FEH I-F
5' ACCGATCAGTTTCTCCTTGCTACC 3'
qPCR Ht1-FEH I-R
5' CCATGCCCATAACACCCTTC 3'
qPCR Ht1-FEH II-F
5' ACCCAATAATAACCCCACCCG 3'
qPCR Ht1-FEH II-R
qPCR Ht Actin-F
qPCR Ht Actin-R
5' CGTACCGAGTCCAGGTTACAA 3'
5' ATGTATGTAGCCATCCAGG 3'
5' TGTTAGGTCACGCCCAG 3'
Primers for
gene expression
analysis
Restriction sites are underlined and the Kex2 cleavage site is marked in bold. F and R refer to forward
and reverse primers, respectively.
1
Table S2. LC-MS/MS analysis of peptides after tryptic digestion of Ht1-FEH I obtained from
SDS-PAGE.
Sequences（from N to C terminus)
a
SLVDHSIVESFGGEGLAcITAR
SSIAQGVDKSTYGAFLDLDPLHDK
NVSTSGSYGPFGVLVFASK
RVLWAWVNEGDSPSDAFKK
GGSLLEVPGISGSQADIEVVFSLSNPSDAELINSDLSDPQHLcEQK
ETNMWEcPDFYPVSNIGK
RVLWAWVNEGDSPSDAFK
VLWAWVNEGDSPSDAFK
VLWAWVNEGDSPSDAFKK
GQATALLYYSTDGFNWTR
WSGNPVLTPVnDINTSQFR
WSGNPVLTPVNDINTSQFR
VLWAWVnEGDSPSDAFK
RVLWAWVnEGDSPSDAFK
KQLVQWPVEEIK
KQLVQWPVEEIKK
STYGAFLDLDPLHDK
nVSTSGSYGPFGVLVFASK
EPYDIGGcFTGSSTILHGSKPIILYTAQDADGAQVQNLALPK
ETnmWEcPDFYPVSNIGK
GQATALLYYSTDGFnWTR
ETnMWEcPDFYPVSNIGK
VFKGPNKFQVLMcSDQSR
FQVLmcSDQSR
SVTmLSLNAWNMNK
ETNmWEcPDFYPVSNIGK
WSGNPVLTPVNDInTSQFRDPSTAWK
FQVLMcSDQSR
GWSGLQSFPR
WSGNPVLTPVNDINTSQFRDPSTAWK
GPnKFQVLMcSDQSR
SVTmLSLNAWNmNK
FQVLMcSDqSR
NLTEQTAVFFR
SVTMLSLNAWnMNK
STYGAFLDLDPLHDKISLR
SVTMLSLNAWNMNK
QLVQWPVEEIK
VLWAWVnEGDSPSDAFKK
VFKGPNKFQVLmcSDQSR
EPYDIGGcFTGSSTILHGSKPIILYTAqDADGAQVQNLALPK
nLTEQTAVFFR
SVTmLSLnAWNMNK
QLVQWPVEEIKK
SDPLLKDWIK
TAFHFQPLK
RVLWAWVnEGDSPSDAFKK
GGSLLEVPGISGSQADIEVVFSLSNPSDAELINSDLSDPQHLcEqK
ILIGSEIIK
SSIAQGVDK
SIWLSDTRK
SVTmLSLnAWNmNK
EmAGWVLSFcILLVVnGVGVHASEDLQPYRTAFHFqPLK
DGIDTSFqGNNTMHVLK
DGIDTSFQGNNTmHVLK
LYVFnnGSK
LAIHEHAKLYVFNNGSK
SIWLSDTR
VSFnSHEYYVIGmYDPqTDQFLLATNDFSVSNTQLQYDYGR
a
PSMs b
Xcorr c
34
4
2
2
2
4
4
35
3
2
12
5
1
2
2
7
15
2
24
8
4
6
2
8
3
5
8
5
12
2
3
7
4
2
9
3
4
2
1
1
26
2
6
9
2
2
1
5
17
1
1
5
2
6
1
1
1
2
1
6.54
6.01
5.76
5.76
5.76
5.70
5.38
5.37
5.13
5.07
4.99
4.59
4.54
4.44
4.43
4.42
4.19
4.09
4.03
3.96
3.93
3.91
3.79
3.78
3.72
3.70
3.65
3.56
3.47
3.42
3.39
3.35
3.33
3.31
3.11
3.08
2.99
2.81
2.80
2.71
2.66
2.66
2.61
2.58
2.57
2.54
2.42
2.37
2.22
2.13
2.06
1.92
1.83
1.73
1.63
1.56
1.40
1.36
1.33
Charge
2
3
2
3
3
2
3
2
2
2
2
2
2
3
2
3
2
2
4
2
2
2
3
2
2
2
3
2
2
3
3
3
2
3
2
4
2
2
3
4
4
2
3
3
2
2
4
5
2
2
2
3
4
3
3
2
4
2
4
d
MH+ [Da] e
2318.15239
2577.28708
1916.97954
2205.10544
4852.36484
2186.95537
2077.02170
1920.91582
2049.01348
2063.98564
2146.06499
2145.07109
1921.90007
2078.00150
1496.84856
1624.94181
1691.82817
1917.95439
4446.24038
2203.93804
2064.96587
2187.93974
2141.06229
1386.61540
1624.78691
2202.95317
2931.43210
1370.62187
1134.56633
2930.44461
1767.81681
1640.77860
1371.60369
1325.68402
1609.76665
2161.13051
1608.78850
1368.75652
2049.99643
2157.06264
4447.23940
1326.67034
1625.76688
1496.85093
1214.67937
1088.58904
2206.09340
4853.35584
985.62950
904.47246
1105.60271
1641.76297
4447.23940
1877.86924
1892.87748
1043.50273
1941.03469
977.50615
4844.18862
The matched sequences are listed according to the predicted amino acid sequences and the amino
acids modified are in lowercase letters.
b
The total number of identified peptide sequences (PSMs = peptide spectrum matches) for the protein
including those redundantly identified.
c and d
All peptide Xcorr values are above the specified score threshold (the minimum score is 1.3 for a
charge state of 2; the minimum score is 1.305 for a charge state of 3; and the minimum score is 1.31 for
2
a charge state of 4).
e
MH+ [Da] (calculated m/z of the peptide with z = 1).
3
Table S3. LC-MS/MS analysis of peptides after tryptic digestion of Ht1-FEH II obtained from
SDS-PAGE.
Sequences (from N to C terminus)
a
PSMs b
Xcorr c
1
4
3
9
2
3
5
2
2
3
1
4
4
3
3
1
1
6
2
3
6
2
1
12
1
2
6
4
4
2
1
3
1
3
3
4
1
1
1
4
18
2
2
1
1
2
1
5.44
5.37
5.18
4.96
4.58
4.35
4.33
4.11
4.01
4.00
4.00
3.87
3.79
3.75
3.69
3.52
3.51
3.42
3.37
3.37
3.29
3.19
3.15
3.11
3.08
2.99
2.93
2.90
2.89
2.86
2.84
2.75
2.75
2.67
2.65
2.63
2.62
2.61
2.55
2.55
2.47
2.37
2.27
2.17
2.08
1.97
1.91
EAEVVDTSLADPQALcTDK
QLIQWPVEEIETLR
IVVGGDRDNnGmAFLYQSPDFVTWTR
NLIDHSIIESFGAGGK
DNNGMAFLYQSPDFVTWTR
IVVGGDRDnNGMAFLYQSPDFVTWTR
VLWAWVPEKDSEEDDIEK
ENFLPQNGLSLSGSTLDLR
RVLWAWVPEKDSEEDDIEK
ENFLPqnGLSLSGSTLDLR
DNNGmAFLYQSPDFVTWTR
GVFGPFGVLAmASK
nLIDHSIIESFGAGGK
GVFGPFGVLAMASK
DNnGmAFLYQSPDFVTWTR
LEDLKEAEVVDTSLADPQALcTDK
GWAGLqTFPR
GWAGLQTFPR
RVLWAWVPEK
DPSTAWLGPDGLWR
DNnGMAFLYQSPDFVTWTR
QLIqWPVEEIETLR
SGKQLIQWPVEEIETLR
ENFLPQnGLSLSGSTLDLR
IVVGGDRDNNGMAFLYQSPDFVTWTR
DLKEQTAIFFR
TSFGAFVDIDPR
SMVDGILLPSQK
STHNPIITPPEGVK
ISQMSAWSMK
EWVKSTHNPIITPPEGVK
DWYTLGTYSPDR
GASSKGVFGPFGVLAMASK
YSVLMcSDLSR
SmVDGILLPSQK
YDYGNFYASK
YSVLmcSDLSR
QLIQWPVEEIETLRENEVK
DWYTLGTYSPDREnFLPqnGLSLSGSTLDLR
SGSVVEIEGVTASQADVTISFK
DWYTLGTYSPDRENFLPqNGLSLSGSTLDLR
VLWAWVPEK
ISQmSAWSMK
QVQDLAWPK
DWYTLGTYSPDRENFLPQNGLSLSGSTLDLR
QLIqWPVEEIETLREnEVK
SMVDGILLPSQKNEQPYR
a
Charge
2
3
3
2
3
3
2
2
3
2
2
2
3
2
2
3
2
2
3
3
2
2
3
3
3
2
3
2
2
2
3
2
3
2
2
2
2
2
3
2
3
2
2
2
3
3
3
d
MH+ [Da] e
2061.96660
1753.95151
2975.38950
1657.85515
2262.02542
2959.39817
2188.04741
2061.06157
2344.14420
2063.03447
2278.02519
1396.72905
1658.84031
1380.73589
2279.00688
2660.29667
1133.57585
1132.59014
1283.72733
1570.76511
2263.00884
1754.94866
2026.10080
2062.04679
2958.41764
1367.73247
1324.65396
1287.69927
1489.80144
1168.55205
2032.08670
1473.66655
1810.94547
1330.61382
1303.69194
1227.53276
1346.60808
2353.24053
3518.68906
2224.14482
3516.71262
1127.62651
1184.54582
1084.58000
3515.70151
2355.22062
2075.05533
The matched sequences are listed according to the predicted amino acid sequences and the amino
acids modified are in lowercase letters.
b
The total number of identified peptide sequences (PSMs = peptide spectrum matches) for the protein
including those redundantly identified.
c and d
All peptide Xcorr values are above the specified score threshold (the minimum score is 1.3 for a
charge state of 2; the minimum score is 1.305 for a charge state of 3).
e
MH+ [Da] (calculated m/z of the peptide with z = 1).
4
(B)
(A)
50
500
CK Fru
100 mM NaCl Fru
15% PEG6000 Fru
40
-1
umol g DW
-1
umol g DW
400
CK Glc
100 mM NaCl Glc
15% PEG6000 Glc
300
30
200
20
100
10
0
0.5
1
2
3
5
Time (days)
(C)
0
7
1
2
3
5
7
3
5
7
3
5
7
Time (days)
(D)
180
120
CK Suc
100 mM NaCl Suc
15% PEG6000 Suc
100
-1
-1
umol g DW
150
umol g DW
0.5
120
CK 1-K
100 mM NaCl 1-K
15% PEG6000 1-K
80
60
90
40
60
0
0.5
1
2
3
5
7
0
0.5
(E)
125
1
2
Time (days)
Time (days)
(F)
100
CK Nys
100 mM NaCl Nys
15% PEG6000 Nys
80
CK 1F-F
100 mM NaCl 1F-F
15% PEG6000 1F-F
-1
umol g DW
-1
umol g DW
100
75
60
40
50
20
25
0
0.5
1
2
3
5
7
0
0.5
Time (days)
1
2
Time (days)
Fig. S1. Sugar contents in the tubers of Jerusalem artichoke exposed to 100 mM NaCl or 15%
PEG6000 conditions during tuber sprouting. Contents of frutose (A), glucose (B), sucrose (C),
1-kestose (D), nystose (E), and 1F-fructofuranosylnystose (F) were analyzed by HPLC. Samples not
subjected to any treatment were used as the control (CK). Tuber samples were collected at 0, 0.5, 1, 2,
3, 5, and 7 days after treatment, and correspond to those shown in Fig. 8. Values are determined as
µmol g-1DW and represented as means ±SE of three biological samples.
5