peonidin 3-(coumaroyl)-glucoside. 609. 301,463. 8.2. MR malvidin 3-rutinoside. 639. 331, 493. 8.8. DCRG delphinidin-3-(coumaroyl) rutinoside-5-glucoside.
Supplementary data
Transcription factors, sucrose and sucrose metabolic genes interact to regulate potato phenylpropanoid metabolism
Raja S Payyavula, Rajesh Singh, and Duroy A Navarre*
A
Supplementary Fig. S1.
Payyavula et al., 2013
B
Supplementary Fig. S1 continued.
2
Payyavula et al., 2013
3
C
D
Supplementary Fig. S1 continued. Protein sequence alignment of representative (A) MYBs, (B) bHLH, (C) WD40 and (D) AOMT from potato and from other species. Accession numbers are listed in Fig. 2 or Supplementary Table 3.
Payyavula et al., 2013
a
b
2
White Yellow D.yellow Red Purple
60 a b
40
c
1
d
20
µmol TE g-1
nmoles CA min-1 mg -1 protein
3
4
e c d d
0
0 PAL activity
FRAP
Supplementary Fig. S2. PAL enzyme activity and FRAP antioxidant activity in the five genotypes. The data represents the means ± SE of three biological replicates. Values with same letter are not significantly different (p< 0.05).
Payyavula et al., 2013
5
A
B
Supplementary Fig. S3. (A) StAN1 protein sequence of the five potato genotypes used in this study. Differences are shaded in black. Coding regions were sequenced and translated. (B) StAN1 promoter region from four genotypes used in this study. 1500 bp were sequenced and a 700 bp region containing the SURE and jasmonate motifs starting 500 bp upstream of the start codon were aligned as shown. Methyl jasmonate responsive motifs are highlighted in red. Sugar responsive elements (SURE) are highlighted in green.
Payyavula et al., 2013
Relative expression
0.08
a
a
ab
2.4
a
ab
0.06
cd
ab
0.04
bc
1.6
b
bc bc
6
Wiseman Fairbanks Palmer Juneau Texas Florida
0.8
c
0.02 d cc
0
0 bHLH1
WD40
AN1
Supplementary Fig. S4. Expression of selected transcription factors in purple potatoes grown in in Alaska (Wiseman, Fairbanks, Palmer and Juneau), Texas and Florida. AN1 expression uses the y-axis on the right. The data represents the means ± SE of three biological replicates.
Payyavula et al., 2013
72h
48h AB
7
A
AB
A
Supplementary Fig. S5. Tobacco leaves 48h and 72h after infiltrating with a binary construct harboring AN1 (A) alone or AN1 + bHLH1 (AB).
Payyavula et al., 2013
8
mAU 300
A
AN1
1.0
287.1
E
m/z 595
M/Z 595
449 0 300
AN1
B
m/z 611
0.0 1.0
F
303.1
M/Z 611
465.1 0 AOMT3
300
C
0 300
m/z 625
AOMT3+AN1 m/z 611
D
0.0 1.0
0.0 1.0
449 316.8
G
M/Z 625
478.8
273.8 330.8
H
M/Z 639
m/z 639 286.7
0
492.7
0.0 1
2
3
4
5
6
7
8
9 min
200
300
400
500
600 m/z
Supplementary Fig. S6. MS extracted ion data of the most abundant anthocyanins formed in (A) N. tabacum (Samsun) infiltrated with AN1; (B) N. benthamiana leaves infiltrated with AN1, (C) AOMT3 and (D) AN1+AOMT3. MS2 data for the indicated peaks is shown in panels E-H.
nmoles CA min-1 mg -1 protein
Payyavula et al., 2013
9
6 5 4 3
E B W A
2 1 0 PAL activity
Supplementary Fig. S7. PAL activity in tobacco leaves infiltrated with empty vector (E), bHLH1 (B), WD40 (W) or AN1 (A). The data represents the mean ± SE of three biological replicates.
Payyavula et al., 2013
10
Phn
Phenolics
1
Anthocy 0.95
Anth
1
Asc
1
Phe
1
3CGA
1
C4CGA
1
5CGA
1
4CGA
-0.1
1
Rut
1
Kmp
-0.12
1
BDCS
0.21
1
BCSD
PALe
1
FRAP
1
CHS
1
DFR
1
UFGT
1
AOM12
1
AOM3
0.03
1
PAL
1
HQT
1
HCT
1
C3H
1
AN1
1
AN2
1
M73A
1
M73B
0.47
1
M75B
1
M75C
Ascorbate -0.93 -0.87
Phe -0.65 -0.63 0.37
3CGA -0.07 -0.24 0.09 0.33
C4CGA -0.16 -0.33 0.13 0.44 0.99
5CGA 0.97 0.89 -0.93 -0.67 -0.19 -0.25 4CGA 0.01 -0.18 0.01 0.3
1
0.98
Rut 0.64 0.81 -0.52 -0.37 -0.09 -0.19 0.47 -0.07 Kmp 0.65 0.48 -0.71 -0.5 -0.28 -0.28 BDCS 0.49 0.38 -0.15 -0.81 0.25
0.14
0.81
-0.2
0.43
0.27 0.28
BDCSD -0.19 -0.36 0.3 0.17 0.94 0.92 -0.28 0.92 -0.2 -0.32 0.43 1 Glu Glucose 0.87 0.89 -0.97 -0.27 -0.19 -0.23 0.85 -0.11 0.64 0.55 0 -0.43 1 Suc Sucrose 0.92 0.89 -0.89 -0.69 -0.42 -0.47 PALe 0.97 0.88 -0.93 -0.66 -0.16 -0.22 FRAP
1
0.97 -0.92 -0.67 -0.12
-0.2
0.97 -0.33 0.46 1
0.81 0.33 -0.49 0.83
-0.07 0.45 0.82
0.97 -0.04 0.67 0.63
1
0.43 -0.26 0.84 0.96
0.49 -0.23 0.87 0.92 0.96
CHS 0.97 0.93 -0.93 -0.67 -0.28 -0.36
0.99
-0.21 0.54 0.77
0.36
DFR 0.8
-0.32
0.92
-0.21
0.28 -0.36 0.69 0.91 0.92 0.78 0.89
UFGT 0.97 0.94 -0.93 -0.66 -0.28 -0.35
0.99
-0.2
-0.02
0.66 -0.82 -0.59 -0.3
AOMT1-2 -0.2 -0.45 0.22 -0.15 0.13
0.17
AOMT3 0.92 0.85 -0.91 -0.65 -0.31 -0.36
0.1
0.98
0.55 0.76
-0.4 0.88 0.98 0.98 0.96
0.37 -0.39 0.88 0.98 0.98 0.97 0.99 0.88
0.14 -0.83 0.48
0.23
0.3
0.99 -0.22 0.37 0.87
0.34
-0.4 0.83 0.99 0.98 0.92 0.98 0.96 0.98
-0.4 -0.1
0
0.39 -0.38 0.87 0.99 0.99 0.97
PAL 0.97 0.93 -0.93 -0.68 -0.28 -0.35
0.99
HQT 0.07 -0.1 -0.08 0.32 0.98
0.97
-0.05 0.99 0.02 -0.19 0.23 0.87
HCT 0.44 0.17 -0.5 -0.32 0.06
0.08
0.6
0.14 -0.41 0.91 0.25 0.05 0.28 0.53 0.62
C3H 0.65 0.42 -0.56 -0.69 0.02
-0.02
0.76
0.09 -0.11 0.88
AN1 0.9
0.81 -0.88 -0.67 -0.31 -0.36
AN2 -0.24 -0.33 0.55 -0.49 0.09
0.54 0.77
0.98 -0.23 0.31
0.9
-0
1
0.89
-0.3 -0.02 0.02 -0.2 -0.2
1
-0.11 0.98
-0.14
0.04
-0.17 -0.1
0.5 0.82 0.49
0.73
0.64
0.62 0.08 0.34 0.69 0.78 0.62 0.67 0.86 0.67
0.61
0.76 0.69 0.06 0.89
0.36 -0.39 0.79 0.98 0.98 0.89 0.97 0.98 0.96
0.11
0.4
1
0.51 0.1
0.97 -0.2 0.69 0.81
-0.22 0.05 -0.31 -0.09 0.69 0.42 -0.7 -0.2 -0.22 -0.24 -0.3 -0.2 -0.28
0.56
-0.24 -0.3
MYB73A -0.56 -0.49 0.39 0.31 -0.67 -0.56
-0.4
0.25
-0.24 -0.4 -0.7
MYB73B 0.04 -0.1
0.25 -0.43 -0.56 0.69
0.8
0.36 0.22 -0.5 0.69 0.67 0.43
0
MYB75B -0.02 0.06 0.03
0.04
-0.2
-0.24 -0.1 0.28 -0.15
-0.48 -0.46 -0.44 0.4
0.68
0.64
-0.69 -0.59 0.04 -0.68 -0.61 -0.3 -0.2 -0.41 -0.55 -0.3 -0.1 -0.35
-0.24 0.65
0.5
-0.66
0.21 -0.28 -0.2 0.33 0.26 0.03 0.21 0.56 0
0.2
-0.1 0.07 0.28 -0.18 -0
-0.27 -0.21 -0.08 0.47
0.53 0.07 -0.4 -0.23 -0.02 -0.2 -0.6 -0.22 -0.63 -0.37 -0.2 0.71 -0.6 -0.51 -0.43 -0.28
-0.66 -0.95
MYB75C -0.41 -0.4 0.67 -0.42 -0.27
-0.3
-0.35 -0.32 -0.35 -0.16 0.42 0.06 -0.7 -0.3 -0.37 -0.39 -0.4 -0.2 -0.37
0.45
-0.32 -0.4 -0.4
0.54 -0.47
1
M12B
MYB12B -0.73 -0.81 0.65 0.75 0.73
0.79
-0.78 0.67 -0.51 -0.61 -0.26 0.73 -0.7 -0.9 -0.76 -0.76 -0.9 -0.7 -0.84
0.2
-0.82 -0.8 0.63 -0.2 -0.45 -0.81 0.12
-0.03 -0.36 0.48
0
1
bHL1
0.98
-0.2 0.48 0.66 0.96 -0.28
-0.35
0.19 -0.22 -0.36 -0.85
1
bHL2
0.55 0.42 -0.6 0.65 0.45 0.57 -0.28
0.63
0.71 -0.83 -0.1 -0.5
0.4
1
W40
bHLH1 0.97 0.94 -0.93 -0.67 -0.28 -0.36
0.99
bHLH2 0.2
0.42 -0.56 -0.38 0.79 -0.35 -0.67 0.34 0.53 0.41 0.19 0.42 0.71 0.41
0.13 -0.4 -0.12 -0.62 -0.55
WD40 0.94 0.86 -0.91 -0.68 -0.25
-0.3
1
-0.21 0.56 0.75
-0.16 0.4
0.86
0.37
0.4
-0.4 0.89 0.99 0.98 0.97
1
0.88
1
-0.33 0.82 0.98 0.99 0.94 0.98 0.95 0.98
-0.16 0.4 0.04
1
1
-0.1 0.07 -0.26
0.99 -0.1 0.65 0.79 0.99
0.91
-0.2
0.28
-0.32 0.33 -0.33 -0.31 -0.8 0.98 0.49
magnitude of change 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.89 >0.99 greater than zero less than zero
Supplementary Fig. S8. Correlation analysis of transcript and metabolite expression in tubers from five potato genotypes. Pearson correlation coefficients are shown.
1
Payyavula et al., 2013
11
Supplementary Table S1. Retention times and MS data of select compounds present in potato extracts separated by HPLC. RT
Abbreviation Compound
m/z
MS/MS (m/z )
1.8
Asc
ascorbic acid
175 (M-H)
115, 87, 71
2.5
Tyr
tyrosine
180 (M-H)
163, 119, 93
4.0
Phe
phenylalanine
164 (M-H)
147, 148, 72
5.9
Trp
tryptophan
203 (M-H)
159, 116, 142, 186
6.5
3CGA
3-o -chlorogenic acid
353 (M-H)
191, 179, 135
7.0
CP
caffeoyl putrescine
251 (M+H)
234, 163, 89, 115
7.6
C4CGA
cis-4-chlorogenic acid
353 (M-H)
173, 179, 135, 191
8.3
5CGA
5-o -chlorogenic acid
353 (M-H)
191,179, 161,135
8.4
BDCS
bis-dihydrocaffeoyl spermine
531 (M+H)
293, 222, 513, 165, 367
8.5
4CGA
4-o -chlorogenic acid
353 (M-H)
173, 179, 191, 135
9.1
C5CGA
cis-5-chlorogenic acid
353 (M-H)
191, 179, 173
9.6
BDCSD
bis-dihydrocaffeoyl spermidine
474 (M+H)
222, 457, 165, 310, 236
10.4
FQA1
feruloyl quinic acid1
367 (M-H)
161, 135, 335
11.5
FQA2
feruloyl quinic acid2
367 (M-H)
179, 135, 191, 161, 143
12.0
TDCS
tris-dihydrocaffeoyl spermine
695 (M+H)
293, 222, 531, 474, 457
12.3
QRG
quercetin rutinoside-glucoside
771 (M-H)
591,300, 301, 609
12.6
QDG
quercetin diglucoside
625 (M-H)
300, 301, 445 607, 463
12.8
Myr
myricetin rutinoside
625 (M-H)
317, 316, 287, 607,461, 271
13.4
KRG
kaempferol rutinoside-glucoside
755 (M-H)
575, 285, 593
13.5
TDCSD
tris-dihydrocaffeoyl spermidine
638 (M+H)
456, 474, 293, 222
14.0
Rut
quercetin 3-rutinoside
609 (M-H)
301, 609, 271, 343
14.6
Kmp
kaempferol 3-rutinoside
593 (M-H)
285, 593, 431
14.8
RR
rhamnetin rutinoside
623 (M-H)
315, 300, 316, 271
15.4
Glyc
solanine
852 (M+H)
706, 398, 560
15.4
Glyc
chaconine
868 (M+H)
398, 706, 722, 560, 380
Payyavula et al., 2013
12
Supplementary Table S2. Retention times and MS data of anthocyanins in potato phenolic extracts separated by HPLC. RT
Abbreviation Com pound
[M] + m /z
MS/MS (m /z)
3.5
PtRG
petunidin 3-rutinoside-5-glucoside
787
625,317,479
3.8
PlRG
pelargonidin 3-rutinoside-5-glucoside
741
579,271,433
3.9
DR
delphinidin 3-rutinoside
611
303, 465, 449
4.6
DRG
delphinidin 3-rutinoside-glucoside
773
303,611,465
5
CR
cynanidin 3-rutinoside
595
287,449
5.2
MRG
malvidin 3-rutinoside-5-glucoside
801
639,331,493
5.4
PlG
pelargonidin 3-glucoside
433
271
6.1
PlDGG
pelargonidin 3-diglucoside-5-glucoside
757
271,595
6.2
PeR
pelargonidin 3-rutinoside
579
271,433
6.3
PtR
petunidin 3-rutinoside
625
317,479
6.5
PlR
pelargonidin 3-rutinoside
579
271,433
7.4
PoCG
peonidin 3-(coumaroyl)-glucoside
609
301,463
8.2
MR
malvidin 3-rutinoside
639
331, 493
8.8
DCRG
delphinidin-3-(coumaroyl) rutinoside-5-glucoside
919
303,757,465
8.9
PlCRG
pelargonidin 3-(coumaryol) rutinoside-5-glucoside
887
725,271,433
9.06 PtCfRG
petunidin 3-(caffeoyl) rutinoside-5-glucoside
949
787,317,479
9.3
PlCRG
pelargonidin 3-(coumaroyl) rutinoside-5-glucoside
887
725,271,433
9.7
PeCRG
peonidin 3-(coumaroyl) rutinoside-5-glucoside
917
301,755,463
9.8
PlCDG
pelargonidin 3-(coumaroyl) diglucoside-5-glucoside
903
741,271,433
10.1 CCRG
cyanidin 3-(coumaroyl) rutinoside-5-glucoside
903
741,287,449
10.3 PtCRG
petunidin 3-(coumaroyl) rutinoside-5-glucoside
933
317,771,479
11
pelargonidin 3-(coumaroyl) rutinoside-5-glucoside
887
725,271,433
11.1 PtFRG
petunidin 3-(feruloyl) rutinoside-5-glucoside
963
317,801,479
11.4 PeCRG
peonidin 3-(coumaroyl) rutinoside-5-glucoside
917
755,301,463
11.8 MCRG
malvidin 3-(coumaroyl) rutinoside-5-glucoside
947
785,331,493
12
pelargonidin 3-(feruloyl) rutinoside-5-glucoside
917
755,271,433
12.3 PtCR
petunidin 3-(coumaroyl)-rutinoside
771
317,479,245
12.5 MFRG
malvidin 3-(feruloyl) rutinoside-5-glucoside
977
331,815,493
13.3 PlRG
pelargonidin rutinoside-5-glucoside
725
271,433
13.7 PeRG
peonidin rutinoside-5-glucoside
755
301,463
PlCRG
PlFRG
Payyavula et al., 2013 Supplementary Table S3. List of primers used in this study. Species Primer
Potato
Vector
StMYB73A-F StMYB73A-R StMYB73B-F StMYB73B-R StMYB12A-F StMYB12A-R StMYB12B-F StMYB12B-R StAN1-F StAN1-R StAN2-F StAN2-R StMYB75A-F StMYB75A-R StMYB75B-F StMYB75B-R StMYB75C-F StMYB75C-R StMTF1-F StMTF1-R StMTF2-F StMTF2-R StMYB10-F StMYB10-R StWD40-F StWD40-R StbHLH1-F2 StbHLH1-R2 StbHLH2-F2 StbHLH2-R2 StAOMT-1/2F StAOMT-1/2R StAOMT-3F StAOMT-3R StAN1-F-FL StAN1-R-FL StbHLH1-F-FL StbHLH1-R-FL StWD40-F-FL StWD40-R-FL 35S-F pORE-F pORE-R
Primer sequence GCCAGACAAGGTATTTGTGCCGTT TGCGATTAGCAGCTCTATCCCTGA GCCGGTGGTTGATAAACCGATTCC GCATTCCTTATTGCATCRGTCTGC TGTTTATGGGACGGAGAAGGAG GCCAAGTTACCAAATCACTCTGCATGG AGGAGATTATGCCAGACGTGGTGA CATGGTGGCGTTGGCCATACTAAT AAGTATGGGCAAGCCAATGCCA GCCCACCATTGCATATCGTTGTTGTC AGACCTCAACCTCGGAACTTCTCA GTCCACCATTGAACTCCATCGTCT CACTCCTCAAGTTGTGTTTGTCCC TGAAGCTATGTTGCTCGGGCTCTT TGCACCAAAGCCCATAAGGATTTC GCAGCTTTCTCCCATAAGAATTTCA TGGTCCCTCATTGCCGGAAGAATA GCGGCTCATTTCCTTGTCCTTTGA AGCAAACTCCAGATGCATTGATGG CCTTGTTGCATGGAGTTACCTTCACC TGCTGAAAGCAAAGGTGGAATGGG TAATAGCCTCAGTTTCAGCCTCAGGG GCATCAAGATTGCCAGGAAGAACAG AGTGGATGGTAGATCATTGTGTTGT ATTCAAGTCAAGAATCGCATCTCCG GAACTCTTCGATAAAGCTYCCGAC TCTCTTGGAYGGTGTAGTGGAACT AACAGCGGATGACGGAGTATTGCT AGGCSAGATCAGCAAGACGATCAA AGCTCTCGCTCCAAAGACAACTCA CCTGTTCTTGACAAACTCCTGGAAG TCAYCAGCTKCATTAGTCTCTCGT GGTGAAAGTTGGAGGAGTACTTGGCT TGCCTTTAATCCTGGTGGCATAGG AATGCGGCCGCATGAGTACTCCTATGATGTGTAC CCGGGTACCTTAATTAAGTAGATTCCATATATC AATGCGGCCGCATGGAGATTATACAGCCTAATAGC CCGGGTACCTTAATTAGCTCTAGGGATTATCTG AATGCGGCCGCATGGARAATTCAAGTCAAGAATCG CCGGGTACCTTATACTTTAAGCAGCTGCAACTTG CAAGACCCTTCCTCTATAT ACTGAAGGCGGGAAACGACAATCT AGCGGATAACAATTTCACACAGG
Used for
Accession no or Potato Genome Contig PGSC0003DMS000002386 PGSC0003DMS000003180 PGSC0003DMS000001330 PGSC0003DMS000000077 PGSC0003DMS000000121 PGSC0003DMS000000777 PGSC0003DMS000000511 PGSC0003DMT400031607
QPCR
PGSC0003DMT400085829 EU310399 PGSC0003DMS000000013 PGSC0003DMT400060168 PGSC0003DMT400001513 PGSC0003DMT400033569 PGSC0003DMT400032139 PGSC0003DMT400006134 and PGSC0003DMT400006136 PGSC0003DMT400016512
Cloning
13
Payyavula et al., 2013 Supplementary Table S3 continued. Species Primer
Primer sequence
NtINV1-F TTAGAGACCCAACGACTGCTTGGA NtINV1-R ACCCGTTTGCCTCATCGGTTGATA NtSUSY-F TTCACCTCAYGGATATTTCGCCCA NtSUSY-R AGTCCTTGCTCCTTTATGCGCT NtCHS-F ACTTGGYAAAGAGGCWGCCCAA NtCHS-R CGGCAAAGCAACCTTGTTGGTA NtPAL1/4-F TTCGCCTACGCTGATGATCCTT NtPAL1/4-R TGCAGCTCTTGCACTCTCAACT NtPAL2/3-F ACCCACTGATGCAGAAACTAAGGC NtPAL2/3-R GCAATTGCAGGGTTCCCAYTTT NtDFR-F ATTCATCTGCGCRTCCCATCAT NtDFR-R AAATACACCACRGGCAAGTCCT Tobacco NtF3H-F AGGCYTTAACCAAGGCATGTGT NtF3H-R CGGGCTGAACAGTRATCCAAGT NtBHLH1-F ATGKGCGCAAACGAGGTTGATAGC NtBHLH1-R TRGCTGAGGTTGTTGTTGCTCA NtINV1-F TTAGAGACCCAACGACTGCTTGGA NtINV1-R ACCCGTTTGCCTCATCGGTTGATA NtINV2-F TATACCGGGTCAACCAACGAGTCA NtINV2-R CATTTGCCTTGTGGTGTGGTCCAT NtActin-F AAGCTGTGTTGTCCCTATACGCCA NtActin-R ATCGCGGACAATTTCCCGTTCA NtELF-F TGAACCATCCAKGACAGATTGG NtELF-R TGGGCTCCTTCTCAATCTCCTT
Used for
Accession no AJ305044 AB055497 AF311783 M84466/EU883669 D17467/X78269 AB289448
QPCR
AB289450 HQ589208/HQ589209 AJ305044 HM022269 AB158612 D63396
14
Payyavula et al., 2013
15
Supplementary Table 4. Protein similarity matrix of transcription factors from different species. (A) MYB (B) bHLH and (W) WD40. Accession numbers are listed in Fig. 2.
Payyavula et al., 2013
16
Supplementary Table S5. List of regulatory elements in promoters of different genes from S. tuberosum group Phureja. The numbers in each column represent the position of the element in the promoter sequence relative to the ATG start codon. Promoter elements (sequence) Genes
MYBCORE (CNGTTR)
MYBPLANT (MACCWAMC)
MYCCONSENSUS (CANNTG)
SURE (AATAGAAAA)
AN1
-1380, -1279, -786
none
-1349, -13
-604, -561, -521
bHLH1
-737, -703
-949
-1466, -1154, -960, -510, -409
none
WD40
-1422, -1132, -1096, -908, -678, -437, -133 -63
-1150, -1123, -956, -711
-214
PAL1
-825, -271, -268
-1404, -337
-1385, -1346, -189
none
HQT
-1021, -569
none
-1299
-1082
C3H
-1368, -1329, -1217, -1079, -1067, -576
none
-1271, -1238, -1211, -1067, -1040, -614, -576, -526
none
HCT
-92
none
-872, -656, -597
-178
CHS
-366
none
-1336, -1171, -200, -178
none
F3H
-1082
none
-477, -335, -137
none
DFR
-1395, -1207, -1106, -1039, -874
-189
-1236, -1135, -884, -813, -561, -403, -331, -160, -46 none
ANS
-1330, -1203, -747, -628, -625
-386, -171
-747, -617, -521, -344, -227
none
SUSY1
-1396, -1365, -573, -541, -445, -51
-1299, -1086, -645
-1257, -1067, -998, -846, -730, -685, -7
-1279, -666
SUSY4
-1012, -271, -240, -202
-848
-1289, -1132, -650, -444, -240, -214, -202
none
INV1
none
-1442, -203
-1396, -1373, -1339, -1142, -1130, -1079, -1049
none
INV2
-717, -548, -83
-539, -476, -259, -198
-980, -622, -124, -83
none