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Acyl-CoA oxidase 1 is involved in γ-decalactone release from peach
(Prunus persica) fruit
Liping Zhang · Haiyan Li · Ling Gao · Yujie Qi · Wanyi Fu · Xiongwei Li · Xiang Zhou · Qikang
Gao · Zhongshan Gao · Huijuan Jia
Supplemental material
Table S1 Characteristics of five varieties of peach fruit at the fully ripening stage (S3).
Table S2 The peptides of protein extracted from fully ripe peach mesocarp of ‘HJ’ matched with
the amino acid sequence of PpACX1 and PpACX3 in the peach protein database identified by
EASY-nLC 1000 nanoliter HPLC and Orbitrap Elite MS.
Table S3 Primers of PpACXs for real-time quantitative polymerase chain reaction (qPCR).
Figure S1 The appearance of the fruit of ‘HJ’ and ‘YL’ at four ripening stages. Numbers 0 to 3
after cultivar abbreviations refer to four ripening stages (immature stage, early ripening stage,
physiological ripening stage and fully ripening stage), respectively.
Figure S2 Changes in expression of PpACX genes in mesocarp of peach ‘HJ’ and ‘YL’ at different
ripening stages. For each figure, data shown are means ±SE (n=3). In each figure, asterisks
indicate significant differences in relative expression of PpACX1 at each ripening stage compared
with S0 (*P <0.05; **P <0.01; Student’s t-test). Numbers 0 to 3 after cultivar abbreviations refer
to four ripening stages (immature stage, early ripening stage, physiological ripening stage and
fully ripening stage), respectively.
Figure S3 The primary mass spectrum (A) and secondary mass spectrum (B) of the peptide
segments of protein extracted from fully ripe mesocarp of the peach cv. ‘HJ’.
Figure S4 SDS-PAGE of purified recombinant His-tagged PpACX1. Lane M, protein marker;
lanes 1 and 2, SDS-PAGE of the purified recombinant PpACX1 (15 and 7.5 µg, respectively);
lanes 3 and 4, BSA standard solutions (5.00 and 2.50 µg, respectively).
Figure S5 The primary mass spectrum (A) and secondary mass spectrum (B) of the peptide
segments of peach recombinant PpACX1.
Table S1 Characteristics of five varieties of peach fruit at the fully ripening stage (S3).
Varieties
Fruit type
Flesh
Biological status of
Flesh
Aromatic
Flesh
color
accession
firmness
strength
texturea
(N)
HJ
Peach
White
Improved cultivar
7.0-10.0
Strong
Soft melting
YL
Peach
White
Landrace
＜7.0
Strong
Soft melting
TG
Peach
White
Landrace
13.0-16.0
Intermediate
Hard melting
ZH
Peach
White
Landrace
≥16.0
Slight
Hard melting
ZN16
Nectarine
White
Improved cultivar
≥16.0
Slight
Hard melting
Variety abbreviations: HJ, Hu Jing Mi Lu; YL, Feng Hua Yu Lu; TG, Tai Gu Rou Tao; ZH, Zhong Hua Shou Tao;
ZN16, Zhong Nectarine 16. a According to Wang et al. (2005).
Table S2 The peptides of protein extracted from fully ripe peach mesocarp of ‘HJ’ matched with
the amino acid sequence of PpACX1and PpACX3 in the peach protein database identified by
EASY-nLC 1000 nanoliter HPLC and Orbitrap Elite MS.
Accession Description
1
1
3
Score
44.59
Coverage
17.32
# Proteins
2
# Unique Peptides
9
# Peptides
9
# PSMs
18
Area
6.212E6
# AAs
664
MW [kDa] calc. pI
74.3
8.21
A2
High
Sequence
TDEFVLNSPTLTSSK
# PSMs
4
# Proteins
1
# Protein Groups
1
Protein Group Accessions
1
Modifications
ΔCn
0.0000
Area
5.329E6
XCorr
3.50
Charge
2
MH+ [Da]
1638.81267
ΔM [ppm] RT [min]
-5.80
35.23
High
NLDDHLPLPGITVGDIGmK 3
1
1
1
M18(Oxidation)
0.0000
2.350E5
3.09
3
2021.02567
High
FGNGAYnSmDnGVLR
2
1
1
1
N7(Deamidated); M9(Oxidation);
0.0000
2.880E5
N11(Deamidated)
2.75
2
High
nLDDHLPLPGITVGDIGMK 1
1
1
1
N1(Deamidated)
High
QASLANDQLR
1
2
1
High
QTIVADASTALSR
1
1
High
AATHAVAFAR
2
High
FGnGAYnSmDnGVLR
High
High
-5.76
43.82
1632.68901
-4.24
28.78
0.0000
0.000E0
2.74
3
2006.03409
3.83
47.30
1
0.0000
4.540E6
2.56
2
1115.57634
-3.65
16.11
1
1
0.0000
3.552E6
2.55
2
1332.70647
-4.02
29.96
1
1
1
0.0000
3.129E6
2.55
3
1014.54398
-3.95
11.96
1
1
1
1
4.35
28.69
YVQSNVPR
1
1
1
1
0.0000
3.280E6
2.31
2
962.50170
-3.88
11.88
SLTTSATADAIEEcR
1
1
1
1
C14(Carbamidomethyl)
0.0000
3.118E5
2.20
2
1624.74358
-2.95
27.41
High
LSEEEASWFR
1
1
1
1
0.0000
8.769E6
2.19
2
1253.57085
-7.10
34.81
3
12.37
4.17
1
2
2
7
1.158E6
672
75.1
8.00
A2
High
Sequence
VFVSPDYnQSMEQQR
# PSMs
4
# Proteins
1
# Protein Groups
1
Protein Group Accessions
3
Modifications
N8(Deamidated)
ΔCn
0.0000
Area
0.000E0
XCorr
3.56
Charge
2
MH+ [Da]
1828.82744
ΔM [ppm] RT [min]
5.65
28.11
High
VFVSPDYNQSMEQQR
2
1
1
3
0.0000
5.461E5
3.25
2
1827.81987
-7.23
28.32
High
FGYLSTDNVAAVR
1
1
1
3
0.0000
1.769E6
2.75
2
1412.71147
-3.85
33.79
N3(Deamidated); N7(Deamidated);
0.0000
0.000E0
M9(Oxidation);
2.46
N11(Deamidated)
2
1633.68706
Note: In the column of accession, the numbers 1 and 3 refer to ACX1 and ACX3, respectively.
Table S3 Primers of PpACXs for real-time quantitative polymerase chain reaction (qPCR).
Gene
Product
Primer
Forward primer (5’ to 3’)
Reverse primer (5’ to 3’)
PpACX1
y2510-2
TGAATCAGTTGTGCCCGATG
CCTTTTGTTTCCCATATCCTGG
203
PpACX2
y2282-1
GGAACTGATAGACGCTTTTGAT
ACCAATCCCTTGTTATGCCA
169
PpACX3
Y2439-1
CTAGTGCTGAACTGAAGACCACG
125
PpACX4
Y5916-1
ACTCTGAGGGCAAGGGAAACG
ATTAAACAAAGTTGTCCCGAGAGC
242
PpACX5
Y3524-1
CTTGGGTGCTATGATGGGAATG
AGGAAAGACAGAAGGCTAGTGGG
169
TEF2
TEF2
GGTGTGACGATGAAGAGTGATG
GGTGTGACGATGAAGAGTGATG
129
name
CTGATAATGTTGCTGCTGTGAG
G
Figure S1 The appearance of the fruit of ‘HJ’ and ‘YL’ at four ripening stages.
Figure S2 Changes in expression of PpACX genes (A, B) in mesocarp of peach ‘HJ’ and ‘YL’ at
different ripening stages.
(bp)
Figure S3 The primary mass spectrum (A) and secondary mass spectrum (B) of the peptide
segments of protein extracted from fully ripe mesocarp of the peach ‘HJ’.
A
B
Figure S4 SDS-PAGE of purified recombinant His-tagged PpACX1.
Figure S5 The primary mass spectrum (A) and secondary mass spectrum (B) of the peptide
segments of peach recombinant PpACX1.
A
B