Chapter 7
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Effects of Germplasm Origin and Fruit Character on Volatile Composition of Peaches and Nectarines Yiju Wang,a,b Feng Chen,c Jinbao Fang,d Chunxiang Yang,a Jianbo Zhao,e Quan Jiang,e and Shaohua Lif,* aInstitute
of Botany, The Chinese Academy of Sciences, Beijing 100093, P.R. China bGraduate School of Chinese Academy of Sciences, Beijing 100049, P.R. China cDepartment of Food Science and Human Nutrition, Clemson University, Clemson, SC 29634, USA dZhengzhou Fruit Institute, The Chinese Academy of Agriculture Sciences, Zhengzhou 450009, P.R. China eInstitute of Forestry and Pomology, Beijing Academy of Agriculture and Forestry Science, Beijing 100093, P.R. China fWuhan Botanical Garden, The Chinese Academy of Sciences, Wuhan 430074, P.R. China *Corresponding author. Telephone: +86-27-87510599. Fax: +86-27-87510251. E-mail:
[email protected].
Ninety-five volatile chemicals in peaches and nectarines with different fruit characters representing different germplasm origins were investigated using HS-SPME-GC-MS. The result showed that the composition and content of volatiles depended on germplasm origins and fruit characters. Chinese bred cultivars obtained from Chinese local and foreign cultivars and Japanese cultivars had significantly higher contents of total volatiles and esters than Chinese local cultivars. White-flesh flat peaches had the highest contents of total volatiles and esters, which were significantly higher than those of other groups with different fruit characters. Terpenoids in white-flesh flat peaches was significantly higher than that in white-flesh nectarines. Ten cultivars including ‘Yutian’, ‘Ruipan 3’, ‘Yangzhou 124 © 2010 American Chemical Society Qian and Rimando; Flavor and Health Benefits of Small Fruits ACS Symposium Series; American Chemical Society: Washington, DC, 2010.
Pantao’, ‘Beijing 5’, ‘Fenglu’, ‘Kanto 5’, ‘Okitsu’, ‘Charme’, ‘Babygold 5’ and ‘Babygold 6’ with high contents of lactones, terpenoids and total volatiles are considered the desirable cultivars that can be used for further breeding.
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Introduction It is well known that peaches and nectarines are native to China where peaches have been cultivated for at least 3000 years, and nectarines for over 2000 years. Peach tree was introduced into Europe at the beginning of the Roman era and then into the United States during the 19th century. With a long history of cultivation and extensive geographical distribution, there are a rich germplasm resources of peaches and nectarines in China and all over the world. However, regardless of the significant development of novel cultivars and increase of fruit production, many peaches and nectarines are often criticized about their undesirable qualities, which were affected by size, external color, texture (e.g., firmness), and contents of sugars, acids, as well as aromas. As one of the essential indices of fruit quality, aroma-impact flavors of peaches and nectarines have been intensively studied, resulting in the identification of more than 110 volatile compounds (1–11), which include C6 compounds, alcohols, aldehydes, esters, terpenoids, ketones and lactones. Among the lactones, particularly γ- and δ-decalactones, have been reported to be the major contributors to the peach aroma, while other volatiles such as C6 aldehydes, alcohols and terpenoids, also made a certain degree of contribution (12–15). Moreover, intensive investigations have focused on the aroma evolution of peach and nectarine during ripening (3, 7, 8, 12, 15, 16) and cold storage (9, 17). It is well known that the composition and content of volatile aromas have significant changes during the maturation. For example, in immature fruits, C6 compounds are the initial major aroma contributors, but their levels decrease drastically while those of lactones, benzaldehyde and linalool increase significantly during maturation (5, 12). Several studies have also investigated the effect of culturing techniques and managements on composition and content of volatiles, which could be changed by orchard management, such as fertilization (18), storage (19), climate or microclimate conditions such as sun light (20), and postharvest treatment (4, 7, 21, 22). Volatile composition is also cultivar dependent. Engel et al. concluded that nectarines contained significantly higher amounts of δ-decalactone than peaches (5, 13). Robertson et al. reported that white-flesh peaches contained more linalool than yellow-flesh cultivars (23). Headspace solid-phase microextraction (HS-SPME) was a sampling technique based on sorption of analytes on a polymeric material that is coated on a silica fiber. It has been widely used for the analyses of volatile and semi-volatile compounds of food and fruits (11, 24–30) since its introduction by Zhang and Pawliszyn in 1993 (31). The HS-SPME technique has the following advantages: low cost, small amount of sample required, no solvent contamination, short time of extraction, simplicity, high selectivity and sensibility (32). The HP-SPME 96 Qian and Rimando; Flavor and Health Benefits of Small Fruits ACS Symposium Series; American Chemical Society: Washington, DC, 2010.
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method involves two steps: the partitioning of the analytes between the coating material and the sample; and the thermal desorption of the analytes into gas chromatograph (28). In this study, HP-SPME combined with gas chromatography-mass spectrometry (GC-MS) was applied to study the characteristic volatiles of peaches and nectarines at the germplasm level. This study mainly focused on comparing the aroma composition and content among the germplasm resources, which include Chinese local cultivars and a number of cultivars from other countries in order to acquire information for future breeding efforts aiming at enhancing fruit quality via effects on aroma.
Materials and Methods Plant Materials A total of 95 peach (Prunus persica L. Batsch) cultivars were used to study the aroma composition and content (Table 1). All the samples were collected from the Germplasm Repository for peach in the Institute of Forestry and Pomology, Beijing Academy of Agriculture and Forestry Science, in 2007. The trees, grafted on a wild P. persica rootstock, were planted in 3 m apart within rows and 5 m apart between rows in the spring of 1996. They were trained to ‘Y’ training systems and pruned by the long pruning method in winter (33). The same orchard managements, such as fertilization and irrigation were applied in the whole orchard. The fruits were picked from three trees of each cultivar when the green color of the fruit skin has almost disappeared. At the same time, the ground color of white-fleshed peaches turned milk white, whilst the ground color of yellow-fleshed peaches turned yellow or orange. All the fruits were picked from the southern or western crown about 1.5-2 m high from the ground of the tree at maturity. The fruit samples were taken to the laboratory immediately after harvest, washed by deionized water and surface-dried with gauze. Then three slices were taken from different orientations of each fruit. Three fruits were used for one composite sample per tree, and considered as one replication resulting in three replications for every cultivar. Samples were ground to a powder in liquid nitrogen and stored at -40°C until analysis.
Isolation and Concentration of Volatiles For headspace sampling, SPME fibers coated with poly-dimethylsiloxanedivinylbenzene (65 µm, PDMS/DVB) (Supelco Co., Bellefonte, PA, USA) were used by optimization of the method carried out in a previous work (24). The fiber was activated according to the manufacturer’s instructions. The SPME method was used for the isolation and concentration of volatiles. For each extraction, 2 g of the pulp powder, 40 µL 3-octanol (0.814 mg L-1, added as an internal standard), 0.6 g NaCl and 0.4 g CaCl2 (added to increase partitioning of volatiles from the 97 Qian and Rimando; Flavor and Health Benefits of Small Fruits ACS Symposium Series; American Chemical Society: Washington, DC, 2010.
liquid into the gas phase and to restrain the enzyme activity, respectively) (34, 35) were placed in a 4 mL capped vial. The vial was placed in a 45°C water bath with a consistent magnetic stirring when the SPME fiber was exposed to the headspace of the sample to adsorb the volatile analytes for 30 min. The fiber was then introduced into the GC injector port for desorption at 220°C for 2 min in the splitless mode. Linear retention index (LRI) were calculated by using n-alkane standards (C5-C40) based on the method of Ceva-Antunes, et al. (36).
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Table 1. Peach and nectarine cultivars are listed according to their origins. The number in parenthesis following the cultivar indicates the accession number. Populations
Characters WF
Cla
Cb
CF
J
b
Taxa Baimangpantao (1)
WP
Baifentao (2), Baiyintao (3), Cuixiangtao (4), Jinhongtao (5), Linbai 4 (6), Qingmaozibaihuatao (7), Xiaobaihua (8)
YP
Huanglu (9), Zaoshenghuangjin (10)
WF
Ruipan 4 (11), Ruipan 5 (12)
WP
Baihuatao (13), Ji 2102 (14), Lvhua 7 (15), Qiuxiangmi (16), Wanshuomi (17), Xinbaihua (18), Yulu (19), Yutian (20), Zaobaimi (21)
YP
Chengyan (22), Jinshiji (23), Wanshiji (24)
WF
Ruipan 3 (25), Yangzhou124 Pantao (26)
WP
Beijing 5 (27), Fenglu (28), Huayu (29), Jingyu (30), Qinxuan 2 (31), Wanhongmi (32), Xiahui 6 (33), Yihongshuimi (34), Zaoyu (35)
YP
Guihuang (36), Qiukui (37), Yanfeng (38)
WN
Ruiguang 2 (39), Ruiguang 7 (40), Ruiguang 27 (41)
YN
81-26-4 (42), 90-8-18 (43), 94-6-19 (44)
WP
Hakuto (45), Nippon Suimitsu (46), Okayama 500 (47), Shin Musume (48), To Hakuho (49)
YP
Ban Ougon (50), Kanto 5 (51), Myojo (52)
YN
Okitsu (53)
WP
Charme (54), Chimarrita (55), Corol (56), Fuzalode (57), J7 (58), NJ250 (59), NJ257 (60), P6 (61) Continued on next page.
98 Qian and Rimando; Flavor and Health Benefits of Small Fruits ACS Symposium Series; American Chemical Society: Washington, DC, 2010.
Table 1. (Continued). Peach and nectarine cultivars are listed according to their origins. The number in parenthesis following the cultivar indicates the accession number. Populations
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AE
Characters
Taxa
YP
Androreda (62), Babygold 5 (63), Babygold 6 (64), Babygold 9 (65), Bulgaria 2(66), Caullinan (67), Dixon (68), Early Crawford (69), Emilia (70), Fortuna (71), Fuzador (72), Glohaven (73), Harbrita (74), Havis (75), Loring (76), Mcneely (77), NJC47 (78), NJC77 (79), NJC237 (80), Norman (81), Redtop (82), Riogrand (83), Shasta (84), Vivian (85)
YN
Flavortop (86), French Nectarine (87), Great Diamond (88), Legrand (89), NJN78 (90), Nectared 4 (91), Nectared 6 (92), P1 (93), Tasiva (94), Vega (95)
a
Cl Chinese local cultivars; Cb pure Chinese original bred cultivars; CF China × foreign cultivars; J Japanese cultivars; AE American and European cultivars b WF= white-flesh flat peaches; WP= white-flesh peaches; YP= yellow-flesh peaches; WN= white-flesh nectarines; YN= yellow-flesh nectarines
GC-MS Conditions The volatile constituents were analyzed by an Agilent (Palo Alto, CA) 5975 mass selective detector coupled to an Agilent 7890 gas chromatograph, equipped with a 30 m × 0.25 mm ×1.0 µm HP-5 MS (5% phenyl-polymethylsiloxane) capillary column. Helium was used as the carrier gas at a linear velocity of 1.0 mL/min. The injector temperature was kept at 220°C and the detector at 280°C. The oven temperature was programmed from 40°C (2 min), increasing at 3°C /min to 150°C (2 min), then increasing at 10 °C /min to 220 °C, and then held for 2 min by optimizing the method used in the previous investigations (2, 25). Mass spectra were recorded in the electron impact (EI) ionization mode at 70 eV. The quadrupole mass detector, ion source and transfer line temperatures were set at 150, 230 and 350°C, respectively. Mass spectra were scanned in the range of m/z 30-350 amu at 1 s intervals. Identification of volatile compounds was achieved by comparing the mass spectra of our collected standards and those of the data system library (NIST 05) and linear retention index.
Statistical Analysis All the cultivars were classified according to two methods: 10 Chinese local (Cl), 14 pure Chinese bred (Cb, cultivars obtained from the cross between two Chinese local cultivars), 20 China × foreign (CF, Chinese bred cultivars containing blood of foreign cultivars), 9 Japanese (J) and 42 American and European cultivars (AE) according to origins; 5 white-flesh flat peaches (WF), 38 white-flesh peaches (WP), 35 yellow-flesh peaches (YP), 3 white-flesh nectarines (WN) and 8 yellow99 Qian and Rimando; Flavor and Health Benefits of Small Fruits ACS Symposium Series; American Chemical Society: Washington, DC, 2010.
flesh nectarines (YN) according to the color of fruit flesh, the fruit shape and with or without hair on the fruit surface (Table 1). Data for each cultivar were averages of three replications. A one-way ANOVA analysis was used to determine significant differences of volatiles contents between groups.
Results and Discussion
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Identification of Volatiles Seventy-five volatile compounds were identified and relatively quantified. Some volatiles were only found in a few cultivars in this study (Table 2). Those compounds included 5 C6 compounds, 8 esters, 13 aldehydes, 8 lactones, 17 terpenoids, 4 ketones, 13 alcohols and 7 other compounds including some carbonyl compounds.
Table 2. Volatiles detected in fruits of all 95 peaches and nectarines C6 compounds
Lactones
LRIb
Compoundsa
Codes
Hexanal
C1
802
2-Hexenal
C2
847
(Z)-3-Hexen-1-ol
C3
851
(E)-2-Hexen-1-ol
C4
862
1-Hexanol
C5
870
γ-Hexalactone
L1
1051
γ-Heptalactone
L2
1151
γ-Octalactone
L3
1255
γ-Nonalactone
L4
1359
6-Amyl-α-pyrone
L5
1454
γ-Decalactone
L6
1464
δ-Decalactone
L7
1490
δ-Dodecalactone
L8
1680
Continued on next page.
100 Qian and Rimando; Flavor and Health Benefits of Small Fruits ACS Symposium Series; American Chemical Society: Washington, DC, 2010.
Table 2. (Continued). Volatiles detected in fruits of all 95 peaches and nectarines
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Aldehydes
Esters
Terpenoids
LRIb
Compoundsa
Codes
Pentanal
A1
697
(E)-2-Pentenal
A2
752
Heptanal
A3
902
Benzaldehyde
A4
955
Octanal
A5
1003
Benzeneacetaldehyde
A6
1039
(E)-2-Octenal
A7
1056
Nonanal
A8
1103
(E)-2-Nonenal
A9
1158
Decanal
A10
1204
(E)-2-Decenal
A11
1238
2,4-Decadienal
A12
1314
Undecanal
A13
1362
Butyl acetate
E1
812
(Z)-3-Hexenyl acetate
E2
1009
Hexyl acetate
E3
1016
2-Hexenyl acetate
E4
1018
Methyl 2-methylpentanoate
E5
1220
3-Hydroxy-2,4,4-trimethylpentyl 2-methylpropanoate
E6
1346
Diisobutyl phthalate
E7
1872
Dibutyl phthalate
E8
1969
D-Limonene
T1
1024
cis-Linalool Oxide
T2
1086
Linalool
T3
1099
Oxoisophorone
T4
1140
(3E)-5-Ethyl-6-methyl-3-hepten-2-one
T5
1145
Epoxylinalol
T6
1166
α-Terpineol
T7
1186
p-Menth-1-en-9-al
T8
1212
α-Citral
T9
1270
Continued on next page.
101 Qian and Rimando; Flavor and Health Benefits of Small Fruits ACS Symposium Series; American Chemical Society: Washington, DC, 2010.
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Table 2. (Continued). Volatiles detected in fruits of all 95 peaches and nectarines
Alcohols
Ketones
Compoundsa
Codes
LRIb
(E)-Theaspirane
T10
1309
α -Damascenone
T11
1380
α, α-Dihydro-á-ionone
T12
1433
4-(2,6,6-Trimethyl-cyclohex-1-enyl)-butan-2-ol
T13
1441
Geranyl acetone
T14
1451
2,6-Ditert-butylquinone
T15
1460
α -Ionone
T16
1481
Nerolidol
T17
1747
1-Penten-3-ol
B1
684
Isoamylol
B2
730
1-Pentanol
B3
768
1-Octen-3-ol
B4
980
2-Ethyl-1-hexanol
B5
1030
5-Methyl-5-nonanol
B6
1121
3-Methyl-3-nonanol
B7
1138
6-Tridecanol
B8
1317
4-Tridecanol
B9
1339
10-Dodecyn-1-ol
B10
1446
6-Pentadecanol
B11
1519
4-Pentadecanol
B12
1549
1-Hexadecanol
B13
1594
1-Penten-3-one
K1
684
Cyclohexanone
K2
895
1-Octen-3-one
K3
978
6-Methyl-5-hepten-2-one
K4
988
Continued on next page.
102 Qian and Rimando; Flavor and Health Benefits of Small Fruits ACS Symposium Series; American Chemical Society: Washington, DC, 2010.
Table 2. (Continued). Volatiles detected in fruits of all 95 peaches and nectarines
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Others
LRIb
Compoundsa
Codes
2-Pentyl-furan
O1
991
Butylated hydroxytoluene
O2
1511
2,4-Di-tert-butylphenol
O3
1514
Hexadecane
O4
1600
Heptadecane
O5
1700
3,5-Di-tert-butyl-4-hydroxybenzaldehyde
O6
1767
Nonadecane
O7
1900
a
Identities confirmed by comparing mass spectra and retention time with those of authentic standards. b Linear retention index calculated using a series of n- alkane.
Table 3. Average contents of major compounds and the groups of volatiles (µgkg-1 FW equivalent of 3-octanol) in different origins of peach and nectarine cultivars Cla
J
AE
128.0±14.4bb 145.6±18.3ab 190.0±16.2a
132.2±7.1b
175.9±9.2ab
C2
31.9±9.1
32.2±5.9
38.9±7.1
30.9±5.7
20.5±2.8
C3
20.3±5.2b
38.7±8.7a
38.6±4.9a
29.4±5.0ab
21.3±2.0b
C4
8.3±2.1ab
10.8±1.8ab
12.3±1.8a
9.4±1.2ab
6.6±0.6b
C5
18.9±5.1ab
30.9±5.8ab
33.3±6.0a
27.1±4.4ab
17.9±1.7b
L1
25.0±9.4
23.6±6.3
25.3±4.1
24.2±2.4
22.3±2.0
L5
16.0±4.7b
18.2±4.0b
23.2±6.4ab
42.6±13.4a
21.6±4.4ab
L6
94.0±23.3b
128.0±29.9ab 121.6±27.0ab 201.8±43.2a
L7
37.8±9.0b
52.5±11.8b
50.2±11.2b
87.3±17.9a
37.1±6.8b
A3
7.1±1.4ab
5.8±0.9b
8.2±1.0ab
5.7±1.1b
10.8±1.1a
A4
19.4±2.3
19.1±2.1
26.1±2.1
23.6±6.2
27.0±1.4
A5
9.5±2.3b
6. 8±1.3b
12.1±1.9ab
6.1±1.2b
19.1±1.9a
A7
28.4±3.9bc
26.3±3.4c
37.7±1.9ab
29.5±3.8abc
39.2±2.1a
A8
13.9±1.4c
15.4±0.9bc
20.3±1.2ab
15.8±1.5bc
21.9±1.4a
E1
17.7±1.9
18.5±3.3
22. 9±3.1
14.6±2.5
22.0±1.8
E2
34.5±4.9b
45.8±7.9ab
62.5±6.2a
50.7±7.4ab
61.4±3.7a
E3
5.2±0.9
6.4±1.0
7.7±1.2
6.0±1.1
8.9±1.4
C1c
Cb
CF
96.3±17.6b
Continued on next page.
103 Qian and Rimando; Flavor and Health Benefits of Small Fruits ACS Symposium Series; American Chemical Society: Washington, DC, 2010.
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Table 3. (Continued). Average contents of major compounds and the groups of volatiles (µgkg-1 FW equivalent of 3-octanol) in different origins of peach and nectarine cultivars Cla
Cb
CF
J
AE
E4
4.8±1.4
5.8±0.9
6.4±1.2
5.5±1.0
7.2±1.0
T3
19.5±9.8
11.5±2.9
23.8±6.5
21.3±9.5
35.3±5.7
T7
3.7±1.1b
3.9±0.5b
4.4±0.6b
4.1±0.8b
9.9±1.3a
T8
10.6±4.6ab
7.33±2.5b
11.7±2.8ab
8.3±1.8b
25.0±4.9a
T14
9.4±1.5
10.0±0.8
11.3±1.1
9.7±1.1
9.6±0.7
T16
6.6±1.2bc
11.5±1.4a
10.8±1.7ab
9.2±1.683abc 5.1±0.6c
K4
8.9±1.3
10.4±0.7
11.3±1.1
11.9±1.7
9.3±0.5
O1
7.0±0.9b
6.7±0.7b
11.8±1.7a
7.3±0.9b
11.4±0.5a
O2
29.4±10.1b
51.4±8.5a
43.2±5.2ab
40.2±9.3ab
25.2±3.8b
C6s
205.5±24.4b 258.2±38.9ab 313.2±30.9a
229.0±18.6ab 242.0±13.5ab
C6s % of TVd
26.80%
22.90%
Lactones
199.6±46.2b 245.0±51.1ab 230.7±47.3ab 381.1±76.9a
195.5±31.0b
Lactones% of TV
26.00%
Aldehydes
28.10%
30.70%
26.70%
26.10%
22.70%
38.10%
21.10%
103.1±12.7b 97.6±10.5b
130.0±9.8ab
96.3±13.3b
151.6±8.6a
Aldehydes% of TV
13.40%
10.60%
12.80%
9.60%
16.30%
Esters
64.5±8.7b
86.3±13.2ab
107.4±10.2a
89.0±10.9ab
103.7±5.7a
Esters% of TV
8.40%
9.40%
10.50%
8.90%
11.20%
Terpenoids
76.8±12.2
70.8±7.2
87.9±8.8
71.4±13.4
116.9±11.5
Terpenoids% of TV
10.00%
7.70%
8.60%
7.10%
12.60%
Alcohols
55.7±7.4b
67.2±5.1ab
67.7±4.3ab
76.9±12.0a
57.2±3.1b
Alcohols% of TV
7.30%
7.30%
6.60%
7.70%
6.20%
Ketones
12.9±1.9
14.3±0.9
15.1±1.5
15.2±2.1
14.3±0.9
Ketones% of TV
1.70%
1.60%
1.50%
1.50%
1.50%
Others
49.6±12.5b
79.0±10.7a
66.4±5.9ab
64.3±10.3ab
46.3±4.3b
Continued on next page.
104 Qian and Rimando; Flavor and Health Benefits of Small Fruits ACS Symposium Series; American Chemical Society: Washington, DC, 2010.
Table 3. (Continued). Average contents of major compounds and the groups of volatiles (µgkg-1 FW equivalent of 3-octanol) in different origins of peach and nectarine cultivars Cla
Cb
CF
J
AE
Others% of TV
6.50%
8.60%
6.50%
6.40%
5.00%
Total
767.7±57.0b 918.4±55.3ab 1018.5±68.7a 1000.3±70.7a 927.5±34.1ab
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a
Cl Chinese local cultivars; Cb pure Chinese original bred cultivars; CF China×foreign cultivars; J Japanese cultivars; AE American and European cultivars. b The different small letters indicate significant differences between populations (P