Postharvest Biology and Technology 108 (2015) 114119

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Postharvest Biology and Technology

journal homepage: www.elsevier.com/locate/postharvbio

Carotenoids and volatile proles of yellow- and red-eshed papaya

fruit in relation to the expression of carotenoid cleavage dioxygenase
genes
Guoxing Jinga,b , Taotao Lia , Hongxia Qua , Ze Yuna , Yongxia Jiaa , Xiaolin Zhengb ,
Yueming Jianga,*
a
Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou
510650, Peoples Republic of China
b
College of Food Science and Biotechnology, Zhejiang Gongshang University, Food Safety Key Laboratory of Zhejiang Province, Hangzhou 310035, Peoples
Republic of China

A R T I C L E I N F O

A B S T R A C T

Article history:
Received 11 January 2015
Received in revised form 6 June 2015
Accepted 7 June 2015
Available online xxx

Fruit of two different papaya cultivars (Sui huang, yellow esh, and Sui hong, red esh) were
characterized in terms of carotenoid and volatile proles throughout storage at 25
C. The results showed
that Sui huang cultivar exhibited signicantly lower carotenoid contents than Sui hong, which
accounted for the different color characteristic. The analyses of HPLC coupled to mass spectrometry
showed that zeaxanthin, b-cryptoxanthin and and b-carotene were identied as the most abundant
carotenoids in Sui huang fruit while the most abundant carotenoids were lycopene, followed by
b-carotene, zeaxanthin and b-cryptoxant hin in Sui hong fruit. The high expression of carotenoid
cleavage dioxygenase gene (CpCCD1) in Sui hong papaya fruit might stimulate the degradation of
b-carotene and lycopene and then form the specic volatiles of 6-methyl-5-hepten-2-one and b-ionone.
2015 Published by Elsevier B.V.

Key words:
Carica papaya
Carotenoids
Carotenoid cleavage dioxygenase
Mass spectrometry
Volatiles

1. Introduction
Papaya (Carica papaya L.) as a typically tropical fruit is a
commercially important fresh fruit crop in the international
market. The fruit have high contents of sugars, vitamin C and
carotenoids, and exhibit a pleasant aromatic avor (Bari et al.,
2006). Papaya fruit from various cultivars have different characteristics in color and avor. The color of papaya esh is linked to
contents of carotenoids (Wall, 2006). Carotenoids as lipid soluble
compounds play an important role in human health and nutrition.
Previous studies have shown that carotenoids prevent cardiovascular diseases by impacting cell signaling pathways (Stahl and Sies,
2005), in addition to protection against some types of cancer (Yuan
et al., 2003). Carotenoids are recognized also as strong antioxidant
compounds due to their abilities to trap singlet oxygen and
eliminate the peroxyl radical (Al Duais et al., 2009). Some
carotenoid compounds like a-carotene, b-carotene, g-carotene
and b-cryptoxanthin can reduce the risk of cancer and coronary
vein disease and inuence cell signaling pathways (Gayosso Garca

* Corresponding author. Fax: +86 20 37252831.

E-mail address: ymjiang@scbg.ac.cn (Y. Jiang).
http://dx.doi.org/10.1016/j.postharvbio.2015.06.006
0925-5214/ 2015 Published by Elsevier B.V.

Sancho et al., 2011; Stahl and Sies, 2005 and Yuan et al., 2003).
Furthermore, lycopene as a strong antioxidant activity may reduce
the incidence of cancer and cardiovascular diseases (Rao and
Agarwal, 2000). However, the contents and compositions of
carotenoids largely depend on the cultivars and/or production
regions of papaya. In these different cultivars, papaya fruit in red
esh color exhibits high nutritional benets because of the
accumulation of a large amount of lycopene (Schweiggert et al.,
2011).
Carotenoid compounds can be cleaved at their conjugated
double bonds by a carotenoid cleavage dioxygenase (CCD), forming
aldehydes and ketones which are important avor and fragrance
volatiles in many fruits (Paull et al., 2008). The recombinant
enzyme from Arabidopsis thaliana carotenoid cleavage dioxygenase
1 (AtCCD1) expressed highly in Escherichia coli can cleave the
multiple trans-carotenoid substrates (b-carotene, lutein, zeaxanthin and trans-violaxanthin) at the 9,10 and 90 ,100 -double bonds
and then produce a C14 dialdehyde and two C13 products (Schwartz,
2001). In tomato fruit, CCD1 generates the avor volatiles of
geranylacetone, pseudoionone, and b-ionone and, thus, the CCD1
alters the volatile compositions (Lewinsohn et al., 2005), resulting
in different volatile proles.

G. Jing et al. / Postharvest Biology and Technology 108 (2015) 114119

Carotenoids and aromatic odors have been identied as

important indicators for papaya commercial values. Sui hong
cultivar (red esh) has become a prominent variety because of high
carotenoid content and seductive volatiles, while Sui huang
cultivar (yellow esh) exhibits big morphological dimension and
high production but a weak odor. Up to now, the volatile
differences between red and yellow papaya esh was remain
undetermined, and the probably mechanism of resulting in
different volatiles of the various papaya was rather limited. The
objective of this study was conducted to compare levels of
carotenoids, then identify the major volatiles in two different
cultivars of Sui huang and Sui hong, and nally examine the
expression of carotenoid cleavage dioxygenase (CCD) gene in
relation to the volatile prole based on the breakdown of
b-carotene and lycopene. The study can help to understand better
the formation of the volatile compounds of papaya fruit during
storage.
2. Materials and methods
2.1. Fruit materials
Mature green fruit (L* value of 37.47  0.77, a* value of
14.95  0.4, b* value of 19.38  0.43, and rmness of about 12 N)
of papaya (Carica papaya L.) cv. Sui huang (yellow esh) and
Sui hong (red esh) were obtained from a commercial orchard in
Guangzhou, China. Fruit were selected for uniformity of shape
and weight, and then were washed. After air dried, 6 fruits were
placed in plastic boxes (50  80 cm) packed with 0.03 mm
polyethylene bags. In this experiment, 120 fruits (20 boxes with
6 fruits per box) from each cultivar were used. These fruits were
stored for 15 days at 25
C (ambient temperature) and 7595%
relative humidity (RH). Samples were taken after 0, 3, 6, 9, 12 and
15 days of storage.
2.2. Measurements of color characteristics
The esh color of 6 papaya fruit was determined using a Minolta
Chroma Meter CR-400 (Konica Minolta Sensing, Inc., Japan)
calibrated previously with a white standard tile by taking six
measurements per peeled fruit in the equatorial region (Schweiggert et al., 2011). The data were expressed as the color values of L*,
a*, b* and chroma (C*). Chroma (C*) indicating color intensity was
calculated by the formula (a*2 + b*2)1/2 while the hue angle (H
)
was calculated by the formula H
= arctan (b*/a*) (Schweiggert
et al., 2011), and the yellow index (YI) indicating the degree of
yellowness was calculated by the formula 142.86 b*/L* (Pathare
et al., 2012). For these determinations, 6 fruits were used.
2.3. Extraction and determination of total carotenoids
Total cartonoids were extracted according to the method of
Gayosso Garca Sancho et al. (2011) with some modications.
Flesh tissues (1 g) from 6 fruits of the two cultivars were
homogenized in 20 ml of hexane:dichloromethane (1:1, v/v)
containing 0.1 g/l butylated hydroxytoluene (BHT), and then
placed in a stirring bath set (HZQ-F160A, Shanghai, China) for 1 h
at 25
C in the dark. The extraction was then ltered through a
Whatman No. 1 paper (Whatman Inc., Shanghai, China) while reextraction of the recovered residue was carried out 2 times until a
colorless residue was obtained. For alkaline hydrolysis, 10 ml of
methanol:10% KOH (1:1, w/v) was added to the extract for 1 h at
50
C in a stirring bath set at 100 rpm. After saponication, the
upper fraction was dried with 5 g of Na2SO4. The contents of total
carotenoids were measured using a spectrophotometer (UVmini1240, Shimadzu Corp., Japan) at 450 nm. A calibration curve was

115

performed using b-carotene (Sigma, USA) in hexane as the

standard and hexane as the blank. The total carotenoid contents
were expressed as b-carotene equivalents on 100 g fresh weigh
(FW) basis. These determinations of total carotenoids were
repeated 3 times.
2.4. High-performance liquid chromatography ApCI-mass
spectrometry (HPLC-ApCI-MS) analysis
The carotenoid extract obtained above was evaporated with a
rotary evaporator (RE52AA, Yarong Equipment Co., Shanghai,
China) at 30
C. Samples were re-suspended in 2 ml methyl tertbutyl ether (MTBE) and then ltered through 0.45 mm PVD
membranes (Shanghai ANPEL Scientic Instruments Co., Ltd.,
Shanghai, China). Separation and qualitative analysis of carotenoid
compounds were performed using a high performance liquid
chromatography (HPLC) (Agilent, 1100HPLC, Germany) connected
to a Bruker Esquire 3000+ (Bruker, Bremen, Germany) equipped
with an atmospheric pressure chemical ionization (ApCI) interface
and Mass Hunter manager software (Version A. 02.01). The HPLC
system coupled with a 4.6  250 mm, 3 mm and C30 reverse phase
column (YMC Inc., Japan). The HPLC analysis was conducted by the
method of Schweiggert et al. (2011) with some modications. The
samples were eluted with a gradient system consisting of solvent A
(methyl tert-butyl ether, MTBE) and solvent B (methanol) as the
mobile phases, and the gradient elution was 0100% (B) within
55 min at a ow rate of 1 ml/min and 25
C. The injection volume
was 15 ml. The chromatogram was recorded in the range of 200
600 nm. Mass spectra of the major carotenoids were analyzed by
the method of Gayosso Garca Sancho et al. (2011) with some
modications. The ApCI-MS system was operated in the positive
and negative ion modes. High purity nitrogen (99.999%) was used
as a nebulizing (30 psi) and drying gas at a ow rate of 5 l/min.
Other ApCI-MS parameters consisted of the drying gas temperature of 300
C, vaporizer temperature of 400
C, corona of 10 mA,
capillary of 4 kV, fragmentor of 200 V, and skimmer voltages of
60 V. Carotenoids were identied by comparing their retention
times and UVvis data with those obtained with reference
standards and using their mass spectra (m/z 701000). In addition,
quantication for zeaxanthin, b-cryptoxanthin, b-carotene and
lycopene were performed by the HPLC analysis using calibration
curves of known standards (Sigma, USA). These analyses were
repeated three times.
2.5. Analyses of aroma volatiles
For volatile analysis, esh tissues from 6 papaya fruits of the
two cultivars were homogenized in a grinder (Philips, Shanghai,
China) for 30 s with an equal amount of 30% (w/v) NaCl, to inhibit
enzymatic degradation (Tietel et al., 2012). The homogenized
segments (100 g) were collected, and then placed in a 250 ml glass
reagent bottle. According the method of Tietel et al. (2012),
samples were held for 5 min at 40
C. Volatiles were collected for
additional 25 min from the vial headspace using a solid-phase
micro extraction with stable-ex bers (1 cm in length) coated
with a 50/30 mm layer of divinylben-zene/carboxen/ polydimethylsiloxane at 40
C. Gas chromatography (GC)/MS analysis was
performed according to the method of Wang et al. (2009) using a
GC-2010 gas chromatography (Shimadzu, Suzhou, China) equipped
with a GC MS-QP2010 plus mass spectrometer (Shimadzu, Suzhou,
China). A 30 m Rxi-5MS capillary column (0.25 mm i.d.) was used
for volatile separation while a split/splitless injector was used.
Samples (1 ml) were injected into the injector with a split ratio of
1:50. Oven temperature was kept for 3 min at 40
C, increasing to
120
C at a rate of 5
C/min and holding for 3 min, then increasing to
180
C at a rate of 2
C/min and holding for 3 min, and nally

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G. Jing et al. / Postharvest Biology and Technology 108 (2015) 114119

increasing to 230
C at a rate of 5
C/min and holding for 3 min.

Injector temperature was 250
C while the detector temperature
was 250
C, and the ion source temperature was 250
C. Helium
was used as the carrier gas. Identication of volatile compounds
was based on comparisons of their mass spectra with those
recorded in the National Institute of Standards and Technology
database and the peak area was used for measurement of volatile
content. The analysis was repeated three times.
2.6. RNA extraction and RT-PCR analysis
For CCD gene expression analysis, esh tissues (10 g) from 6
papaya fruits were taken after 0, 3, 6, 9, 12 or 15 days of storage and
then ground to a ne powder in the presence of liquid nitrogen.
Total RNA from papaya esh was extracted using the hot borate
method of Kuang et al. (2011). After the RNA extraction, potentially
contaminating DNA was eliminated by the treatment with DNase I
digestion using the RNase-free kit (TaKaRa, Dalian, China). The
total DNA-free RNA (100 ng) was used as template for reverse
transcription-PCR (RT-PCR), the synthesis of rst-strand cDNA
was performed by Reverse Transcription System (TaKaRa, Dalian,
China). The rst-strand cDNA of the product was subjected to PCR
amplication. Quantitative real-time was performed with a 7500

Fast Real-Time PCR System (Applied Biosystems, Foster City, CA,

USA) using SYBRPremix Ex TaqTM mix (TaKaRa, Dalian, China)
and the uorescence data was analyzed using 7500 Fast System
SDS Software 2.0.1. RT-PCR was carried out with 20 ng cDNA by
adding 10.0 ml SYBRPremix Ex TaqTM, 0.5 ml PCR forward primer
(10 mM), 0.5 ml PCR reverse primer (10 mM) and 0.5 ml ROX
reference dye II. The RT-PCR procedure consisted of 3 min at
95
C, followed by 40 cycles at 95
C for 5 s, 55
C for 5 s and 72
C
for 20 s. In this study, the most stable reference gene 18S rRNA
(Devitt et al., 2009) was used as the internal control. The primer
pairs for CCD1, CCD7 and 18S rRNA were provided in Supplementary Table S1. The relative expression levels of target genes were
calculated with formula 2DDCT (Kuang et al., 2011). Values
represented the averages and standard error (SE) of three
biological replicates.
2.7. Data handling
The experiments were arranged in completely randomized
design, and each measurement was comprised of at least three
replicates. Data were analyzed by analysis of variance using SPSS
version 7.5. Least signicant differences (LSD) were used to
compare signicant effects at the 5% level.

Fig. 1. Changes in the a*, b*, L*, H* and C* values, and yellow index (YI) in esh of Sui huang and Sui hong papaya fruit during storage. Each value is presented as
mean  standard error (n = 3). The vertical bars indicate standard errors where they exceeded the symbol size (*, Sui huang; , Sui hong). Asterisk indicates a signicant
(P < 0.05) difference between the Sui huang and Sui hong papaya fruit.

G. Jing et al. / Postharvest Biology and Technology 108 (2015) 114119

117

Yamamoto (1964) reported lycopene, b-cryptoxanthin and

b-carotene present in the red papaya esh. Rivera Pastrana et al.
(2010) and De Souza et al. (2008) identied lycopene, b-cryptoxanthin and b-carotene as the major carotenoids in papaya esh.
Besides the presence of b-cryptoxanthin, b-carotene and lyco-

3. Results and discussion

3.1. Color characteristic
As shown in Fig. 1, signicant differences in color parameters L*,
a*, b*, C*, H
and YI between the two cultivars were observed.
During fruit storage, the Sui hong papaya esh (19.733.5)
exhibited higher a* values than Sui huang (4.4110.09). In
contrast, the L* and H
values of Sui hong were signicantly lower
than that those of Sui huang. The b*, C* and YI values of the two
cultivars of papaya fruit had similar changes during storage.
The parameter a* represents reddish color while the parameter
b* and yellowness index (YI) indicate yellow color and yellow
degree, respectively, and chroma (C*) is used to estimate the
degree of difference of a hue in comparison to a grey color with the
same lightness. The higher the chroma value, the higher the color
intensity of samples perceived by human (Pathare et al., 2012). This
present study showed that the b*, C* and YI values may largely
affected by the increase of yellow pigments, which was in
agreement with the results of De Souza et al. (2008), who reported
the esh color of Golden papaya was less intensity than Tainung
01 hybridand, which may be due to its lower content in lycopene.
The L* value is regarded as an approximate measurement of
luminosity while the hue angle (H
) value represents lesser yellow
character. In this study, the decrease in the L* and H
values of esh
color could be related to the increase in the content of the total
carotenoids (Table 1), which was in agreement with the changes in
the L* and H
values reported by Schweiggert et al. (2011).

pene, the results exhibited that zeaxanthin was the fourth major
carotenoid in the Sui hong papaya esh.
3.3. Content of carotenoids
As shown in Table 1, the contents of total carotenoids of papaya
fruit increased to the maximum values after 12 days of storage,
and then declined. The contents of the total carotenoids in Sui
hong were always higher than Sui huang, b-carotene content
reached the maximum value of 114.63 mg/100 g FW while the
b-cryptoxanthin content had the maximum value of approximately 50 mg/100 g FW after 15 days of storage, and the
zeaxanthin content increased from 17.52 to 87.88 mg/100 g FW
in Sui huang papaya fruit. In comparison with Sui huang, the
content of lycopene as the major carotenoid increased from
539.59 mg/100 g FW to the maximum value of 1863.23 mg/100 g
FW in Sui hong fruit after 12 days of storage. It was noted
particularly that the b-cryptoxanthin content of the Sui hong
papaya fruit increased rapidly from 0.33 to 71.4 mg/100 g FW
during storage. However, b-carotene as the second most
abundant carotenoid and its content increased slowly from
26.64 to 161.30 mg/100 g FW, followed by zeaxanthin which
increased 23.0173.68 mg/100 g FW.
Previous studies demonstrated that the color of mango and
papaya fruits can be characterized well by the content of the total
carotenoids, which played an important role in the fruit
acceptability by consumers (Gayosso Garca Sancho et al., 2011).
In the present study, the total carotenoid content of Sui huang
were higher than those reported for the yellow esh cultivars such
as Kapoho, Laie gold and Rainbow (Wall, 2006), while the Sui
hong cultivar exhibited higher carotenoid contents than those
reported for the red esh cultivars such as Formosa, Sunrise and
Maradol (Melo et al., 2006), with carotenoid content ranging from
1.77 to 6.21 mg/100 g FW as described by De Souza et al. (2008),
Gayosso Garca Sancho et al. (2011) and Schweiggert et al. (2011).
These differences in the carotenoid content accounted for the
different color characteristic between the two cultivars in this
study (Fig. 1). Moreover, the lycopene content in Sui hong papaya
fruit was comparable to those reported in other red esh papaya
cultivars ranging between 1200 and 3500 mg/100 g FW by De
Souza et al. (2008), Gayosso Garca Sancho et al. (2011) and
Schweiggert et al. (2011). However, about 4.7 mg/100 g FW in

3.2. Identication of carotenoid compounds

Four major carotenoid compounds were separated and identied in the Sui hong papaya esh (Supplementary Fig. S1 and
Table S2). The identication of four carotenoid compounds were as
follows: peak 1 corresponding to zeaxanthin with a predominant
ion [M + H]+ at 569.5 and yielding ion fragments at m/z 551.4, 533.4,
477.4 and 459.4 (Supplementary Fig. S2); peak 2 corresponding to
b-cryptoxanthin [M  H]+ at 553.5 and yielding ion fragments at
m/z 535.5, 518.5 and 468.5 (Supplementary Fig. S3); peak 3
corresponding to b-carotene [M  H] at 536.8 and yielding ion
fragments at m/z 518.2, 433.4 and 401.4 (Supplementary Fig. S4);
and peak 4 corresponding to lycopene [M + H]+ at 537.5 and
yielding ion fragments at m/z 519.5, 481.4, 467.4, 455.4 and 427.3
(Supplementary Fig. S5). In comparison with the Sui hong, three
carotenoid compounds of zeaxanthin, b-cryptoxanthin and
b-carotene were identied but no lycopene was detected in the
Sui huang cultivar (Table 1).

Table 1
The contents of the major carotenoids in esh of Sui huang and Sui hong papaya fruit during storage. Each value is presented as mean  standard error (n = 3). ND, no
detection.
Cultivars

Carotenoids
(mg/100 g FW)

Sui huang

Zeaxanthin
b-cryptoxanthin
b-carotene
Lycopene
Total carotenoids

17.52  4.7
0.68  0.04*
17.09  3.3
ND
208.67  4.62

30.99  6.2
10.29  1.4*
27.32  4.7

49.39  6.8*
12.91  1.7
34.09  4.5

54.75  4.8
16.97  2.9
47.47  6.1

77.92  6.4
44.80  5.8
80.65  6.5

87.88  10.1*
50.07  4.2
114.63  9.2

486.37  15.03

981.83  20.83

1778.54  95.38

2188. 33 110.55

2166.83  129.79

Zeaxanthin
b-cryptoxanthin
b-carotene
Lycopene
Total carotenoids

23.01  1.8
0.33  0.006
26.64  4.1*
539.59  44.2
874.67  5.81*

32.67  3.0
2.92  0.03
47.13  3.0*
1031.3  95.3
2540.75  23.44*

60.39  2.9*
43.72  3.1*
88.76  6.8*
1460.34  125.6
3186.65  129.49*

72.78  7.5
65.24  4.8*
139.91  11.2*
1863.23  237.8
4594.36  278.4*

73.68  3.9
71.4  6.1*
161.3  8.7*
1621.79  216.0
4534.26  182.92*

Days of storage
0

Sui hong

35.84  4.2
14.91  2.0
50.75  4.1*
1148.55  102.9
2864.24  18.9*

12

Represents a signicant (P < 0.05) difference in carotenoid content between Sui huang and Sui hong cultivars.

15

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G. Jing et al. / Postharvest Biology and Technology 108 (2015) 114119

Table 2
Relative content (%) of the volatiles of papaya fruit during storage.
Cultivars

Retention time
(min)

Volatiles

Relative content (%) of volatiles of papaya fruit at various storage day

0

12

15

Sui huang

10.100
22.750
10.100
22.750

6-Methyl-5-hepten-2-one
Trans-b-ionone
6-Methyl-5-hepten-2-one
Trans-b-ionone

0
0
0
0

0
0
0.48  0.09*
1.81  0.22*

0
0
2.21  0.18*
1.79  0.20*

0
0
2.49  0.03*
2.69  0.25*

0
0
3.43  0.30*
3.45  0.21*

0
0
3.54  0.12*
3.05  0.08*

Sui hong
*

Represents a signicant difference (P < 0.05) in the two volatiles between Sui huang and Sui hong cultivars.

yellow esh cultivar and about 60 mg/100 g FW of zeaxanthin in red

esh cultivar were reported (Barreto et al., 2011; Murillo et al.,
2010). Thus, the different varieties, production area, sunlight
exposure, stage of ripeness and methodology used for analysis
might result in the differences in the contents of the four major
carotenoids (Gayosso Garca Sancho et al., 2011).
3.4. Identication of volatile compounds
A major volatile prole of papaya esh of two cultivars is
presented in Supplementary Table S3. Methyl butanoate, ethyl
butanoate, D-limonene, 5-ethenyltetrahydro-a, a,5-trimethyltrans-2-furanmethanol, b-linalool, ethyl octanoate, methyl geranate, cis-gerany lacetone, methyl tetradecanoate, ethyl myristate
and ethyl 9-hexadecenoate were all detected in the two cultivars of
papaya, and b-linalool was the most abundant component (35.44%
and 54.56%, respectively) in the volatile proles of Sui huang and
Sui hong papaya (Supplementary Table S3). Acetone, hexane,
trichloromethane and dimethylsilanediol were the specically
small molecule volatiles in Sui huang cultivar. In contrast, the
small molecule volatiles such as ethyl alcohol, dimethyl sulde and
acetic acid existed typically in Sui hong cultivar. Furthermore, the
specically long chain esters of ethyl (9E)-9-octadecenoate, ethyl
octadecanoate, methyl a-linolenate and methyl g-linolenate were
only detected in Sui huang cultivar. It was particularly noted that
6-methyl-5-hepten-2-one and b-ionone existed specically in Sui
hong fruit during storage (Supplementary Fig. S6), and the UVvis
spectra and mass ion fragments of the two volatiles were in
agreement with these reports in Zelena et al. (2009) and Vogel
et al. (2008). At the beginning of storage, the two volatiles were not
detected in Sui hong esh, but their relative contents measured by
GC/MS increased with storage time. The relative contents of 6methyl-5-hepten-2-one and b-ionone reached the maximum
values after 12 and 15 days of storage, respectively (Table 2).

Lewinsohn et al. (2005) reported that the carotenoid compositions correlated with the components of the monoterpene and
norisoprenoid volatiles. Color and aroma compounds are highly
associated in carotenoid compositions. Watermelon fruit and wild
type (red) tomatoes contained high lycopene and low concentrations of b-carotene, and they accumulated non-cyclic norisoprenoids such as 6-methyl-5-hepten-2-one, farnesyl acetone, (E,
E)-pseudoionone, 2,3-epoxygeranial, 2,6-dimethyl hept-5-1-al,
geranyl acetone, and dihydro-apo-farnesal in addition to geranial
and neral, and the cyclic norisoprenoid b-ionone (Lewinsohn et al.,
2005), but the orange-colored B genotype accumulated more
b-carotene in addition to lycopene, and produced the volatiles of
dihydroactinodiolide, b-ionone, and b-cyclocitral. In this study,
the Sui hong papaya contained high contents of b-carotene and
lycopene which could generate 6-methyl-5-heptene-2-one and
b-ionone, but the two volatiles were not detected in Sui huang
papaya. The other potential volatiles such as b-ionone-5,6epoxide, dihydroactinidiolide, b-cyclocitral and 2-hydroxy-2,6,6trimethylcyclohexanone degraded from b-carotene present in
tomato and watermelon fruits were not detected in Sui huang and
Sui hong papaya fruits, and the non-cyclic volatile of pseudoionone could not be degraded from lycopene.
3.5. Expression of CpCCD1 and CpCCD7
According to papaya genome information from http://www.
phytozome.net, we retrieved two CCD genes (evm.model.supercontig_90.25 and evm.TU.supercontig_125.7) related to the
degradation of lycopene and b-carotene (Supplementary
Table S1). The expressions of CpCCD1 and CpCCD7 of esh tissues
of the two cultivars of papaya fruit during storage are presented in
Fig. 2. The initial expression of CpCCD1 in Sui hong fruit was
signicantly (P < 0.05) lower than that in Sui huang, but it
increased rapidly after 3 days of storage. Although the gene

Fig. 2. Changes in the relative expressions of CpCCD1 and CpCCD7 in esh of Sui huang and Sui hong papaya fruit during storage. Each value is presented as mean  standard
error (n = 3). The vertical bars indicate standard errors where they exceeded the symbol size (*, Sui huang; , Sui hong). Asterisk indicates a signicant (P < 0.05) difference
between the Sui huang and Sui hong papaya fruit.

G. Jing et al. / Postharvest Biology and Technology 108 (2015) 114119

expression in Sui hong fruit decreased after 6 days of storage, it

was signicantly (P < 0.05) higher than that in Sui huang. In
contrast, the expression of CpCCD7 exhibited a lower level
compared with that of CpCCD1 in the two cultivars of papaya
fruits throughout the storage period.
Carotenoid cleavage can occur at any conjugated double bonds
by CCD to form an aldehyde or ketone, or the CCDs in Arabidopsis
directly involved in the generation of volatiles and non-volatile
apocarotenoids (Vogel et al., 2008). The formation of b-ionone
resulted from the cleavage of b-carotene at the 9,10 and 90 ,100
double bonds while 6-methyl-5-heptene-2-one was generated by
lycopene cleavage at 5, 6 and 50 ,60 double bonds (Vogel et al., 2008).
Simkin et al. (2004) reported that LeCCD1A and LeCCD1B in tomato
fruit can generate the aldehydes and ketones including 6-methyl5-hepten-2-one, geranyl acetone, pseudoionone and b-ionone,
while maize CCD1 (ZmCCD1) can cleave linear and cyclic
carotenoids and produce 6-methyl-5-hepten-2-one by the cleavage of 5,6 or 50 ,60 bond positions of lycopene and b-ionone derived
from 9,10 or 90 ,100 bond cleavage of b-carotene (Vogel et al., 2008).
In this study, lycopene and b-carotene might be degraded with
the enhanced expression of CpCCD1, and could form the non-cyclic
and cyclic special volatiles of 6-methyl-5-hepten-2-one and
b-ionone in Sui hong fruit (Supplementary Fig. S7). Although a
high content of b-carotene existed in Sui huang papaya fruit, the
low expression of CpCCD1 and CpCCD7 may not ensure to generate
b-ionone which was not be detected by GC/MS.
4. Conclusions
Zeaxanthin, b-cryptoxanthin and b-carotene were identied as
the major carotenoids in Sui huang papaya esh whereas the most
abundant carotenoid was found to be lycopene, followed by
b-carotene, zeaxanthin and b-cryptoxanthin in Sui hong esh.
The signicant difference in lycopene content can account for the
formation of red esh in Sui hong papaya fruit. The volatiles of 6methyl-5-hepten-2-one and b-ionone specically existed in Sui
hong fruit, but they were not detected in Sui huang fruit.
Furthermore, the high expression of CpCCD1 in Sui hong papaya
fruit might stimulate the degradation of b-carotene and lycopene
and then form the pleasant aromatic odor.
Acknowledgements
This work was supported by the National Natural Science
Foundation of China (grant no. 31271971), the Planning Project of
Chinese Academy of Sciences (grant no. KSZD-EW-Z-021-3-3) and
Guangdong Province Group Team for Equipment Technology of
High Efciency Drying and Cold Chain Transport of Agricultural
Products.
Appendix A. Supplementary data
Supplementary data associated with this article can
be found, in the online version, at http://dx.doi.org/10.1016/j.
postharvbio.2015.06.006.
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