Effect of Ozone on Postharvest Quality of Persimmon

<strong>Effect</strong> <strong>of</strong> <strong>Ozone</strong> on Postharvest
Quality <strong>of</strong> Persimmon
ALEJANDRA SALVADOR, ISABEL ABAD, LUCíA ARNAL, AND J.M. MARTíNEZ-JÁVEGA
ABSTRACT: The effect <strong>of</strong> gaseous ozone exposure on the quality <strong>of</strong> persimmon picked at 2 different harvest dates
was evaluated. Fruit from both harvests were continuously exposed to 0.15 ppm (vol/vol) <strong>of</strong> ozone for 30 d at 15 ◦ C,
90% relative humidity (RH). Then, fruit were submitted to astringency removal treatment (24 h at 20 ◦ C, 98% CO2)
and stored for 7 d at 20 ◦ C (90% RH) in order to simulate shelf-life period. The most important disorder was flesh
s<strong>of</strong>tening, which took place when fruit were transferred from 15 ◦ C to shelf-life conditions. In the 2nd harvest, where
the fruit were harvested with lower firmness, ozone maintained firmness over commercial limits even after 30 d at
15 ◦ C plus shelf-life. <strong>Ozone</strong>-treated fruit showed the highest values <strong>of</strong> weight loss, coinciding with the maximum
electrolyte leakage (EL) percentage. <strong>Ozone</strong> did not affect color index (CI), ethanol, total soluble solids (TSS), or pH.
Unremarkable differences in acetaldehyde were observed between fruit submitted to ozone treatment and control
fruit. No phytotoxic injuries in tissues were observed in ozone-treated fruit.
Keywords: firmness, harvest date, ozone, persimmon, storage
Introduction
Persimmon crop has undergone an important expansion in the
last years, becoming a good alternative to the other fruit crops in
the Mediterranean area. In this way, from 1992 to 2003, persimmon
crop has increased close to 150%. The main reason for this expansion
is the new way to present “Rojo Brillante” cultivar, without astringency
and with high firmness after being submitted to astringency
removal treatment using high concentration <strong>of</strong> CO2 (Arnal and Del
Río 2003). This fact has allowed opening <strong>of</strong> new market perspectives,
<strong>of</strong>fering advantages over standard traditional persimmon, s<strong>of</strong>t fruit
with very difficult postharvest manipulation.
A point to note is that persimmon production in this area is centralized
in this cultivar, whose harvest period is short, from October
to December. This is because one <strong>of</strong> the main goals is to prolong
postharvest life. Fruits <strong>of</strong> cultivar “Rojo Brillante” are sensitive to
low temperatures and show a drastic s<strong>of</strong>tening as the manifestation
<strong>of</strong> chilling injury. As consequence, fruit storage is carried out close to
15 ◦C, nevertheless this temperature is too high to prolong storage
(Arnal and Del Río 2004; Salvador and others 2005). On the other
hand, although persimmon has not high susceptibility to pathogen
contamination, this storage temperature does not help to fungal
control.
The effect <strong>of</strong> ozone exposure on fungal decay during postharvest
storage has been studied for different commodities with some discrepancies
in the results. <strong>Ozone</strong> does not seem to control pathogen
in wounds, and cannot be substituted for the synthetic fungicides
that are currently applied in packing-lines; nevertheless exposure
to gaseous ozone inhibits the aerial growth <strong>of</strong> the mycelia and prevents
sporulation <strong>of</strong> the important postharvest pathogens in different
commodities (Palou and others 2001, 2002, 2003).
The benefits <strong>of</strong> ozone depend on different factors, such as environmental
conditions, temperature, and humidity (Palou and others
MS 20060019 Submitted 1/11/2006, Accepted 4/24/2006. The authors are with
Centro de Tecnología de Postcosecha, Inst. Valenciano de Investigaciones
Agrarias, 46113 Valencia, Spain. Direct inquiries to author Salvador (E-mail:
asalvado@ivia.es).
Further reproduction without permission is prohibited
2001). Due to the lack <strong>of</strong> ozone penetration, the efficacy <strong>of</strong> this treatment
also depends on the type <strong>of</strong> package; therefore, no benefits
were found for ozone gas exposure when citrus fruit were packed
in corrugated fiberboard cartons or plastic bags (Palou and others
2003). On the other hand, the ability <strong>of</strong> gaseous ozone to react
with ethylene leads to this treatment as potential tool to be used
in rooms where sensitive ethylene commodities were stored (Dickson
and others 1992; Skog and Chu 2001). In this sense, persimmon
fruit normally produce small amounts <strong>of</strong> ethylene; however, it is
known that marked increases in the endogenous levels <strong>of</strong> ethylene
are produced by different conditions, such as wounds or pathogen
presence. Additionally, persimmon fruit is very sensitive to ethylene
action, and exposure to low concentrations <strong>of</strong> ethylene accelerates
s<strong>of</strong>tening (Nakano and others 2003).
<strong>Ozone</strong> treatment may also affect quality parameters. Skog and
Chu (2001) reported that ozone (0.4 μLL- 1 ) improved the quality <strong>of</strong>
broccoli and cucumber stored at 3 ◦ C, prolonging the storage life.
Nevertheless, no effects <strong>of</strong> ozone treatment were found on quality
parameters on cucumbers stored at 10 ◦ C, mushrooms stored
at 4 ◦ C, or apples and pears kept at 1 ◦ C. In strawberry, ozone induced
delay <strong>of</strong> fruit s<strong>of</strong>tening and weight loss; however it increased
a loss <strong>of</strong> fruit aroma (Perez and others 1999; Nadas and others 2003).
<strong>Ozone</strong> storage also resulted in market quality extension <strong>of</strong> blackberries
(Barth and others 1995). Treatments with ozonated water
also have been effective in reducing microbial population and in
improving the storage quality on fresh-cut celery, cilantro, and lettuce
(García and others 2003; Wang and others 2004; Zhang and
others 2005). However, immersion in ozonated water did not control
postharvest decay <strong>of</strong> citrus fruit, peaches and nectarines, and
table grapes (Smilanick and others 2002).
The effects <strong>of</strong> ozone exposure on fruit and vegetable quality seem
to be highly dependent on the commodities, and therefore must be
individually assessed for each commodity at its ideal storage temperature
(Liew and Prange 1994).
Storage under atmosphere containing ozone is being considered
an alternative to maintain persimmon at 15 ◦ C, but there is no clear
information about the effects on quality parameters. In this study,
S: Sensory & Nutritive Qualities <strong>of</strong> Food

S: Sensory & Nutritive Qualities <strong>of</strong> Food
<strong>Ozone</strong> effect on persimmon . . .
the effect <strong>of</strong> ozone treatment on the quality <strong>of</strong> persimmon picked at
2 different harvest dates, during storage at 15 ◦ C, was evaluated.
Materials and Methods
Plant material and treatments
“Rojo Brillante” persimmons (Diospyros kaki L.) were obtained
from L’Alcudia (Valencia, Spain) at 2 harvest dates, on October 27
(Harvest 1) and on November 24 (Harvest 2). Fruit were transported
immediately to the Inst. Valenciano de Investigaciones Agrarias
(IVIA) experimental station, where they were carefully selected for
uniformity <strong>of</strong> size and color development and randomly separated
into 2 groups by personnel familiar with this persimmon cultivar.
Both groups were stored in chambers <strong>of</strong> 1.5 m 3 for up to 30 d at
15 ◦ C, 90% RH, one in air (control) and the other in air enriched
with 0.15 ppm <strong>of</strong> ozone. An ozone generator (mod XES-030, Grupo
Interozone, Santiago, Chile) was used to achieve the desired concentration.
After 10, 20, and 30 d at 15 ◦ C, fruit were submitted to
astringency removal treatment (24 h at 20 ◦ C, 98% CO2) (Salvador
and others 2004) and then held for 7 d at 20 ◦ C, 90% RH to simulate
shelf-life. Astringency removal was carried out in closed containers,
and the desired atmosphere was established by passing a stream <strong>of</strong>
air containing 98% CO2 through the containers.
<strong>Ozone</strong> concentration was continuously monitored by UV absorption
ozone analyzer (Model IN-2000-1, INUSA Inc., Needham, Mass.,
U.S.A.) with a minimum detection limit <strong>of</strong> 0.01 ppm. Air from the
ozonated chamber was pumped through a Teflon tube to the analyzer,
which was located in an adjacent room. <strong>Ozone</strong> level was also
periodically measured using an ozone detector tube (0.05 to 0.7 ppm,
nr 6733181, Dräger Accuro, Canada Ltd., Mississauga, Ont., Canada)
Fruit quality assessment
Flesh firmness was determined over 20 fruit per treatment with a
Texturometer Instron Universal Machine model 4301 (Instron Corp.,
Canton, Mass., U.S.A.), using an 8-mm plunger. Results were expressed
as load in Newtons (N) to break the flesh in each fruit on
opposite sides after peel removal.
Weight loss was measured on 20 fruit per treatment, with the results
expressed as a percentage. Electrolyte leakage (EL) was determined
in triplicate for each sample. Six-mm dia pericarp discs were
excised from the equatorial region <strong>of</strong> the fruit. Three lots <strong>of</strong> discs,
approximately 2 g, were rinsed 3 times in distilled water, placed
in 100 mL <strong>of</strong> 0.4 M mannitol solution, and incubated for 3 h at
room temperature in continuous shaking. The electrical conductivity
readings <strong>of</strong> the solution were taken at 30-min intervals as a
measure <strong>of</strong> EL from the discs, using a conductance meter (model
Consort C231, Electrochemical Multimeter, Belgium). After 3 h, the
flasks were autoclaved for 30 min at 121 ◦ C and cooled, and a final
conductivity reading was taken for total electrolytes. EL was expressed
as a percentage <strong>of</strong> the total electrolytes.
Skin color was evaluated by a Minolta Colorimeter (model CR-
300, Minolta Co. Ltd., Osaka, Japan) on samples <strong>of</strong> 20 fruit per treatment.
L, a, and b Hunter parameters were measured and results were
expressed as skin color index (CI) (Jiménez-Cuesta and others 1981).
Color index = (1000a)/(Lb)
Three samples <strong>of</strong> juice sample from 15 different fruit per treatment
were used to analyze total soluble solids (TSS), ethanol, and acetaldehyde
concentrations. TSS were measured with a digital refractometer
(model PR-1, Atago, Japan) and expressed as ◦ Brix. To
measure ethanol and acetaldehyde concentrations, 5 mL <strong>of</strong> the filtered
juice were transferred to 10 mL vials with crimp-top caps and
a TFE/silicone septa seal and frozen (−20 ◦ C) until analysis. Thereafter,
samples were put in a water bath at 20 ◦ C for 1 h and then
heated at 60 ◦ C for 10 min in a water bath. One milliliter <strong>of</strong> the
headspace gas was withdrawn from the vials and injected into a gas
chromatograph Perkin-Elmer (model 1020, Perkin Elmer Corp., Norwalk,
Conn., U.S.A.), provided with a flame ionization detector (FID)
and a 1/8 ′ × 1.2 m Poropak QS 80/100 column. The injector was set
at 175 ◦ C, the column at 150 ◦ C, the detector (FID) at 200 ◦ C, and the
carrier gas at 12.3 psi. Acetaldehyde and ethanol were identified by
comparison <strong>of</strong> retention times with standards and were expressed
as mg/100 mL.
Data were subjected to analysis <strong>of</strong> variance (ANOVA), and least
significant difference (LSD) at the 5% level was used for comparing
means using the Statgraphics plus 2.1 (Manugistics, Inc., Rockville,
Md., U.S.A.).
Results and Discussion
Firmness is considered one <strong>of</strong> the most important quality parameters
in persimmon fruit when it is commercialized after
astringency removal. From previous experiments in this cultivar,
firmness values lower than 10 N force have been considered as not
acceptable from a commercial point <strong>of</strong> view (Salvador and others
2004).
In the present study, in both harvest dates, a gradual decrease <strong>of</strong>
firmness fruit was observed throughout storage at 15 ◦ C (Figure 1),
this s<strong>of</strong>tening being more marked after deastringency treatment
plus shelf-life. In Harvest 1, firmness loss was similar in control fruit
than in ozone-treated fruit throughout the whole experiment. In
Harvest 2, fruit were harvested with lower firmness and differences
between treatments were not observed during storage at 15 ◦ C; nevertheless
after shelf-life conditions, nontreated fruit underwent a
more important s<strong>of</strong>tening than ozonated fruit. In this way, after
10 d at 15 ◦ C plus shelf-life, control fruit presented noncommercial
Firmness (N)
Firmness (N)
60
50
40
30
20
10
0
60
50
40
30
20
10
0
c
c
c
bc
b
b
b
b
a
a
Control <strong>Ozone</strong>
a
10 d 20 d 30 d 10d 20d 30d
days at 15∫C 15°C days at 15∫C+5 15°C+5 d. at 20∫C 20°C
a
10 d 20 d 30 d 10d 20d 30d
c
a
days at 15∫C 15°C days at 15∫C+5 15°C+5 d. at 20∫C 20°C
c
b
b
a
b
b
a
a
H1
a
H2
Figure 1 --- <strong>Effect</strong> <strong>of</strong> ozone treatment on firmness during
storage at 15 ◦ C and after subsequent shelf-life period at
20 ◦ C (90% RH) <strong>of</strong> “Rojo Brillante” persimmon harvested
at 2 different dates: Harvest 1 (H1) and Harvest 2 (H2).
Fruit were submitted to astringency removal treatment
prior to transfer to 20 ◦ C. Means for storage at 15 ◦ C or for
shelf-life periods with the same letter are not significantly
different at 5% level (LSD test).
b

<strong>Ozone</strong> effect on persimmon . . .
values; meanwhile treated fruit showed values slightly superior to
the commercial limit even after 30 d plus shelf-life.
In previous studies, higher firmness was found in citrus and cucumbers
treated with ozone compared with control (Nadas and others
2000; Skog and Chu 2001). <strong>Ozone</strong> also induced a delay <strong>of</strong> fruit
s<strong>of</strong>tening in strawberry during cold storage and storage at room
temperature (Nadas and others 2003).
Weight loss is not usually a limiting factor in persimmon postharvest
life due to their skin, which avoids water losses. In both studied
harvests, similar weight losses were shown (Table 1), finding
the most important values after simulating shelf-life. <strong>Ozone</strong>-treated
fruit showed the highest weight loss, achieving values <strong>of</strong> 3.6% after
30 d <strong>of</strong> storage plus shelf-life.
Crisosto and Palou (personal communication) also observed
higher weight loss in peaches treated with ozone than in nontreated
ones; the highest water loss showed by the treated fruit was related
to cuticle damage caused by ozone.
In our research, EL was measured in assessment carried out with
early harvested fruit (Figure 2). The highest EL showed in ozonetreated
fruit coincided with the highest weight loss observed in this
fruit. According to Rao and others (2000), it seems that ozone can react
with cuticular components and with lipids and proteins in plants,
as result suggests that membrane lipids are susceptible to ozone
damage. In cucumber, although ozone treatment showed positive
effects during storage, the ozone samples appeared more desiccated
than the control (Skog and Chu 2001). With ozone residual concentration
<strong>of</strong> 10 to 22 μL L-1 at 2 ◦ C, Liew and Prange (1994) observed
Table 1 --- <strong>Effect</strong> <strong>of</strong> ozone treatment on weight loss during
storage at 15 ◦ C and after subsequent shelf-life period
at 20 ◦ C (90% RH) <strong>of</strong> “Rojo Brillante” persimmon harvested
at 2 different dates: Harvest 1 and Harvest 2. Fruit
were submitted to astringency removal treatment prior to
transferring fruit to 20 ◦ C.
Weight loss (%)
Days at 15 ◦C Daysat15◦C + 5dat20◦C 10 20 30 10 20 30
Harvest 1
Control 0.4a 0.6b 0.8c 2.3b 1.8a 1.9a <strong>Ozone</strong> 1.4d 1.7e 2.4f 3.5d 2.9c 3.6d Harvest 2
Control 0.2a 0.3a 0.5b 0.2a 1.3b 2.0c <strong>Ozone</strong> 0.9c 1.6d 2.1f 1.4b 2.8d 3.5e a–f Data followed by the same letter for storage at 15 ◦ C or for shelf-life periods
did not differ at 5% significance level.
Electrolyte leakage (%)
40
35
30
25
20
15
10
Harvest 1 Control <strong>Ozone</strong>
5
20 days at 15°C + 5 d. at 20°C
0
0 30 60 90 120 150 180
Electrolyte leakage (%)
40
35
30
25
20
15
10
5
0
30 days at 15°C + 5 d. at 20°C
0 30 60 90 120 150 180
Incubation Time (minutes) Incubation Time (minutes)
symptoms <strong>of</strong> physiological disruptions, such as respiration rate, EL,
and color changes. In other study, browning observed in the cut surface
in broccoli stored with ozone (1.7 μL L- 1 ), to also related to an
increase <strong>of</strong> membrane permeability and a loss <strong>of</strong> cellular compartimentation
(Skog and Chu 2001).
In contrast to our results, in strawberry, a reduction <strong>of</strong> weight
loss was observed with ozone treatment that was related to a better
maintenance <strong>of</strong> fruit firmness (Nadas and others 2003), and in
carrots, ozone did not affect weight loss (Liew and Prange 1994).
Fruit from Harvest 1, with a very low external coloration, gradually
increased the CI during storage at 15 ◦ C, and this increase was
more accentuated after astringency removal treatment and subsequent
shelf-life (Figure 3). In Harvest 2, the initial CI was higher
and slight increases in coloration occurred during storage at 15 ◦ C
until achieving the maximum CI values, close to 30, without significant
increment after transferring the fruit at shelf-life conditions.
No differences were observed between ozone-treated fruit and nontreated
fruit in both harvests. In other commodities, color changes
have been affected by the ozone treatment. Nadas and others (2000)
observed that treatment with ozone delayed color evolution in citrus
fruit. <strong>Ozone</strong> also affected the carrot color (Liew and Prange 1994).
Acetaldehyde production was very low at both harvests and slight
changes were shown during storage at 15 ◦ C (Table 2). After astringency
removal treatment plus 5 d at 20 ◦ C, this volatile increased due
to anaerobic conditions induced by CO2 treatment (Arnal and Del
Río 2003). Unremarkable differences were observed between fruit
submitted to ozone atmosphere and control fruit. <strong>Ozone</strong> had no effect
on ethanol production throughout the assay (data not shown).
At harvest, fruit from H1 presented lower TSS values than fruit
from H2, and no changes were observed throughout storage at 15 ◦ C
in any case (Table 3). After shelf-life conditions, fruit exhibited a significant
decrease with respect to the values presented just after storage
at 15 ◦ C. This decrease observed after astringency removal treatment
and subsequent shelf-life was due to the reduction <strong>of</strong> soluble
tannins responsible for fruit astringency, because they are included
in TSS measurements and they are reduced with astringency removal
treatment (Arnal and Del Río 2003). <strong>Ozone</strong> treatment did not
affect this parameter in both harvests. Changes in the values <strong>of</strong> pH
were not observed during storage at 15 ◦ C and subsequent shelf-life
period, maintaining similar values than initial in the 2 experiments
(data not shown). <strong>Ozone</strong> has no effect on this parameter.
Perez and others (1999) reported that sugar and organic acids
concentrations in strawberry were lower in ozone treatments than
in control samples; however, these differences were not observed
Figure 2 --- <strong>Effect</strong> <strong>of</strong> ozone
treatment on electrolyte
leakage (%) during storage
at 15 ◦ C and after
subsequent shelf-life period
at 20 ◦ C (90% RH) <strong>of</strong> “Rojo
Brillante” persimmon. Fruit
were submitted to
astringency removal
treatment prior to transfer
to 20 ◦ C.
S: Sensory & Nutritive Qualities <strong>of</strong> Food

S: Sensory & Nutritive Qualities <strong>of</strong> Food
<strong>Ozone</strong> effect on persimmon . . .
H1
CI (1000a/Lb)
H2
CI (1000a/Lb)
35
30
25
20
15
10
5
0
35
30
25
20
15
10
5
0
a
a
ab
a
b
ab
b
bc
c
c
Control <strong>Ozone</strong>
c
10 d 20 d 30 d 10d 20d 30d
c
ab
abc
ab
ab
bc
10 d 20 d 30 d 10d 20d 30d
a
days at 15∫ 15°C C days at 15∫ 15°C+5 C+5 d. at 20∫ 20°C C
days at 15∫ 15°C C days at 15∫ 15°C+5 C+5 d. at 20∫ 20°C C
Figure 3 --- <strong>Effect</strong> <strong>of</strong> ozone treatment on external color (CI)
during storage at 15 ◦ C and after subsequent shelf-life period
at 20 ◦ C (90% RH) <strong>of</strong> “Rojo Brillante” persimmon harvested
at 2 different dates: Harvest 1 (H1) and Harvest 2
(H2). Fruit were submitted to astringency removal treatment
prior to transfer to 20 ◦ C. Means for storage at 15
◦ C or for shelf-life periods with the same letter are not
significantly different at 5% level (LSD test).
Table 2 --- <strong>Effect</strong> <strong>of</strong> ozone treatment on acetaldehyde production
(mg/100 mL) during storage at 15 ◦ C and after
subsequent shelf-life period at 20 ◦ C (90% RH) <strong>of</strong> “Rojo
Brillante” persimmon harvested at 2 different dates: Harvest
1 and Harvest 2. Fruit were submitted to astringency
removal treatment prior to transfer fruit to 20 ◦ C.
a
Acetaldehyde (mg/100 mL)
Days at 15 ◦C Daysat15◦C + 5dat20◦C 10 20 30 10 20 30
Harvest 1
Control 0.17b 0.21c 0.13a 3.89e 2.96b,c 1.97a <strong>Ozone</strong> 0.12a 0.16b 0.16b 3.44c,d 3.13c 2.57b Harvest 2
Control 0.21c 0.20b,c 0.14a 3.30c 2.46a 3.48c <strong>Ozone</strong> 0.24d 0.18b 0.15a 2.80b 2.76a,b 3.48c a–e Data followed by the same letter for storage at 15 ◦ C or for shelf-life periods
did not differ at 5% significance level.
by Nadas and others (2003). <strong>Ozone</strong> treatment affects neither TSS
in pear and apple nor titratable acidity in apple during prolonged
storage (Skog and Chu 2001).
In the present research, any visual damage in control or ozonetreated
fruit was found under assayed conditions.
Conclusion
<strong>Ozone</strong> treatment had a positive effect on fruit firmness, this effect
being dependent on harvest date; in fruit from 2nd harvest,
ozone maintained fruit with commercial firmness values up to 30 d
plus shelf-life. <strong>Ozone</strong> increased weight losses in treated fruits; this
fact could be explained by an increase in EL. <strong>Ozone</strong> had no important
effects on other studied parameters. Therefore, ozone could be
considered a tool to maintain quality <strong>of</strong> persimmon during storage.
b
b
c
c
c
Table 3 --- <strong>Effect</strong> <strong>of</strong> ozone treatment on total soluble solids
(TSS) content during storage at 15 ◦ C and after subsequent
shelf-life period at 20 ◦ C (90% RH) <strong>of</strong> “Rojo Brillante”
persimmon harvested at 2 different dates: Harvest
1 and Harvest 2. Fruit were submitted to astringency removal
treatment prior transferring fruit to 20 ◦ C.
TSS ( ◦ Brix)
Days at 15 ◦C Daysat15◦C + 5dat20◦C 10 20 30 10 20 30
Harvest 1
Control 16.9a,b 16.7a,b 16.2a 14.4a,b 14.1a 14.7b <strong>Ozone</strong> 16.4a 17.0b 16.8a,b 14.2a,b 13.8a 14.2a Harvest 2
Control 18.9a 19.1a 18.7a 17.0a 17.2a,b 17.7a,b <strong>Ozone</strong> 19.3a 18.9a 19.1a 17.2a 18.1b 18.4b a–b Data followed by the same letter for storage at 15 ◦ C or for shelf-life periods
did not differ at 5% significance level.
Acknowledgments
The authors wish to thank D.O. kaki Ribera del Xúquer, Fomesa-
Fruitech Co. (Valencia, Spain), and Grupo Interozone Europa
(Lleida, Spain) for their financial and technical support.
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