International Journal of Agriculture and Crop Sciences.
Available online at www.ijagcs.com
IJACS/2013/5-4/355-361
ISSN 2227-670X ©2013 IJACS Journal
Effects of pre-cooling and chlorine on post-harvest
quality ofcut roses (Rosa hybrida L.)
A.RezaeiNejad1,* and F.Nazarian2
1. Department of Horticultural Sciences, Faculty of Agriculture, Lorestan University, PO Box 465, Korramabad,
Iran
2. Department of Horticultural Sciences, Faculty of Agriculture, Islamic Azad University, Karaj Branch, Karaj,
Iran
*
Corresponding author email:[email protected]
ABSTRACT: The aim of this study was to quantify the vase-life and water relations of cut roses in
response to pre-cooling and different concentrations of chlorine in the hydration solution. Cut
‘Valentine’ roses were stored dry at 4oC or 25oC for 24h immediately after harvest and then placed in
vase-solutions containing 0, 50, 100 or 150 µl l-1chlorine provided as sodium hypochlorite. The results
showed significantly higher water loss in cut flowers dry stored at 25°C in the first 24 h after harvest
compared with that in cut flowers dry stored at 4°C. Acombination of dry storage at 4oCand inclusion of
50µl l-1chlorine in the vase-solutionextended vase-life by 6.1 d over the control (25oC and 0 µl l1
chlorine). Vase-solutions containing chlorine showed reduced microbial counts compared with
control. Results suggest that vase-life improvements are achieved through (1) a reduction in water
loss from cut stems and leaves during pre-cooling and (2) a reduction in bacterial populations present
in the vase-solution. This was associated with an increase in water uptake and relative fresh weight of
cut stems without any toxicity to leaves. The results also revealed that only two of eight rose cultivars
showed toxicity to leaves in response to 50 µl l-1chlorine, suggesting that toxicity in response to
effective chlorine concentration is cultivar dependent.
Keywords:Bacterial proliferation, Vase-life, Water relation.
INTRODUCTION
The vase-life of cut rose flowers is affected by various factors such as pre-harvest growing conditions and
post-harvest handling, storage and transportation
.All these factors may
result in a negative water balance (water loss >water uptake) causing wilting and bent neck. Physiologically, a
negative water balance could be due to: (1) poor water uptake as a result ofvascular occlusion, which inhibits
water supply to the flowers
!
"#$ %&
'
(
) *
and/or (2) a high water loss via transpiration trough stomata (Torreand Fjeld,
RezaeiNejad and van Meeteren, 2005).
The majority of fresh cut flowers sold in Iran are produced in the northern parts ofthe country. It generally
takes 24 h or longer, to commercially distribute these flowers to retailers throughout the country via truck
transport without any temperature control. It has been shown that commercial cut flowers are often exposed to
damaging high temperatures (Maxieet al. 1973; Thompson and Reid, 1994). Poor temperature management
during transport of cut flowers is largely the result of inadequate pre-cooling and transport under nonrefrigerated conditions. Leonard et al. (2011) showed that temperatures during the 24-hour transport system
increased steadily and temperatures were at or above 10 oC for 18 h, with half of that time above 15 oC for all
shipments. There are some reports on negative effects of improper storage temperatures on vase-life of a
range of cut flowers (Cevallos and Reid, 2000;Cevallos and Reid, 2001). Maxieet al. (1973) exposed ‘Improved
White Sim’ carnation for various lengths of time to typical packing temperatures and found that holdingflowers
for 24 h at 20 oC reduced vase life by 25% compared with holding at 0 oC. By contrast, a short retail simulation
display of 2 d found no difference in vase life of gerbera when kept at 2, 6, 10, and 21 oC (Nell et al., 2009). It
has been reported that reduction of cut flower vase-life during storage is highly correlated with respiration at the
storage temperature (+ !
*
Celikel and Reid, 2002). However, it is not clear to what extent
high temperatures after harvest during 24h dry storage of cut flowers correlate with changes in water relation
characteristics.
Previous studies have also shown that blockage of xylem vessels by micro-organisms that accumulate in
the vase-solution, or in the vessels themselves, is a major cause of deterioration in cut flowers (Zagory and
Intl J Agri Crop Sci. Vol., 5 (4), 355-361, 2013
( ,
%) #
#
& &* ! - -transpiration disrupts the water relations of cut
flowers (Knee, 2000) and consequently reduces vase-life (Halevy, 1976). Therefore, application of antimicrobial
compounds in the water may prolong the vase-life of cut flowers (Marousky,1976; van Doorn, 1997;Jowkar,
(
). A number of these compounds, however, are toxic to flowers, dangerous to human
health and/or pollute environment (Damunpola and Joyce, 2006).Sodium hypochloriteas a source of chlorine is
a cheap, environmentfriendly agent that can be used in the vase-solutions. Though it is widely used by
researchers (Cellavos and Reid, 2001),a detailed study has not been carried out on the effects of this agent on
flower vase-life. van Doorn et al. (1990) reported that when the concentration of the antimicrobial compound
containing chlorine was high enough to reduce the number of bacteria in stems to below the detection limit, the
roses (cv. Sonia) showed severe leaf chlorosis and leaf abscission. However the effect on other rose cultivars
is unknown.
In the present study, we aimed to elucidate the effects of chlorine on the vase and leaf life of cut rose
'Valentine'. Moreover, we sought to examine the effects of high temperature on vase-life and water relations of
cut rose flowers during 24 h of dry storage after harvest, simulating conditions before and during
transportationin Iran.
MATERIAL AND METHODS
Plant material and experimental conditions
Cut rose flowers (RosahybridaL.‘Valentine’) were obtained from a local commercial grower inIran. Flower
stems were harvested at normal harvest maturity (sepal starting to reflex) early in the morning, wrapped in
newspaper and transported to the laboratory of Lorestan University (a distance of 5 km). The weight of the cut
flowers was recorded and half of the cut flowers were then placed in a cold room at 4 ± 1 oC(Treatment 2) and
the other half were placed in the test room at 25 ± 1oC(Treatment 1)for 24 h. All cut flowers were wrapped in
newspaper and covered with black plastic bagsduring storage. After storage, flower stems were weighed again
to calculate the water loss during the first 24 h. The stemswere then re-cut under water to a stem-length of 45
cm and all leaves except for the upper three were removed.Single flower stems were kept in 500 ml conical
flasks, each containing 400 ml vase-solutionsof 0, 50, 100 or 150µl l-1chlorine using sodium hypochlorite (13%
active chlorine, AcrosOrganics N.V., NJ).Distilled water was used for vase-solution preparations. The mouths of
the flasks were covered withaluminum foil to prevent evaporative water loss. The experimental conditions were a
temperature of 25 ± 2oC, 60% relative humidity, and a 12-h photoperiod under 10 µmolm-2s-1 irradiance provided
by cool-white fluorescence lamps.
Measurements of vase-life, leaf-lifeand water relation parameters
The fresh weight of cut flowers and the amount of water uptake were measured daily. Longevity of flowers
and leaves was recorded as vase-life and leaf-life(in d), from the time the cut flowers were placed in conical
flasks (day-0). Flowers were considered to be at the end of theirvase-life when the whole flower wilted
(Pompodakis and Joyce, 2003 ,&%
).The leaf-life was considered to be at the end when two of
the three leaves on each stem wilted, dried or abscised.
Bacteria enumeration
At 2 d intervals, sampling from each vase-solution was carried out until day-6 of the experiment. After serial
dilutions, samples of vase-solution were plated onto nutrient agar and incubated for 48 h at 30°C. The number of
micro-organisms was counted by the standard plate counting method to determine the number of Colony Forming
Units ml-1 (CFU ml-1) (Jowkar, 2006).
Cultivar differences in response to sodium hypochlorite
To test the response of different rose cultivars to sodium hypochlorite, in a separate experiment, eight cut
rose cultivars, namely ‘Valentine’, ‘King Pride’, ‘Prima Donna’, ‘Black Baccara’, ‘Victory’, ‘Papagayo’, ‘Royal
Baccara’ and ‘Yellow Island’ were obtained from the same company as used above and transported to the
laboratory. Flower stems were placed in tap water and used for experiments within one hour after harvest.
Flower stems were re-cut under water to a stem-length of 45 cm and all leaves except for the upper three
leaves were removed. Single flower stems were kept in 500 ml conical flasks, each containing a 400 ml vasesolution of 50 µl l-1chlorine provided assodium hypochlorite. Distilled water was used for vase-solution
preparation. The abovementioned conditions were applied and leaf-lifewas recorded.
Data collection and statistical analysis
Storage of cut flowers at 4 °C and 25 °C during the first 24 h after harvest and inclusion of four
concentrations of chlorine using sodium hypochlorite (0, 50, 100 and 150 µl l-1) in the vase-solutions was
combined factorially based on a randomised complete block design (RCBD) with four replications. The
Intl J Agri Crop Sci. Vol., 5 (4), 355-361, 2013
experiment for cultivar differences was carried out according to a RCBD with five replications. Statistical
significance between mean values was assessed using analysis of variance (ANOVA) and Duncan's Multiple
Range Test at P < 0.05. MSTAT-C software (Michigan State University, East Lansing, MI, USA) and GraphPad
Prism 4 for Windows (GraphPad Software, San Diego, CA, USA) were used for all statistical analyses.
RESULTS AND DISCUSSION
Water loss during first 24 h
Significantly (P<0.01) higher water loss was found in cut flowers dry stored at 25°C in the first 24 h after
harvest compared with that in cut flowers dry stored at 4°C. Cut flowers stored at 25°C in the first 24 h after
harvestlost 12.1 ± 0.6% of their fresh weight compared with 7.0 ± 0.5% loss of fresh weight in cut flowers stored
at 4°C.
Vase-life and leaf-life
Pre-cooling after harvest and inclusion of sodium hypochlorite significantly (P<0.05) increasedthe vase-life
and leaf-lifeof cut roses 'Valentine' (Figure 1). The combination of pre-cooling and 50 µl l-1chlorine extended
vase-life and leaf-life by 6.1 and 6.3 d (56.3 and 60.9%), respectively, over the control (25°C and 0 µl l-1
chlorine).
Fresh weight and water uptake
There were significant differences (P<0.01) in daily changes in relative fresh weight (RFW) and water
uptake (WU) of cut rose 'Valentine' dry stored at 25 (T1) or 4oC(T2) for the first 24h after harvest and then
placed in vase-solutions containing 0, 50, 100 or 150 µl l-1chlorine (Figures 2 and 3).The combination of precooling and 50 µl l-1chlorine (T2C2) gave the highest RFW and WU, followed by cold storage plus 100 µl l1
chlorine (T2C3). Fast decreases in RFW and WU were found in warm storage plus 150 µl l-1chlorine (T1C4)
followed by the control treatment (T1C1).
Number of bacteria in vase-solutions
There was a significant difference (P < 0.01) in changes over time of the number of bacteria in vasesolutions containing 0, 50, 100 or 150 µl l-1chlorine (Figure4). Vase-solutions containing 0 µl l-1chlorine (T1C1
and T2C1) showed a very rapid microbial growth over time. Until day-2 no microbes were found in the 150 µl l1
chlorine treatments (T1C4 and T2C4). The lowest microbial counts were found in these treatments. Vasesolutions containing 50 and 100 µl l-1chlorine (T1C2, T2C2, T1C3 and T2C3) showed reduced microbial counts
compared with control treatments but higher than in 150 µl l-1chlorine treatments.
Cultivar responses to sodium hypochlorite
There was a significant difference (P<0.01) in the leaf-lives of the cultivars in response to sodium
hypochlorite. Inclusion of 50 µl l-1chlorine using sodium hypochlorite in the vase-solution resulted in severe
toxicity in two cultivars:‘Royal Baccara’ and ‘Yellow Island’ (Figure5). However, most cultivarsi.e. ‘Valentine’,
‘King Pride’and ‘Victory’showed high leaf-life when placed in solutions containing sodium hypochlorite.
Our results showed that high temperatures (25°C) after harvest during short-term(24 h) dry storage of cut
flowers (conditions simulating before and during transportation in Iran)not only caused higher water loss in that
period but also negatively affect water relation characteristics during subsequent vase life. It seems that dry
storage at high temperature caused a severe water deficit in stem and leaves which could not be repaired,
even afterre-cutting stems and placing in vase-solution. Higher water loss may lead to the development of air
emboli in the xylem whichcould not be removed by re-cuttingof a few centimeters of the basal part of the stem
(van Meeteren et al., 2005). The results showed that pre-cooling during 24 h dry storage of cut flowers, reduced
water deficit in stems and leaves, thus contributing to a longer vase-life of cut roses (Reid, 2001).Moreover,
another possibility to extend the vase-lifeis wet storage of cut flowers during shipment at high temperatures, as
shown by Cevallos and Reid (2001).
It has been reported that when the concentration of the antimicrobial compound containing chlorine was
high enough to reduce the number of bacteria in stems, the roses (‘Sonia’) showed severe leaf chlorosis and
leaf abscission (van Doorn et al., 1990). According to our results, the concentration of 50 µl l-1chlorine was
enough to extend longevity of cut rose ‘Valentine’without any toxicity to leaves.There are some reports
indicating that electrolyzed neutral water containing 15 to 50 µl l-1available chlorine reduced the microbial
population on fresh cut vegetables and in flower vases and stems of cut rose ‘Asami Red’ without causing
discoloration,chlorosis or leaf abscission ".&%
".&%
1). Ourexperiment with eight cut rose cultivars
revealed that only two cultivars, namely ‘Royal Baccara’ and ‘Yellow Island’, showed severe leafchlorosis and
leaf abscission in response to 50 µl l-1chlorine. These resultsindicate that toxicity in response to effective
concentration of chlorine is cultivar dependent.
Intl J Agri Crop Sci. Vol., 5 (4), 355-361, 2013
16
2 5 °C
4 °C
A
a
Vaselife(d)
ab
12
c
c
b
c
8
d
d
4
0
50
100
150
C h l o r i n e ( µ l l -1 )
a
Leaf life(d)
16
2 5 °C
4 °C
B
ab
bc
bc
ab
12
c
c
100
150
c
8
4
0
50
C h l o r i n e ( µ l l -1 )
Figure 1. Vase-life (Panel A) and leaf-life (Panel B) of Rosa hybrida'Valentine' cut flowers dry stored at 25 (T1) or 4oC (T2)
for the first 24 h after harvest and then placed in vase-solutions containing 0, 50, 100 or 150 µl l-1 chlorine using sodium
hypochlorite. Data are the means of four replicates ± standard errors. Bars indicate standard errors larger than symbols.
Different letters show significant difference among means (P<0.05).
140
(%of initial value)
Relative freshweight
T1C 1
T1C 2
T1C 3
T1C 4
A
130
120
110
100
90
80
70
60
0
1
2
3
4
5
6
7
8
9
10
11
Tim e (d)
140
(%ofinitial value)
R
elativefreshw
eight
130
T
T
T
T
B
120
110
C
C
2C
2C
2
1
2
2
3
4
100
90
80
70
60
0
1
2
3
4
5
6
7
8
9
10
11
T im e (d )
Figure 2. Daily changes in relative fresh weight of Rosa hybrida 'Valentine' cut flowers dry stored at 25 oC (T1) (Panel A) or
4oC (T2) (Panel B) for the first 24 h after harvest and then placed in vase-solutions containing 0, 50, 100 or 150 µl l-1
chlorine using sodium hypochlorite (C1, C2, C3 and C4, respectively). Data are the means of four replicates ± standard
errors. Bars indicate standard errors larger than symbols.
Intl J Agri Crop Sci. Vol., 5 (4), 355-361, 2013
-1
-1
Water uptake (ml d gFW )
0 .9
T1C 1
T1C 2
T1C 3
T1C 4
A
0 .8
0 .7
0 .6
0 .5
0 .4
0 .3
0 .2
0 .1
0
1
2
3
4
5
6
7
8
9
10
11
Tim e (d)
-1
-1
Water uptake(ml d gFW )
0 .9
T2C
T2C
T2C
T2C
B
0 .8
0 .7
0 .6
1
2
3
4
0 .5
0 .4
0 .3
0 .2
0 .1
0
1
2
3
4
5
6
7
8
9
10
11
Tim e (d)
10
7
10
6
10
5
10
4
10
3
10
2
10
1
10
0
T1C
T1C
T1C
T1C
T2C
T2C
T2C
T2C
1
Number of bacteria(CFUml- )
Figure 3. Daily changes in water uptake of Rosa hybrida 'Valentine' cut flowers dry stored at 25 oC (T1) (Panel A) or 4oC (T2)
(Panel B) for the first 24 h after harvest and then placed in vase-solutions containing 0, 50, 100 or 150 µl l-1 chlorine using
sodium hypochlorite (C1, C2, C3 and C4, respectively). Data are the means of four replicates ± standard errors. Bars indicate
standard errors larger than symbols.
10
1
2
3
4
1
2
3
4
-1
0
2
4
6
Tim e (d)
Figure 4. The number of bacteria that accumulated in vase-solution of Rosa hybrida'Valentine' cut flowers dry stored at 25
(T1) or 4oC (T2) for the first 24 h after harvest and then placed in vase-solutions containing 0, 50, 100 or 150 µl l-1 chlorine
using sodium hypochlorite (C1, C2, C3 and C4, respectively). Data are the means of four replicates ± standard errors. Bars
indicate standard errors larger than symbols.
Intl J Agri Crop Sci. Vol., 5 (4), 355-361, 2013
18
16
14
Leaf life (d)
12
10
8
6
4
2
Pa
pa
ga
yo
Ro
ya
lB
ac
ca
ra
Y
el
lo
w
Is
la
nd
ic
to
ry
V
Ba
cc
ar
a
on
na
Bl
ac
k
aD
Pr
id
e
Pr
im
in
g
K
V
al
en
tin
e
0
Cultivars
Figure 5. Leaf-life of eight cut rose cultivars placed in vase-solutions containing 50 µl l-1 chlorine using sodium hypochlorite.
Data are the means of five replicates ± standard errors.
In conclusion, a combination of dry storage at 4oC and inclusion of 50 µl l-1chlorine in the vase-solution
extended vase-life by 6.1 d over that of the control (25oC and 0 µl l-1chlorine). The results suggest that vase-life
improvements are achieved through (1) a reduction in water loss from cut stems and leaves during pre-cooling;
and (2) a reduction in bacterial populations present in the vase-solution due to chlorine. This was associated
with increased water uptake and relative fresh weight of cut stems without any toxicity to leaves. The results
also revealed that only two of eight rose cultivars showed toxicity in response to 50 µl l-1chlorine, suggesting
that toxicity in response to effective concentration of chlorine is cultivar dependent.
ACKNOWLEDGEMENT
The authors wish to thank KobraSepahvand for her technical assistance.
REFERENCES
Celikel FG, Reid MS. 2002. Storage temperature affects the quality of cut flowers from the Asteraceae. HortSci 37: 148-150
Cevallos JC, Reid MS. 2000. Effects of temperature on the respiration and vase-life of Narcissus flowers.ActaHort 517: 335-341
Cevallos JC, Reid MS. 2001.Effect of dry and wet storage at different temperatures on the vase-life of cut flowers.HortTech 11:199-202
DamunupolaJW, Joyce DC. 2006. When is a vase-solution biocide not, or not only, antimicrobial? J JpnSocHorticSci77: 1-18
De Stigter HCM. 1980. Water balance of cut and intact Sonia rose plants. ZPflanzenphysiol 99: 131–140
HalevyAH. 1976. Treatments to improve water balance of cut flowers. ActaHort64:223–230
Ichimura K, Kojima K, Goto R. 1999. Effects of temperature, 8-hydroxyquinoline sulphate and sucrose on the vase-life of cut rose flowers.
Postharvest BiolTechnol 15:33–40
Izumi H. 1999.Electrolyzed water as a disinfectant for fresh-cut vegetables.J Food Sci 64:536-539
Izumi H. 2001. Effects of electrolyzed neutral water on the bacterial populations in a flower vase and in stems of cut roses. J
JpnSocHorticSci70:599-601
Jin J, Shan N, Ma N, Bai J,Gao J. 2006. Regulation of ascorbate peroxidase at the transcript level is involved in tolerance to postharvest
water deficit stress in the cut rose (Rosa hybridaL.) cv. Samantha. Postharvest BiolTechnol 40:236–243
Jowkar MM. 2006.Water relations and microbial proliferation in vase-solutions of Narcissus tazettaL. cv. ‘Shahla-e-Shiraz’ as affected by
biocide compounds.J HorticSci Biotech 81:656–660
Knee M. 2000.Selection of biocides for use in floral preservatives.Postharvest BiolTechnol18: 227–234
Kumar N, Srivastava GC, Dixit K. 2008. Effect of ethanol plus sucrose on the vase-life of cut rose (Rosa hybridaL.). J HorticSci
Biotech83:749–754
Leonard RT, Alexander AM, Nell TA. 2011. Postharvest Performance of Selected Colombian Cut Flowers after Three Transport Systems to
the United States. HortTech 21:435-442
Marousky FJ. 1976. Control of bacteria in vase water and quality of cut flowers as influenced by sodium dichloroisocyanurate, 1, 3-dichloro5, 5-dimethylhydantoin and sucrose.United States Department of Agriculture, Agricultural Research Service S-115.
Maxie EC, Farnham DS, Mitchell FG, Sommer NF, Parsons RA, Snyder RG, Rae HL. 1973. Temperature and ethylene effects on cut
flowers of carnations (Dianthuscaryophyllus). J Am SocHorticSci 98:568-572
Mayak S, Halevy AH, Sagie S, Bar-Yoseph A, Bravdo B.1974. The water balance of cut rose flowers. PhysiolPlantarum31: 15–22
Nell TA, Leonard RT, Alexander AM. 2009. The effects of retail display conditions on postharvest performance of cut Gerbera jamesonii.
ActaHort 847:51–58
Pompodakis NE, Joyce DC. 2003. ABA analogue effects on vase-life and leaf crisping of cut ‘Baccara’ roses. Aust J Agr Res 43:425–428
Intl J Agri Crop Sci. Vol., 5 (4), 355-361, 2013
Reid M. 2001. Advances in shipping and handling of ornamentals. ActaHort 543: 277-284
Rezaei Nejad A,van Meeteren U. 2005.Stomatal response characteristics of Tradescantiavirginiana grown at high relative air
humidity.PhysiolPlantarum125: 324-332
Robinson T, Graham M, Dixon A, Zheng Y. 2009. Aqueous ozone can extend vase-life in cut rose.J HorticSci Biotech 84: 97–101
Solgi M, Kafi M, Taghavi TS, Naderi R.2009.Essential oils and silver nanoparticles (SNP) as novel agents to extend vase-life of gerbera
(Gerbera jamesonii cv. ‘Dune’) flowers.Postharvest BiolTechnol 53:155-158
Thompson JF, Reid MS. 1994. Temperature management during transportation of California cut flowers. Final Report of California Cut
flower Commission.
Torre S, Fjeld T. 2001.Water loss and postharvest characteristics of cut roses grown at high or moderate relative air
humidity.SciHortic89:217-26
vanDoorn WG, De Witte Y, Perik RRJ.1990. Effect of antimicrobial compounds on the number of bacteria in stems of cut rose flowers.
JApplBacteriol68: 117–122
vanDoorn WG. 1997. Water relations of cut flowers. Hort Rev18: 1–85
vanMeeteren U, van Gelder A, van Ieperen W. 2005.Effect of growth conditions on post-harvest rehydration ability of cut chrysanthemum
flowers.ActaHort 669: 287-296
Zagory D, Reid MS. 1986.Role of vase-solution microorganisms in the life of cut flowers.J Am SocHorticSci111: 154–158