Effect of Malic Acid and Calcium Treatments on Quality Characteristics of Apple Fruits During Storage
Apple texture can deteriorate during cold storage, resulting in softness and mealiness. The purpose of this work was to estimate shelf-life and to study the behavior of Jonagold apples kept at 0-2°C in a normal atmosphere. The experiment was started in season 2010-2011 and fruit weight losses, fruit firmness, Total Soluble Solid (TSS), pH, Titratable acidity, Total soluble solids/titratable acidity ratio, Thiault index, Perlim index, Ethylene production, Peroxidase and Catalase enzyme activities were measured at 20, 40, 60, 80, 100, 120 and 150 th days of postharvest life. The fruits were immersed in distilled water, three malic acid concentrations (0, 100 and 150 mg-1) or at three calcium concentrations (0, 0.35 and 0.7% w/v). Results showed that fruit weight loss significantly decreased in malic acid+calcium treatments in comparison to control. Also, results showed that malic acid+calcium treatments increase fruit firmness, Catalase activity (CAT), Titratable acidity (TA) and Perlim index while decreasing of pH, Total soluble solids/Titratable acidity ratio and Peroxidase activity (POD) during cold storage at 0-2°C for 5 month (p≤0.05). The results showed that malic acid+calcium treatments application was influenced on ethylene in comparison to control. In general, this experiment showed that post-harvest malic acid+calcium treatments prevented fruit softening and decreased weight losses.
Received: February 17, 2011;
Accepted: May 06, 2011;
Published: July 09, 2011
Apple fruits mostly stored after harvest. The fruits quality determination
includes color, surface damage, acidity, sugars and soluble solids (Omaima
and Haggag, 2007). The shelf-life of apples is affected by a number of factors,
such as growing, harvesting operations or storage conditions (Soliva-Fortuny
et al., 2002). With present day technology it is possible to store
apples in excellent physical condition well into the year following their harvest
but fruit quality depends strongly on temperature and humidity during storage
(Varela et al., 2005). Apple is a climacteric
fruit with a long post-harvest life in cool storage. Losses in fruit quality
are mostly due to its relatively high metabolic activity during storage (Fattahi
et al., 2010). Cool storage is widely used to reduce respiration
rate, ethylene production and extend the shelf-life of fruits. Calcium (Ca2+)
has been extensively reviewed as both an essential element and its potential
role in maintaining postharvest quality of fruit and vegetable crops. The role
of calcium in stabilizing cellular membranes and delaying senescence in horticultural
and agronomy crops is well known (Poovaiah et al.,
1988; Pervaiz et al., 2002; Hossain
et al., 2005; Abdi et al., 2006; Misra
and Gupta, 2006; Singh et al., 2006; Hosseini
and Thengane, 2007; Dzomeku et al., 2008;
Naeem et al., 2009; Al-Hamzawi,
2010). Pre- and postharvest application of calcium may delay senescence
in fruits with no detrimental effect on consumer acceptance (Lester
and Grusak, 1999). Exogenously applied calcium stabilizes the plant cell
wall and protects it from cell wall degrading enzymes (White
and Broadley, 2003). Faust (1989) asserted that Calcium
(Ca2+) in the apoplast exerts a binding effect in the complex of
polysaccharides and proteins comprising the cell wall and that cytoplasmic Calcium
(Ca2+) may regulate several enzyme activities. Postharvest calcium
dips can increase calcium content considerably compared to preharvest sprays,
without causing fruit injury, depending on salt type and calcium concentration.
Postharvest calcium application maintains cell turgor, membrane integrity, tissue
firmness and delays membrane lipid catabolism, extending storage life of fresh
fruits (Picchioni et al., 1998). Malic acid is
a well known organic acid that presumably can reduced the number of bacteria
in the solution and with decrease ACC-oxidase activity cause delay the onset
of hydrolysis of structural cell components, decrease ethylene production and
sensitivity (Kazemi et al., 2010). Previous study
had revealed that MA (malic acid) sprays during growth period increased chlorophyll
content of cut flowers while Ca Calcium spray caused extended post harvest vase
life (Darandeh et al., 2010). However, No evidence
was found for the hypothesis Malic acid application on the quality and storage
life of apple fruit during storage. In the present study, we test the hypothesis
Malic acid, calcium or their combination application on the quality and storage
life of apple fruit during storage.
MATERIALS AND METHODS
The experiment was started in season 2010-2011 and Fruit Weight Losses, Fruit Firmness, Total soluble solids, Titratable acidity, Total soluble solids/Titratable acidity, pH, Ethylene production, Thiault index, Perlim index, Peroxidase and Catalase enzyme activities were measured at 5 month of postharvest life. Malus domestica var. Jonagold was harvested at commercial maturity stage from an experiment orchard at the apple Research Institute of Iran (Zanjan, Iran). Fruits were subsequently transferred to laboratory and sorted based on size and the absence of physical injuries or infections. Fruits were randomly divided into four groups, each group containing 150 fruits in four replicates and immersed into solution of (0.35 and 0.7% (w/v) Ca, three malic acid concentrations (0, 100 and 150 mg-1) and in distilled water as control for 10 min. Fruits were then dried for about 24 h and then stored at 0-2°C and 85-90% relative humidity for 5 months. After 20, 40, 60, 80, 100, 120 and 150th days storage, 20 fruits per treatment were taken from cool storage for fruit quality assessment.
Weight loss: Physiological weight loss was determined by using Tefera
et al. (2007) method, by periodical weighing of apples 20, 40, 60,
80,100,120 and 150 days after storage. The differential weight losses were calculated
for each interval and converted into percentage by dividing the change with
the initial weight recorded on each sampling interval. The cumulative Physiological
weight loss was expressed in percent with respect to different treatments. The
difference between the initial weight and successive weights gave the rate of
weight loss (as percentages).
Fruit firmness: Firmness was measured on two opposite peeled sides using a pressure meter (OSK, 10576) fitted with an 8 mm diameter flat tip. The firmness considered as an average peak force of 10 fruits and expressed as kg.
Total soluble solid: TSS in the juice was determined with a hand-refractometer (NC-1, Atago Co., Japan) at room temperature and expressed as a percentage.
Titratable acidity: TA was determined by titration an aliquot (20 mL) of the juice to pH 8.2 with 0.1N NaOH and the result was expressed as a percentage of malic acid.
TSS/TA ratio: The maturity index was evaluated as the TSS/TA ratio (i.e.,
ratio increasing with maturity) (Schirra et al.,
pH: pH of the juice were measured using a pH meter (Jenway, 3020).
Peroxidase activity (POD): Peroxidase activity was measured based on
the method of Chance and Maehly (1955).
Catalase activity (CAT): CAT activity was measured based on the method
of Aebi (1984). The activity was measured in a reaction
mixture (2.0 mL final volume) composed of 30% H2O2 in
50 mM NaKPi, pH 7.0 and 1.5 mL of enzyme extract. Samples without H2O2
were used as a blank. The decomposition of H2O2 was followed
spectrophotometrically by the decrease in A240. Enzyme activity was expressed
in units of activity (U) mg-1 protein.
Ethylene determination: Three fruits were enclosed in 3 L airtight jars for 1 h at 20°C. Ethylene measurements were performed by withdrawing 1 mL headspace gas sample from the jars with a syringe and injecting it into a Varian 3300 gas chromatograph, equipped with a stainless steel column filled with Porapak, length 100 cm, diameter 0.32 cm, at 50°C and a flame-ionisation detector at 120°C. The carrier gas was nitrogen at a flow rate of 20 mL min-1.
Statistical analysis: All data were analyzed for significant differences using analysis of variance (ANOVA) using the SAS (Statistical Analysis System) statistical package (SAS Institute, Cary, NC, USA). Data were then subjected to mean separation by the Least Significant Difference test (LSD) at p<0.05.
RESULTS AND DISCUSSION
Weight loss: Effect of Ca and MA on weight losses of stored fruits are
listed in Table 1. Results showed that Dipped fruits in Ca+MA
solution at different concentration prevented weight loss in comparison with
control (p = 0.05). Maximum weight loss occurred in control treatment while
lowest loss was recorded in 150 mg-1 MA+ 0.7% (w/v) Ca (Table
1). Calcium applications have known to be effective in terms of membrane
functionality and integrity maintenance which may be the reason for the lower
weight loss found in Calcium treated fruits. The above mentioned results are
agreed with those recorded by Mahajan and Dhatt (2004).
They reported that pear fruit treated with Ca proved to be most effective in
reducing weight loss compared to non treated fruit during a 75 days storage
period. The above mentioned results are agreed with those recorded by Ashour
(2000). He found that Dipped fruits in Ca solution at different concentration
reduce apple weight losses percentages.
Firmness: It is clear from the obtained data in Table 1 that dipping apple in 150 mg-1 MA+0.7% (w/v) Ca were effective in Firmness for 5 month more than the other treatments in during storage.
||Mean comparison of fruit weight loss, firmness, ethylene,
TSS, TA, POD, CAT, TSS/TA, pH, thiault and perlim indexes in different concentration
Ca+MA solution during 5 month storage at 0-2°C
|Means in each column followed by similar letters are not significantly
different at 5% level (LSD test). 1Total soluble solids, 2Titratable
acidity, 3Peroxidase activity, 4Catalase activity
The results indicate that maximum firmness was recorded in 150 mg-1
MA+0.7% (w/v) Ca as compared to control, while minimum firmness was recorded
in control during 5 month (p = 0.05). The retention of firmness in calcium treated
fruits might be due its accumulation in the cell walls leading to facilitation
in the cross linking of the pectic polymers which increases wall strength and
cell cohesion (White and Broadley, 2003). This result
was in agreement with the report of Benavides et al.
(2002) that suggested postharvest application of apple by Ca decreased softening
and kept firmness during storage also, Casero et al.
(2004) reported that Dipped fruits in Ca solution at different concentration
increased apple Firmness percentages.
Total soluble solids, pH, Titratable acidity and TSS/TA ratio: TSS and
pH were not influenced by the postharvest calcium dips and slight differences
existed. Also, the results indicate that maximum TA was observed in 150 mg-1
MA+0.7% (w/v) and lowest TA was recorded in control. Titratable acidity is directly
related to the concentration of organic acids present in the fruit, which are
an important parameter in maintaining the quality of fruits. This result was
in agreement with the report of Manganaris et al.
(2005) that suggested postharvest application of apple by Ca dips did not
effect of TA % in peaches during storage. The TSS/TA ratio increased with increasing
the storage duration. But dipped fruits in Ca+MA solution at different concentration
prevented increasing of TSS/TA ratio in comparison with control (p≤0.05).
POD and CAT enzyme activities: The results indicate that maximum POD
activity was recorded in control as compared to 0.35 and 0.7 % (w/v) Ca and
other treatment (Table 1). Previous studies Lamikanra
and Watson (2001) indicated the ascorbate dependency of Peroxidase (POD)
enzymes in a number of commonly fresh-cut processed fruits whose activities
appear to be related to the level of oxidative stress in cut fruit. Ranadive
and Haard (1972) identified a correlation between peroxidase activity and
lignification in cell walls of pear fruit and demonstrated differences in peroxidase
activity at different Ca concentrations. Ca2+ appears to be necessary
because it induces the cross-linking of polygalacturonan chains into a structure
that can be recognized by its isoperoxidase (Penel et
al., 1999). The results in Table 1 show that the storage
period has a significant effect on Catalase activity (CAT) of fruits (p≤0.05).
The results indicate that maximum Catalase activity (CAT) was observed in 150
mg-1 MA+0.7% (w/v) while the lowest Catalase activity (CAT) was recorded
in control. High Ca concentrations result in decreased flesh browning symptoms
which are directly associated with calcium content in fruits (Hewajulige
et al., 2003).
Ethylene: The results in Table 1 show that the storage
period has a significant effect on ethylene of fruits (p = 0.05). The results
indicate that maximum ethylene was observed in control treatment, while, the
lowest ethylene was recorded in 150 mg-1 MA+0.7% (w/v). Oxidative
membrane injury allows the mixing of the normally separated enzyme (PPO) and
oxidizable substrates (polyphenols), which lead to browning (Hodges,
2003). High calcium concentrations result in decreased ethylene production,
electrolyte leakage and flesh browning symptoms which are directly associated
with calcium content in fruits (Hewajulige et al.,
2003). This result was in agreement with the report of Mortazavi
et al. (2007) that suggested postharvest application of apple by
Ca decreased electrolyte leakage and increased the cell wall integrity and stability.
Also, This result was in agreement with the report of Kazemi
et al. (2010) that showed the treatment MA treatment improved flower
quality with reduced ACC-oxidase activity (ACO), ethylene production and microbial
population in vase solution of carnation cut flowers. Ethylene possesses an
important role in integrating developmental signals and responses to abiotic
stresses, like cold storage and it has been suggested that calcium delays the
onset of the ethylene climacteric period and climacteric peak (Ben-Arie
et al., 1995).
The effect of MA on senescence indices, which is reported here for the first time, could be promising. MA and Calcium dips retarded metabolism as indicated by the lower respiration rates of MA+calcium treated samples. MA+calcium dips improved the firmness of apple. MA+Calcium concentration of treated samples was significantly greater (p≤0.05) than the control. Further studies are necessary to determine the sensory profile and the microbiological stability.
Aebi, H., 1984.
Catalase in vitro
. In: Methods in Enzymology, Academic Press, Cambridge, Massachusetts, ISBN: 9780121820053, pp: 121-126CrossRef | Direct Link |
Abdi, G., M. Khosh-Khui and S. Eshghi, 2006.
Effects of natural zeolite on growth and flowering of strawberry (Fragaria
Duch.). Int. J. Agric. Res., 1: 384-389.CrossRef | Direct Link |
Ashour, N.N., 2000.
Effect of environmental factors, calcium and potassium fertilization on yield and quality of apple. Ph.D. Thesis, Fac. Agric. Mansoura Univ, Egypt.
Ben-Arie, R., I. Mignani, L.C. Greve, M. Huysamer and J.M. Labavitch, 1995.
Regulation of the ripening of tomato pericarp discs by GA3
and divalent cations. Physiol. Plant., 93: 99-107.CrossRef |
Benavides, A., I. Recasens, T. Casero, Y. Soria and J. Puy, 2002.
Multivariate analysis of quality and mineral parameters on golden smoothee apples treated before harvest with calcium and stored in controlled atmosphere. Food Sci. Technol. Int., 8: 139-146.CrossRef | Direct Link |
Darandeh, N., E. Hadavi and M. Shoor, 2010.
Preharvest citric acid application extended post-harvest vase life of Lilium
L. Proccedings of 28th International Horticultural Congress, Lisbon Portugal. http://profdoc.um.ac.ir/paper-abstract-1017058.html.
Dzomeku, B.M., R.K. Bam, E. Adu-Kwarteng, A.A. Ankomah and S.K. Darkey, 2008.
Comparative study on the agronomic, nutritional values and consumer acceptability of fhia-21 (tetraploid hybrid) and apem (triploid french plantain) in Ghana. Am. J. Food Technol., 3: 200-206.CrossRef | Direct Link |
Casero, T., A. Benavide, J. Puy and I. Recasens, 2004.
Relationships between leaf and fruit nutrients an fruit quality attributes in Golden Smotthee apples using multivariate regression techniques. J. Plant Nutr., 27: 313-324.
Chance, B. and A.C. Maehly, 1955.
Assay of catalases and peroxidases: Catalase: 2H2
O + O2
(1), Catalase and peroxidase: ROOH + AH2
O + ROH + A (2). Methods Enzymol., 2: 764-775.CrossRef | Direct Link |
Fattahi, J., R. Fifall and M. Babri, 2010.
Postharvest quality of kiwifruit (Actinidia deliciosa
cv. Hayward) affected by pre-storage application of salicylic acid. S. Western J. Hortic. Biol. Environ., 1: 175-186.
Faust, M., 1989.
Utilization of Major Nutrients by Fruit Trees. In: Physiology of Temperate Zone Fruit Trees, Faust, M. (Ed.). John Wiley and Sons, New York, pp: 83-95
Hewajulige, I.G.N., R.S.W. Wijeratnam, R.L.C. Wijesundera and M. Abeysekere, 2003.
Fruit calcium concentration and chilling injury during low temperature storage of pineapple. J. Sci. Food Agric., 83: 1451-1454.CrossRef | Direct Link |
Hodges, D.M., 2003.
Postharvest Oxidative Stress in Horticultural Crops. Foods Products Press/The Howerth Press, Binghampton, New York
Hossain, A.K.M.Z., H. Koyama and T. Hara, 2005.
Sugar compositions and molecular mass distributions of hemicellulosic polysaccharides in wheat plants under aluminum stress at higher level of calcium supply. Asian J. Plant Sci., 4: 11-16.CrossRef | Direct Link |
Hosseini, G. and R.J. Thengane, 2007.
Salinity tolerance in cotton (Gossypium hirsutum
L.) genotypes. Int. J. Bot., 3: 48-55.CrossRef | Direct Link |
Kazemi, M., E. Hadavi and J. Hekmati, 2010.
The effect of malic acid on the bacteria populations of cut flowers of carnations vase solution. World Applied Sci. J., 10: 737-740.
Lamikanra, O. and M.A. Watson, 2001.
Effects of ascorbic acid on peroxidase and polyphenoloxidase activities in fresh-cut cantaloupe melon. J. Food Sci., 66: 1283-1286.CrossRef |
Lester, G.E. and M.A. Grusak, 1999.
Postharvest application of calcium and magnesium to honeydew and netted muskmelons: Effects on tissue ion concentrations, quality and senescence. J. Am. Soc. Hort. Sci., 124: 545-552.Direct Link |
Mahajan, B.V.C. and A.S. Dhatt, 2004.
Studies on postharvest calcium chloride application on storage behaviour and quality of Asian pear during cold storage. Int. J. Food Agric. Environ., 2: 157-159.Direct Link |
Al-Hamzawi, M.K.A., 2010.
Effect of calcium nitrate, potassium nitrate and anfaton on growth and storability of plastic houses cucumber (Cucumis sativus
L. cv. Al-Hytham). Am. J. Plant Physiol., 5: 278-290.CrossRef | Direct Link |
Manganaris, G.A., M. Vasilakakis, I. Mignani, G. Diamantidis and K. Tzavella-Klonari, 2005.
The effect of preharvest calcium sprays on quality attributes, physicochemical aspects of cell wall components and susceptibility to brown rot of peach fruits (Prunus persica
L. cv. Andross). Sci. Hortic., 107: 43-50.CrossRef |
Misra, N. and A.K. Gupta, 2006.
Interactive effects of sodium and calcium on proline metabolism in salt tolerant green gram cultivar. Am. J. Plant Physiol., 1: 1-12.CrossRef | Direct Link |
Mortazavi, N., R. Naderi, A. Khalighi, M. Babalar and H. Allizadeh, 2007.
The effect of cytokinin and calcium on cut flower quality in rose (Rosa hybrida
L.) cv. Illona. J. Food Agric. Environ., 5: 311-313.Direct Link |
Naeem, M., M. Masroor A. Khan and J.B. Morris, 2009.
Agrobotanical Attributes, nitrogen-fixation, enzyme activities and nutraceuticals of hyacinth bean (Lablab purpureus
L.): A bio-functional medicinal legume. Am. J. Plant Physiol., 4: 58-69.CrossRef | Direct Link |
Penel, C., P. van Cutsem and H. Greppin, 1999.
Interactions of a plant peroxidase with oligogalacturonides in the presence of calcium ions. Phytochemistry, 51: 193-198.CrossRef |
Pervaiz, Z., M. Afzal, S. Xi, Y. Xiaoe and L. Ancheng, 2002.
Physiological parameters of salt tolerance in wheat. Asian J. Plant Sci., 1: 478-481.CrossRef | Direct Link |
Picchioni, G.A., A.E. Watada, W.S. Conway, B.D. Whitaker and C.E. Sams, 1998.
Postharvest calcium infiltration delays membrane lipid catabolism in apple fruit. J. Agric. Food Chem., 46: 2452-2457.CrossRef | Direct Link |
Poovaiah, B.W., G.M. Glenn and A.S.N. Reddy, 1988.
Calcium and fruit softening: Physiology and biochemistry. Hortic. Rev., 10: 107-152.Direct Link |
Omaima, M. and H.E. Haggag, 2007.
Quality improvement and storability of apple cv. anna by pre-harvest applications of boric acid and calcium chloride. Res. J. Agric. Biol. Sci., 3: 176-183.
Ranadive, A.S. and N.F. Haard, 1972.
Peroxidase localization and lignin formation in developing pear fruit. J. Food Sci., 37: 381-383.
Schirra, M., M. Mulas, A. Fadda and E. Cauli, 2004.
Cold quarantine responses of blood oranges to postharvest hot water and hot air treatments. Postharvest Biol. Technol., 31: 191-200.CrossRef |
Singh, V.K., D.K. Singh and S.M. Pathak, 2006.
Relationship of leaf and fruit transpiration rates to the incidence of softening of tissue in mango (Mangifera indica
L.) cultivars. Am. J. Plant Physiol., 1: 28-33.CrossRef | Direct Link |
Soliva-Fortuny, R.C., G. Oms-Oliu and O. Martin-Belloso, 2002.
Effects of ripeness stages on the storage atmosphere, color and textural properties of minimally processed apple slices. J. Food Sci., 67: 1958-1963.CrossRef |
Tefera, A., T. Seyoum and K. Woldetsadik, 2007.
Effect of disinfection, packaging and storage environment on the shelf life of mango. Biosyst. Eng., 96: 201-212.CrossRef | Direct Link |
Varela, P., A. Salvador and S. Fiszman, 2005.
Shelf-life estimation of Fuji apples: Sensory characteristics and consumer acceptability. Postharvest Biol. Technol., 38: 18-24.CrossRef |