Pitted Fruit Research Articles

Impact of postharvest dips with abscisic acid, prohexadione, calcium, or water on bitter pit incidence and apple physiology

Bitter pit causes significant losses to apple producers, packers, and retailers each year. While bitter pit is often associated with calcium deficiency, this postharvest disorder is still not fully understood. Some studies have demonstrated positive effects of preharvest sprays with prohexadione and abscisic acid. To evaluate the effects of these phytohormones as postharvest dips, ‘Granny Smith’ apples were dipped after harvest in prohexadione-Ca, abscisic acid, or CaCl2. Undipped and water dipped fruit were included as controls. Bitter pit incidence and severity were evaluated on fruit stored at 0 °C and >90 % RH for 68–75 d. Postharvest prohexadione-Ca and abscisic acid treatments did not reduce bitter pit incidence over two years of testing. However, there was an increase in bitter pit incidence in control fruit dipped in water and surfactant compared to undipped control fruit in the first year of testing. This increase (7.5–14 %) was observed again in water dipped control fruit in each of the following 2 years of experimentation. Inclusion of 1 % calcium chloride in the dip solution eliminated this increase in bitter pit incidence. Applying calcium with a surfactant increased the apoplastic calcium concentration and reduced bitter pit development compared to water dipped fruit. Results indicate that the increase in bitter pit induced by water dips may be due to removal of residual calcium on the fruit’s surface from preharvest calcium treatments. Ethylene production was higher in bitter pit fruit compared to healthy fruit. Reduced and total ascorbate were decreased in pitted fruit compared to healthy calcium dipped fruit. Dichlorofluoresceine diacetate fluorescence was higher in tissue adjacent to pits compared to healthy tissue from healthy fruit, indicating accumulation of reactive oxygen species. However, no consistent trend was observed in antioxidant enzyme activity. These results indicate that bitter pit shares some, but not all, of the oxidative metabolic trends observed in other fruit calcium deficiency disorders.

Characterisation of Position‐Dependant Ripening Dynamics of Nectarines Using Near‐Infrared Spectroscopy and ASCA

ABSTRACTNectarines, a popular pit fruit closely related to peaches, are renowned for their nutritional value and associated health benefits. However, challenges arise in maintaining optimal organoleptic properties during harvest and handling, eventually leading to production waste and heterogeneous quality in the fruit that arrives to the consumer. This study investigates the impact of nectarine position on trees during the whole ripening process using non‐destructive near‐infrared (NIR) spectroscopy. Nectarines exposed to more sunlight mature faster and this influences sugar content and acidity, emphasising the significance of considering height, prominence and orientation in ripening dynamics of the fruit. Different data unfolding strategies were compared, using ANOVA‐Simultaneous Component Analysis (ASCA) to reveal the significance of in‐tree position factors at different ripening stages, and observing high significance at harvest. This underscores the necessity for growers and handlers to consider these factors for reducing waste. NIR spectroscopy, with adequate data analysis, is a valuable tool for the holistic analysis of fruit ripening, providing crucial insights for maintaining optimal fruit organoleptic properties from harvest to consumer.

The Effect of Birch Pollen Immunotherapy on Apple and rMal d 1 Challenges in Adults with Apple Allergy

Background: A proportion of patients allergic to birch pollen are also allergic to pit fruit. The objective of this study was to investigate the effect of immunotherapy with birch pollen on birch-pollen-related apple allergy. Method: Patients with birch pollen immunotherapy underwent a skin-prick test with birch pollen, apple and rMal d 1, global assessments and nasal challenges with birch pollen, open food challenge with apple and a double-blind, placebo-controlled test with rMal d 1 at the start of and during the immunotherapy. Measurements of specific IgE in response to Bet v 1 and rMal d 1 and IgG4 in response to Bet v 1 and rMal d 1 took place. Results: Six of eight patients demonstrated an improvement of nasal challenge test results and all patients improved on global assessment during the immunotherapy. The median oral dose of apple required to elicit a reaction increased but was not statistically significant. The patients showed a decrease in skin-prick test values in response to birch pollen (1.05 to 0.36), apple (0.78 to 0.25) and rMal d 1 (0.51 to 0.10) with p-values of 0.04, 0.03 and 0.06, respectively and a decrease of specific IgE in response to Bet v 1 (10.66 kU/L to 5.19 kU/L) and rMal d 1 (0.99 to 0.61 kU/L) with p-values of 0.01 and 0.05, respectively. Only the median specific IgG4 value to Bet v 1 increased from 0.05 to 1.85 mg/L (p-value of 0.02) and not to IgG4 rMal d 1 (0.07 to 0.08 kU/L). Conclusion: The beneficial effects of immunotherapy for birch pollen were accompanied by a limited effect on apple allergy.

성목기 ‘감홍’/M.9 사과나무의 착과수준이 고두증상 및 과실품질에 미치는 영향

성목기 ‘감홍’/M.9 사과나무의 착과수준이 고두증상 및 과실품질에 미치는 영향

Effect of Sulfur and Potassium Application under Salinity Stress on Productivity and Fruit Quality of Swelling Peach Cultivar

This study was conducted during two successive seasons (2016 and 2017) at El-Sheikh Zwayed, North Sinai Governorate, Egypt. The aim of this research was to investigate the effect of three anti-salt stress materials (Nile fertile, elemental sulfur and potassium sulfate) on Swelling peach cultivar under irrigation water saline stress. In this experiment Nile fertile was applied at (0, 500 and 1000 gm/ tree), elemental sulfur (95% S) as soil application at three levels (0, 500 and 750 g / tree) and potassium sulphate as foliar application on three concentrations (0, 2, and 4 g/l.) and combination of them. The obtained results revealed that all anti-salt stress substances have a positive effect on vegetative growth parameters, fruit set, and yield/tree. The highest significant value of the final fruit set, yield/tree was obtained with Nile fertile treatment followed by elemental sulfur and potassium sulphate in both seasons of this study. On the other hand, there was no significant differences between the three materials on fruit length, fruit diameter, flesh and pit fruit weight, total soluble solids, total acidity and leaf content of N%, P% and k% during both seasons of the study and leaf content of Fe (ppm) in the 1st season. While the interaction effect between the three investigated factors shows clearly significant effects in improving vegetative growth, final fruit set no. of fruits /tree, fruit weight, crop yield, fruit physical and chemical parameters. Thus, the combination between Nile fertile 1000g/tree + elemental sulfur 750g/tree + potassium sulphate at (4g/L) gave the best results on shoot length, leaf area, initial, final fruit set %, fruit length and diameter, flesh weight (g), pit weight (g), maximum yield/tree, TSS, lowest acidity%. In addition to N, P, K, Fe, Zn and Mn in leaves in both seasons. On the other hand, untreated trees recorded the lowest values in previous studied parameters in this respect.

Vacuum Drying for Extending Litchi Shelf-Life: Vitamin C, Total Phenolics, Texture and Shelf-Life Assessment.

In an attempt to obtain shelf-stable litchi fruit with preserved nutritional quality and good sensory features, quarters of peeled and pitted fruits were vacuum dried at 50, 60 and 70°C at a constant pressure of 8.0kPa. The product was assessed for its vitamin C, total phenolics and texture (hardness). In addition, the product with the best texture was assessed for its shelf-life by means of accelerated testing. Results suggest that vacuum dried litchi retained almost 70% of the vitamin C and total phenolics when compared to frozen fruits (control). Vitamin C and phenolic compounds content significantly decreased with drying, while no difference was found between different drying temperatures. Hardness increased with drying temperature. The sample dried at 70°C presented crispness, which is a desired quality feature in dried fruit products. This sample was subjected to shelf-life evaluation, whose result suggests a shelf-life of eightmonths at 23°C. Total color change (CIE ΔE00) was the expiry criterion. Vacuum drying was a suitable technique for producing shelf-stable litchi fruit with good texture while preserving its desirable original nutrients. Consumption of vacuum dried litchi may be beneficial to health due to its remarkable content of phenolic compounds and vitamin C.

Identification of bitter pit protein markers in Malus domestica using differential in-gel electrophoresis (DIGE) and LC–MS/MS

Bitter pit is a physiological disorder that occurs in apple, pear and quince and has long been associated with calcium uptake or lack thereof. In the present study, pooled biological Malus domestica proteins were collected from healthy and naturally occurring bitter pit fruit. Protein samples (bitter pit and healthy) were analyzed with differential in-gel electrophoresis (DIGE) and SameSpots software was used to compare gel spots by intensity. Identified spots (p<0.05) were spot picked and trypsin digested. Peptides were separated by liquid chromatography (LC) and submitted to LTQ-Orbitrap mass spectrometer to infer protein identification. A total of 200±5 protein spots were detected, 41 spots classified as having p<0.05 and were successfully identified by their peptide sequence listed in an online M. domestica database. Thirteen spots were identified as having p<0.05 and a minimum 2-fold change. Several pathogenesis-related (PR) proteins belonging to three PR families, PR-5 (10.3-fold), PR-8/chitinase (35.5-fold) and PR-10 (6.8-fold) were upregulated in bitter pit sampled tissues. Three proteins involved in several metabolic processes including ethylene biosynthesis (3.3-fold), glycosyltransferase reactions in metabolism (2.3-fold) and metal biding (11.1-fold) were found to be overly expressed in healthy sampled tissues. This research provides a significant advance in the knowledge of protein expression alterations occurring in bitter pit in comparison with sound tissues.

Efeitos da rega e do regime hídrico em olival super intensivo no Alentejo

This study characterizes the effects of a typical farmer irrigation management and one of deficit irrigation imposed to a hedgerow olive orchard (1, 35 x 3,75 m) in the region of Evora, Southern Portugal on a Eutric Cambisol soil (FAO, 1998). The two irrigation regimes and their effects on tree transpiration, soil water content and stem and leaves water potential, a physiological parameter indicator of plant water stress, are analysed. With deficit irrigation DI imposed after pit fruit hardening, it was observed a progressive reduction on the values of tree transpiration, stem and leaf water potential and soil water content predominantly towards the end August and September. For the normal irrigation treatment (FI) transpiration values slightly declined for the months of July and August, although less than the ones for treatment DI. Results obtained with the “big leaf” model of Penman-Monteith (Jones, 1992) and bulk canopy conductance model of Orgaz et al. (2007) indicate that once validated they can be used to predict the transpiration of non-stressed hedgerow olive orchards in Alentejo. Predicting transpiration with the remotely sensed vegetation index NDVI proved difficult for lack of a strong correlation between the two parameters.

One Dimensional Linescan x-ray Detection of Pits in Fresh Cherries

The presence of pits in processed cherries is a concern for both processors and consumers, in many cases causing injury and potential lawsuits. While machines used for pitting cherries are extremely efficient, if one or more plungers in a pitting head become misaligned, a large number of pits may pass before corrective action is taken. While x-ray imaging has the potential to detect pits, traditional commercially available equipment is expensive and bulky, and implementation on the processing line is cumbersome. An x-ray inspection system using an array of photodiode based x-ray detectors in a linescan configuration whose outputs are combined to produce a one dimensional signal would be simpler, faster, and more economical. The data collection process is then reduced from a two dimensional image to a much simpler one dimensional signal, resulting in faster and simpler processing and classification. An algorithm designed to differentiate unpitted from pitted cherries for such a system yielded recognitiothe unpitted cherries, with a total error rate of 3.5%. When the a of pitted fruit, 100% of pitted cherries were detected with a orientation is controlled after pitting, total error is reduced to 1%.

Cellular approach to understand bitter pit development in apple fruit

Bitter pit (BP), a Ca 2+ deficiency disorder of apple fruit ( Malus domestica), is a complex process that involves not only the total input of Ca 2+ into the fruit, but also a proper Ca 2+ homeostasis at the cellular level. The objective of this study was to test the hypothesis that Ca 2+ accumulation into storage organelles and binding to the cell wall is associated with BP development in apple fruit. The experiment was carried out on ‘Granny Smith’ apples stored at 0 °C for 60 d. After storage, fruit were segregated into two lots for analysis, apples with the water-soaked initial visual symptoms of BP and those not showing this symptom. Cytochemical and ultrastructural observations showed an accumulation of Ca 2+ in the vacuole of individual outer cortical cells of pitted fruit. We also observed an increase in the expression of genes encoding four pectin methylesterases, a greater degree of pectin deesterification and therefore more Ca 2+ binding sites in the cell wall, and a higher fraction of the total cortical tissue Ca 2+ content that was bound to the cell wall in pitted fruit compared with non-pitted fruit. Cells of the outer cortical tissue of pitted fruit consistently had higher membrane permeability than outer cortical cells of non-pitted fruit. The results provide evidence that Ca 2+ accumulation into storage organelles and Ca 2+ binding to the cell wall represent important contributors to BP development in apple fruit.

Low light and low leaf calcium and boron concentrations are associated with fruit pitting, a new disorder in mango (Mangifera indica L.) in India

SummaryA study was conducted to investigate factors associated with fruit pitting, a new physiological disorder in mango (Mangifera indica L.). The incidence of pitting, along with intercepted light (photosynthetic photon flux density; PPFD) and net CO2 assimilation (A) were measured over 2 years at the top of the tree, and 1.0 m and 0.5 m above the crotch (i.e., the main branch-point) in the SE, NE, NW, and SW directions of the tree canopy in the mango cultivars ‘Alphonso’, ‘Amrapali’, ‘Bangalora’, ‘Bombay Green’, ‘Chausa’, ‘Dashehari’, ‘Langra’, ‘Neelum’, ‘Sensation’ and ‘Tommy Atkins’. Leaf nutrient concentrations were also determined in branches bearing normal and pitted fruit. The incidence of pitting was lowest (6.3%) in fruit at the top of the tree, followed by fruit 1.0 m (13.6%) or 0.5 m (20.0%) above the crotch. Similarly, the incidence of pitting was lowest (4.9%) in fruit from the SE of the canopy, followed by those from the NE (8.3%), SW(13.6%), and NW (20.0%). The incidence of pitting in different parts of the canopy reflected the PPFD and A values (A = 0.03 PPFD – 5.736; R2 = 0.91; Pitting (%) = 0.002 – 0.05 PPFD2 – 0.56 PPFD + 37.79; R2 = 0.894). ‘Dashehari’ mango had the highest incidence of fruit pitting (21.8%), and ‘Tommy Atkins’ had the lowest incidence (9.1%). The concentrations of leaf calcium (Ca) [1.17% (w/v) vs. 2.08% (w/v)], and boron (B) (28 µg g–1 vs. 45 µg g–1) were lower in branches bearing pitted fruit than in those bearing normal fruit, respectively. There were inverse quadratic relationships between the mean incidence of pitting and mean leaf Ca levels (Pitting (%) = 11.40 Ca2 – 48.84 Ca + 61.95; R2 = 0.48), and mean leaf B levels (Pitting (%) = 0.040 B2 – 3.67 B + 90.26; R2 = 0.97), and a linear relationship between leaf B and Ca concentrations (B = 20.93 Ca + 0.156; R2 = 0.76). These results suggest that pitting is related to low light levels, and to low concentrations of Ca and B. Experiments are in progress to resolve this problem using foliar or soil applications of Ca and B, and pruning of trees.

The fruit pitting disorder—A physiological anomaly in mango ( Mangifera indica L.) due to deficiency of calcium and boron

A new disorder known as fruit pitting has been observed in some Indian mango orchards during the recent years. In this disorder, there is a development of some sunken pits on fruit peel, which distract consumers. Based on preliminary observations, it was observed that deficiency of nutrients could be the cause, and hence systematic studies were conducted in five indigenous cultivars such as ‘Alphonso’, ‘Amrapali’, ‘Dashehari’, ‘Mallika’ and ‘Neelum’, and five exotic mango cultivars such as ‘Edward’, ‘Irwin’, ‘Rosari’, ‘Sensation’ and ‘Tommy Atkins’ with the aim to observe the fruit pitting incidence and degree, and to investigate its probable causes. Our studies indicated that nearly 13% of the mango fruit was affected by fruit pitting with variable degree and magnitude. All indigenous cultivars had higher incidence of fruit pitting than exotic cultivars. ‘Dashehari’ had the maximum incidence of fruit pitting (30.3%), followed by ‘Amrapali’ (28.6%), and ‘Rosari’ the least (3.4%). Our studies indicated that the incidence of fruit pitting in mangoes was nearly 13% with a significant variability among the cultivars ( Table 1). Although the concentrations of most of the major nutrients such as N, P, K, Mg, and minor nutrients such as Cu, Mn, Fe, Zn, did not differ significantly. However, the pitted fruit had lower Ca (1.53%) and B (22 mg kg −1) concentrations than normal fruit (2.47% and 38 mg kg −1, respectively), indicating that deficiency of Ca and B probably is the cause for fruit pitting in mangoes.

Fluorescence imaging as a non-destructive method for pre-harvest detection of bitter pit in apple fruit ( Malus domestica Borkh.)

Bitter pit in apples still causes significant losses, especially in the export markets of ‘Golden Delicious’ apples from South Africa. Orchard practices to reduce the possibility of bitter pit are followed, as well as destructive methods to predict the probability thereof, but the occurrence of bitter pit is still unacceptably high. Fluorescence imaging is a fast, non-destructive technique, able to evaluate numerous fruit within a short time span. By applying fluorescence imaging on individual fruit before any symptoms of bitter pit were apparent, lower fluorescence was shown to be associated with bitter pit development in apples in selective cases. Our results showed that, using averaged cumulative distribution functions (CDFs) of pitted and non-pitted fruit classes, it was possible to show a difference between these classes with fluorescence imaging. However, the individual distinction between all pitted and non-pitted fruit of our total sample, could not been defined as clearly. In the majority of cases, on a single fruit basis, separation in groups was not satisfactory (less than 100% accurate) based on industry requirements for a prediction technique. Results of pre-harvest imaging on apples to identify fruit with bitter pit potential at harvest showed that pitted fruit were correctly classified (75–100%). However, misclassification of non-pitted fruit (50% and less) with fluorescence imaging is still too high to be of any commercial use.

Determining the probability of bitter pit in ‘Golden Delicious’ apples through the post-harvest mineral content of individual fruit

SummaryBitter pit fruit in commercial consignments of apples still poses an economic threat to exporters from South Africa. Mineral analysis of fruit has been used with variable success to predict bitter pit prior to harvest. The possibility of increasing the accuracy of existing predictive models by using analysis of individual fruit rather than pooled samples was investigated. By improving the normality of the distributions of the different minerals and decreasing the overlap between pitted and non-pitted fruit classes, we attempted to improve the reliability of predictions based on variable threshold values. Even though our model produced a correct classification of 85% for non-pitted fruit which can be useful, this was still below the required tolerance expected on the market which, at present, is less than 2% bitter pit in an overseas consignment. The classification for pitted fruit, 63%, was not satisfactory.

NON-DESTRUCTIVE DETECTION OF PITS IN DRIED PLUMS

An economical, non-destructive device was constructed to detect pits in dried plums (prunes). The device compresses the product between a roller and a force transducer, which detects the higher force generated when a pit is present. Two methods of classifying fruit were developed, one based on the maximum magnitude of the compression force and the other on analysis of the frequency spectra of the force transducer signal during compression. The accuracy of the former was 98.6% for pitted fruit and 69.4% for fruit with pits, versus 99.1% and 75.3% for the latter. The frequency spectra classification method was more accurate, but more complex and costly to implement. The requirement to restrict the compression of the fruit to be non-destructive precludes the possibility to detect small pit fragments using this method. However, the low cost of materials for the device make it a potential method to supplement other technology currently in use to reduce the pit count in the final product.

Volatile Differences of Pitted and Non-pitted `Fallglo' Tangerine and White `Marsh' Grapefruit

Postharvest pitting, which has severely affected citrus quality, can be caused by wax application and high temperature storage. Internal volatile composition of waxed and non-waxed fruit could be an indicator of fruit susceptibility to postharvest pitting. In this study, volatile composition was compared between pitted and non-pitted `Fallglo' tangerines [Bower citrus hybrid (citrus reticulata Blanco × C. reticulata Blanco × C. paradisi Macf.) × Temple (C. reticulata Blanco × C. sinensis L.)], as well as in white `Marsh' grapefruit (C. paradisi Macf.). Pitted fruit had a higher volatile concentration than non-pitted `Fallglo' tangerines or white `Marsh' grapefruit. Concentrations of camphene, ethyl hexanoate, alpha-phellandrene, 3-carene, alpha-terpinene, p-cymene, and limonene were higher in pitted white `Marsh' grapefruit than in those of non-pitted fruit. In `Fallglo' tangerines, higher concentrations of limonene and citronellal were found in pitted fruits than in non-pitted fruit. In peel samples of grapefruit, seven different volatiles (methanol, ocimene, citronellyl acetate, alpha-copaene, trans-caryophyllene, alpha-humulene and valencene) were significantly higher in pitted peel than in non-pitted grapefruit peel. Volatiles, such as limonene could be used to predict peel disorders of white `Marsh' grapefruit and `Fallglo' tangerines during storage.

Influence of the number of calcium sprays on the distribution of fruit mineral concentration in an apple orchard

SummaryThree treatments – six, nine and twelve Ca sprays – were applied to four year old ‘Braeburn’ apple trees to establish the effect of the number of sprays on the distribution of fruit mineral concentration in the orchard. The P, K, Ca and Mg concentrations of 150 fruit and N concentration of another 150 fruit were determined two weeks before the estimated optimum harvest date and each fruit was analysed separately. At the same time a second sample of fruit was picked and stored at –0.5°C for seven weeks. After another two weeks’ storage at 17°C, fruit was examined for bitter pit development. In a second single fruit analysis the P, K, Ca and Mg concentration of 37 bitter pit and 29 bitter pit-free fruit was determined, as well as the N concentration of 27 bitter pit and 27 bitter pit-free fruit. Results of the pre-harvest analysis showed that the number of sprays affected the fruit Mg and Ca concentration, but there were no differences in N, P and K concentration between treatments. Frequency distributions of N and P were non-normal, as was the Ca distribution of the six and nine spray treatment populations. These distributions were skew with extended tails in the higher concentration class regions. This non-normality has to be taken into account when using a single mean Ca value to predict the disorder and to prevent the underestimation of the bitter pit risk. The Ca distribution of the twelve spray treatment population and the distribution of K and Mg were normal. Post-storage analysis indicated that all mineral distributions were normal, with the exception of the P distribution. Save for Ca, mineral concentrations of bitter pit and bitter pit-free fruit did not differ significantly. The bitter pit and bitter pit-free Ca distributions overlapped in the 2–3 mg 100 g–1 FW range.

Characterization, Quantitation, and Distribution of Anthocyanins and Colorless Phenolics in Sweet Cherries

The anthocyanins and colorless phenolics in 11 cultivars and hybrids of sweet cherries were characterized and quantified by high-performance liquid chromatography and gas chromatography. All of the dark-colored cherry genotypes were found to contain 3-rutinoside and 3-glucoside of cyanidin as the major anthocyanins and the same glycosides of peonidin as minor anthocyanins. Another minor anthocyanin, pelargonidin 3-rutinoside, was characterized in sweet cherries for the first time. The major colorless phenolics were characterized as neochlorogenic acid and p-coumaroylquinic acid. The total anthocyanin content ranged from 82 to 297 mg/100 g of pitted cherry for the dark cherries and from 2 to 41 mg/100 g of pitted fruit for the light-colored cherries. The neochlorogenic acid ranged from 24 to 128 mg/100 g of pitted cherry and p-coumaroylquinic acid from 23 to 131 mg/100 g, respectively. The relative amounts of the two phenolic acids varied widely across the cherry cultivars examined in this study.

Effects of water deficit on vegetative growth, fruit growth and fruit quality in Cox’s Orange Pippin apple

SummaryDegrees of soil water deficit were imposed in order to water-stress 4-year-old Cox’s Orange Pippin apple trees at two phases of growth. Water deficit, imposed from 28 November 1985 until 3 January 1986 (Phase I), reduced shoot extension by 37% compared with the fully irrigated control trees. The level of soluble solids in fruit was raised by 0.8%, the proportion of cracked fruit was increased 2-3 fold, and the incidence of bitter pit was marginally reduced, but mean fruit size and titratable acidity were not altered, when compared with fruit from the control trees. When the water deficit was imposed from 4 January until harvest on 21 February 1986 (Phase II), shoot growth was reduced by only 7%. The level of soluble solids was raised by 1%, but the incidence of bitter pit and mean fruit size were not affected. Neither of the irrigation treatments significantly reduced crop density or crop yield.

EFFECTS OF WATER IMMERSIONS ON THE INCIDENCE OF SURFACE DISORDERS IN SWEET CHERRIES

Immersion of sweet cherries (Prunus avium L. ’Van’) for 0.25 min in 0 °C water increased the incidence of pitted fruit after impact damage. Increased immersion durations up to 30 min in either 0 °C or 20 °C water had no additional significant effect on the incidence of surface pits or bruises. Significant increases in the incidence of surface pits resulted when cherries were immersed in water at 20 °C for 64 min or longer.