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Chilling injury (CI) is a major postharvest constraint in the long-term cold storage, transportation, and distribution of japanese plums (Prunus salicina). The aim of the work was to explain the development and severity of CI in japanese plums based on the oxidative stress theory following time course analysis of enzymatic and non-enzymatic antioxidants. Changes in membrane lipid peroxidation and enzymatic and non-enzymatic antioxidative systems in japanese plum cultivar Blackamber were determined at weekly intervals during 5 weeks of cold storage at 0 °C and at 2-day intervals during poststorage simulated shelf conditions (21 ± 1 °C) for 8 days after each week of cold storage. Fruit respiration and ethylene production rates showed typical climacteric patterns after removal from cold storage and these rates were relatively high after 4 and 5 weeks compared with 0 to 3 weeks of storage. The CI symptoms first appeared after 3 weeks of cold storage after fruit had been transferred to simulated shelf conditions. The incidence and severity of CI intensified with increasing storage duration. The extent of lipid peroxidation indicated by concentration of thiobarbituric acid-reactive substances and membrane damage manifested as electrolyte leakage increased with increasing duration of storage and subsequent simulated shelf conditions. Membrane lipid peroxidation exhibited positive correlation with the severity of CI. Activities of primary antioxidant enzymes and the enzymes involved in the ascorbate–glutathione cycle were determined to explain the levels of reduced and oxidized forms of cellular redox buffers, ascorbate and glutathione. In response to chilling stress, antioxidative protection systems operated efficiently during the first 3 weeks of cold storage, but extended storage resulted in loss of ability to ameliorate increasing levels of oxidative stress. In this study, the comprehensive analyses of various metabolites and antioxidative systems explain the series of events involved in development of CI in japanese plums in support of the oxidative stress theory.
To elucidate the temporal relationship between endogenous ethylene and color development, changes in ethylene concentration, fruit color, pigments, and flavonoids were monitored at different intervals during the development and ripening of ‘Pink Lady’ apple (Malus × domestica Borkh.), commencing from 60 days after full bloom (DAFB) in 2002 to 2003, and 7 DAFB in the 2003 to 2004 season. Concentrations of chlorogenic acid, phloridzin, catechin, epicatechin, and quercetin glycosides were significantly higher (P ≤ 0.05) during the early stages (7−60 DAFB) compared with the later stages (≥60 DAFB) of fruit growth. During fruit development, concentrations of phloridzin, catechin, epicatechin, and quercetin glycosides decreased to low and stable levels. However, the concentration of anthocyanin (cyanidin 3-galactoside) increased during maturation and ripening (≥168 DAFB), coinciding with a corresponding increase in percent red blush and endogenous ethylene concentration. Positive and significant correlations were found between ethylene and color development and between ethylene and total anthocyanin, but not the other flavonoid compounds. Ethylene therefore appears to be a key factor regulating anthocyanin biosynthesis and color development in ‘Pink Lady’ apple. However, the accumulation of other classes of flavonoids such as catechin, epicatechin, phloridzin, chlorogenic acid, and quercetin glycosides seems to be regulated independently of ethylene during fruit development and ripening.
Changes in endogenous free polyamines (putrescine, spermidine, spermine) were monitored from fruit set (fruit diameter 4.6 ± 0.5 mm, wt 0.09 ± 0.05 g) until 1 week before the expected harvest time in `Kensington Pride' and `Glen' to examine their role during mango (Mangifera indica L.) fruit development. Polyamines (PAs) in the pericarp tissues (exocarp and mesocarp) were estimated throughout the fruit development period, while estimations from growing ovules were started from 41 days after fruit set (DAFS). During fruit ripening, ethylene production and endogenous free PAs in skin and pulp of `Kensington Pride' mango were also monitored. PA contents of pericarp declined between fruit set and maturity from 788 to 101 nmol·g-1 fresh weight (FW) in `Kensington Pride' and from 736.6 to 89.6 nmol·g-1 FW in `Glen' during fruit development. Spermidine (SPD) and spermine (SPM) were higher than putrescine (PUT) during the initial phase of fruit growth. The highest levels of free PAs, especially SPD and SPM, at the initial stages of fruit growth suggest a potential role during the cell division phase and not in subsequent fruit development. Ovule seems to be a rich source of PAs as evident from 2.3- and 2.7-fold higher total PAs than pericarp tissues in `Kensington Pride' and `Glen', respectively. During fruit ripening of `Kensington Pride', total PAs increased in skin and pulp tissues along with the climacteric rise of ethylene, and reached maximum levels (skin 796, pulp 314 nmol·g-1 FW) on day 4 of ripening. Skin exhibited 55.8% higher mean free PAs than the pulp. PUT dominated both in skin and pulp tissues. The simultaneous increase of ethylene and free PAs during fruit ripening suggests that their biosynthesis may not be competitive, and free PAs may have evolved as a response to increased biosynthesis of ethylene.
This research was carried out to extend the postharvest storage of japanese plum (Prunus salicina Lindl. cv. Tegan Blue), which has a short shelf life limiting its export potential. The effects of 1.0 μL·L−1 1-methylcyclopropene (1-MCP) and modified atmosphere packaging (MAP), alone or in combination, on quality of mature japanese plum fruit during storage (0 ± 1 °C and 90% ± 5% relative humidity) were investigated. The activities of enzymes of ethylene biosynthesis [1-aminocyclopropane-1-carboxylic acid synthase (ACS), 1-aminocyclopropane-1-carboxylic acid oxidase (ACO), and 1-aminocyclopropane-1-carboxylic acid (ACC) content] and those of cell wall-associated enzymes [exo-polygalacturonase (exo-PG), endo-polygalacturonase (endo-PG), pectin esterase (PE), and endo-1,4-β-D-glucanase (EGase)] were also measured. 1-MCP-treated fruit stored in normal atmosphere or in MAP had lower ACC content and inhibited ethylene production with reduced ACS and ACO activities compared with fruit stored in MAP and in normal atmosphere. Similarly, 1-MCP-treated fruit, stored either in normal atmosphere or in MAP, were firmer with reduced exo-PG, endo-PG, PE, and EGase activities compared with fruit stored in MAP and in normal atmosphere. During storage as well as during ripening, fruit stored in MAP exhibited a higher rate of respiration compared with other treatments. MAP exacerbated the effect of 1-MCP in reduction of ethylene production and fruit softening. 1-MCP application in combination with MAP after 5 and 7 weeks of storage delayed the fruit ripening by 10 and 8 days in contrast with control fruit, respectively. During storage, and as well as in ripe fruit, weight loss was reduced in fruit stored in MAP either with or without 1-MCP application. Control fruit and 1-MCP-treated fruit, stored in a normal atmosphere or in MAP, had the same values for the following parameters: chromaticity value L*, C*, and hue angle, titratable acidity, and concentrations of soluble solids, ascorbic acid, and total antioxidants. In conclusion, 1-MCP application in combination with MAP can be used effectively to reduce the ethylene biosynthesis and fruit softening during cold storage and to extend the storage life up to 7 weeks followed by 8 d of ripening without any adverse effects on the quality of ripe fruit.
Growth and maturation of `Pink Lady' (Malus domestica Borkh.) apples with special emphasis on ethylene biosynthesis and color development were monitored in Western Australia during 2002-2003. Changes in fruit growth, respiration rate, ethylene production, anthocyanin accumulation and development of red blush were evaluated between 60 days after full bloom (DAFB) and commercial harvest (191DAFB). Fruit diameter, length and fresh weight showed the typical single sigmoid growth curve, with linear increases until 158 DAFB. High respiration rate and ethylene production were recorded 60 DAFB followed by rapid decrease until 144 DAFB and then a steady increase, which peaked between 172 and 179 DAFB. Red blush on the fruit surface showed steady increase from 167 DAFB and corresponded to concomitant decrease in hue angle. Total anthocyanin increased from 167 DAFB till harvest and synchronized with increasing ethylene and maturity of apples. There were significant (P ≤ 0.001), direct linear relationships between ethylene production and total anthocyanin (r = 0.63, y = 7.6032x + 2.4756), total anthocyanin and red blush (r = 0.74, y = 0.5082x -1.54). Significant (P ≤ 0.001) negative direct linear relationships between total anthocyanin and hue angle (r = -0.89, y = -0.5973x + 110.14), and ethylene and hue angle (r = -0.69, y = -5.37x + 109.60) were recorded. Increasing anthocyanin content and red blush also coincided with decreasing daily temperatures in the orchard. Reduction in fruit firmness and acidity and increase in TSS from 167DAFB were good indicators of advancing maturity of apples. Our experimental results indicate that red blush in `Pink Lady' apples develops a few weeks before harvest and is regulated by ethylene biosynthesis and temperature.
Particle bombardment seems to be a promising method for genetic transformation of mango and the information on various factors affecting transient gene expression is inconclusive. Pro-embryonic masses (PEMs) of mango (Mangifera indica L. cv. Kensington Pride) used for particle bombardment were established and maintained in vitro using various liquid and semisolid media containing 2,4-D. Various factors affecting the biolistic transformation efficiency in mango and subsequent transient gene expression were optimized using β-glucuronidase (GUS) as a reporter gene driven by CaMV 35S promoter. Our results show that bombardment pressure significantly affects transient gene expression with the best results obtained from PEMs bombarded at 1200 kPa of helium pressure under vacuum. The application of osmoticum pre and post-bombardment also significantly increased the transient gene expression in the PEMs as compared to the controls. Mannitol (0.2 m) proved to be the best osmoticum in improving transient GUS expression as compared to sorbitol. The duration of exposure of PEMs to mannitol (0.2 m) both pre and post-bombardment also played a significant role in improving transient gene expression. The transient GUS expression was significantly highest with a pre-treatment of 0.2 m mannitol for 4 hours as compared to 0, 8 and 12 hours. The post-bombardment treatment of 0.2 m mannitol for 16 hours resulted in significantly highest transient gene expression as compared to 0, 4, 8, 12, 20 and 24 hours. In conclusion, PEMs of `Kensington Pride' mango bombarded at 1200 kPa, which were exposed to mannitol (0.2 m) for 4 and 16 hours pre and post bombardment respectively, resulted in the highest transient GUS expression (25.1 GUS foci/mg PEMs).
The effects of preharvest application of AVG and ethephon alone, or in combinations, on color development, fruit quality and shelf life were tested in `Pink Lady' apples (Malus domestica Borkh.) in Western Australia during 2002.The experiment aimed at improving color without adversely affecting fruit quality at harvest and after long term cold storage. Treatments included 124.5 g·ha-1 AVG only [148 Days after full bloom (DAFB)]; 280 g·ha-1 ethephon only (148 DAFB); AVG (148 DAFB) followed by ethephon (166 DAFB); and control. Fruit were evaluated for color development, internal ethylene concentration (IEC) and quality at commercial harvest(181DAFB) and 45, 90, and135 days after cold storage (1 °C ± 0.5 °C). At harvest, ethephon with or without AVG significantly (P ≤ 0.05) improved red blush and total anthocyanin in fruit skin. AVG+ethephon treated-fruit had higher total anthocyanin and TSS compared to AVG alone and control fruit. There were no significant differences among different AVG and ethephon treatments for fruit firmness and IEC. During different storage periods, fruit treated with AVG alone and AVG+ethephon had significantly lower IEC compared to fruit treated with ethephon only and the control, however the interactions between treatments and storage periods were not significant for fruit firmness. AVG + ethephon and ethephon alone did not significantly affect fruit color during different storage periods, which showed that the subsequent ethephon spray on AVG-treated fruit had overcome the inhibitory effect of AVG. Our experimental results showed that application of AVG followed by ethephon improved color in `Pink Lady' apples without compromising fruit quality including firmness during extended cold storage.