Characterization of ethylene biosynthesis and its regulation during fruit ripening in kiwifruit,Actinidia chinensis ‘Sanuki Gold’

Ethylene biosynthesis in kiwifruit, Actinidia chinensis ‘Sanuki Gold’ was characterized using propylene, an ethylene analog, and 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception. In fruit harvested between a young stage (66 days after pollination) (DAP) and an early commercial harvesting stage (143 DAP), 2 days of exposure to propylene were sufficient to initiate ethylene biosynthesis while in fruit harvested at commercial harvesting stage (154 DAP), 4 days of propylene treatment were required. This observation suggests that response of ethylene biosynthesis to propylene treatment in kiwifruit declined with fruit maturity. Propylene treatment resulted in up-regulated expression of AC-ACO1, AC-ACO2, AC-SAM1 and AC-SAM2, prior to the induction of AC-ACS1 and ethylene production, confirming that AC-ACS1 is the rate limiting step in ethylene biosynthesis in kiwifruit. Treatment of fruit with more than 5 μL L^?1 of 1-MCP after the induction of ethylene production subsequently suppressed ethylene production and expression of ethylene biosynthesis genes. Treatment of fruit with 1-MCP at harvest followed with propylene treatment delayed the induction of ethylene production and AC-ACS1 expression for 5 days. These observations suggest that in ripening kiwifruit, ethylene biosynthesis is regulated by positive feedback mechanism and that 1-MCP treatment at harvest effectively delays ethylene production by 5 days.     

Ethylene synthesis,ripening capacity,and superficial scald inhibition in 1-MCP treated ‘d’Anjou’ pears are affected by storage temperature

A continuing challenge for commercializing 1-methylcyclopropene (1-MCP) to extend the storage life and control superficial scald of ‘d’Anjou’ pear (Pyrus communis L.) is how to initiate ripening in 1-MCP treated fruit. ‘D’Anjou’ pears harvested at commercial and late maturity were treated with 1-MCP at 0.15 μL L⁻¹ and stored either at the commercial storage temperature −1.1 °C (1-MCP@−1.1 °C), or at 1.1 °C (1-MCP@1.1 °C) or 2.2 °C (1-MCP@2.2 °C) for 8 months. Control fruit stored at −1.1 °C ripened and developed significant scald within 7 d at 20 °C following 3–5 months of storage. While 1-MCP@−1.1 °C fruit did not develop ripening capacity due to extremely low internal ethylene concentration (IEC) and ethylene production rate for 8 months, 1-MCP@1.1 °C fruit produced significant amounts of IEC during storage and developed ripening capacity with relatively low levels of scald within 7 d at 20 °C following 6–8 months of storage. 1-MCP@2.2 °C fruit lost quality quickly during storage. Compared to the control, the expression of ethylene synthesis (PcACS1, PcACO1) and signal (PcETR1, PcETR2) genes was stable at extremely low levels in 1-MCP@−1.1 °C fruit. In contrast, they increased expression after 4 or 5 months of storage in 1-MCP@1.1 °C fruit. Other genes (PcCTR1, PcACS2, PcACS4 and PcACS5) remained at very low expression regardless of fruit capacity to ripen. A storage temperature of 1.1 °C can facilitate initiation of ripening capacity in 1-MCP treated ‘d’Anjou’ pears with relatively low scald incidence following 6–8 months storage through recovering the expression of certain ethylene synthesis and signal genes.     

Effects of chlorine dioxide treatment on respiration rate and ethylene synthesis of postharvest tomato fruit

Tomato fruit at the mature green stage were treated with ClO₂ gas in a sealed container for 12 h, and then stored at 23 °C with 85% relative humidity (RH) for 23 d. Respiration rate, respiration-related enzymes including phosphohexose isomerase (PHI), succinate dehydrogenase (SDH), and glucose-6-phosphate dehydrogenase (G-6-PDH) and 6-phosphogluconate dehydrogenase (6-PGDH), ethylene production, and the expression of LeAOX1a, LeCOX1, LeACS2, LeACS4 and LeACO1 genes were measured. The results showed that application of ClO₂ gas was effective in reducing total respiration, cytochrome pathway respiration and the expression of LeCOX1, but no significant reduction in the activities of respiration-related enzymes was observed during storage. Fruit treated with ClO₂ resulted in lower ethylene production. Furthermore, the expression of ethylene biosynthesis related genes, including LeACS2, LeACS4 and LeACO1 was reduced by the ClO₂ treatment. These results indicate that ClO₂ treatment might delay the ripening of tomato fruit, possibly by a mechanism involving suppression of respiration rate and ethylene biosynthesis.     

The addition of rosehip oil improves the beneficial effect of Aloe vera gel on delaying ripening and maintaining postharvest quality of several stonefruit

In this work Aloe vera gel (AV) alone or with the addition of 10 or 2% rosehip oil was used as fruit edible coatings in a wide range of Prunus species and cultivars: peaches (‘Roma’ and ‘B-424-16’ flat type), plums (‘Red Beauty’ and ‘Songria’), nectarine (‘Garofa’) and sweet cherry (‘Brooks’). Following treatments, fruit were stored at 20 °C for 6 days and analysed for the effect of treatments on fruit ripening and quality parameters compared with uncoated fruit (control). The addition of the rosehip oil to AV gel reduced respiration rate in all fruit, and ethylene production in the climacteric ones (peaches, plums and nectarine). In addition, all the parameters related with fruit ripening and quality, such as weight loss, softening, colour change and ripening index, were also delayed in treated compared with control fruit, the effect being generally higher when rosehip oil was added to AV, and especially in those fruit that exhibited the highest ethylene production rates (‘Roma’ and flat type peaches). Although the highest effect was obtained with AV + rosehip oil at 10%, the sensory panel detected an excess of gloss and oiliness on the fruit surface, which was considered as a negative attribute. Thus, 2% rosehip oil added to AV could be used as an innovative postharvest tool to increase the beneficial effect of AV as an edible coating, especially in climacteric fruit showing high ethylene production rates.     

1-MCP regulates ethylene biosynthesis and fruit softening during ripening of ‘Tegan Blue’ plum

To investigate the effects of postharvest application of 1-MCP on ethylene production and fruit softening, activities of ethylene biosynthesis and fruit softening enzymes were measured during postharvest ripening of plum (Prunus salicina Lindl. cv. Tegan Blue) fruit after being exposed to 1-MCP (0, 0.5, 1.0 or 2.0 μL L⁻¹) at 20 ± 1 °C for 24 h. Following the treatments, fruit were allowed to ripen at ambient temperature (20 ± 1 °C), and ethylene production in fruit, activities of ACS and ACO, ACC content and fruit softening enzymes (PE, EGase, exo-PG and endo-PG) in fruit skin and pulp were recorded at different intervals. Postharvest application of 1-MCP significantly delayed and suppressed the climacteric ethylene production with reduction in the activities of ethylene biosynthesis enzymes (ACS, ACO) and ACC content, and fruit softening enzymes (PE, EGase, exo-PG and endo-PG) in the skin as well as in pulp tissues. The reduction was more pronounced with increased concentrations of 1-MCP. 1-MCP treated fruit showed different rates of fruit softening and activities of ethylene biosynthesis enzymes in the skin and pulp tissues which warrant further investigation on regulation of gene expression related to these enzymes with the inhibitory effect of 1-MCP.     

Physiological response of ‘Larry Ann’ plums to cold storage and 1-MCP treatment

The aim of this work was to study the specific effects of low temperature and 1-MCP treatment on ethylene metabolism and oxidative behaviour in plums (Prunus × salicina cv. Larry Ann). Control fruit were stored at 20 °C or 0 °C and the 1-MCP (625 nL L^?1) treated fruit at 0 °C. Changes in the kinetics of ethylene production upon removal were related to changes in ACC metabolism (ACC and MACC levels), oxidative behaviour (H₂O₂ content) and enzymatic antioxidant potential (SOD, CAT and POX enzymes) during cold storage. Low temperature stress inhibited the synthesis of MACC, which appeared to be the basic process that regulated ACC and ethylene production at ambient temperature. Although 1-MCP treatment inhibited ethylene production and ACC accumulation in the cold, it did not inhibit the accumulation of MACC. Neither cold nor 1-MCP treatment induced oxidative stress. Nevertheless, the 1-MCP treatment significantly impaired the increase in POX activity observed during cold storage. Collectively these results showed the underlying role that ACC metabolism plays in the ripening behaviour of cold-stored plums, confirming previous results. The results also indicate that MACC and malonyl transferase activity are the key regulatory factors that control ripening and possibly some ethylene-related disorders such as chilling injury in cold-stored plums.     

Controlled atmosphere storage of guava (Psidium guajava L.) fruit

The effects of controlled atmospheres (CA) on respiration, ethylene production, firmness, weight loss, quality, chilling injury, and decay incidence of three commercially important cultivars of guava fruit were studied during storage in atmospheres containing 2.5, 5, 8, and 10 kPa O₂ with 2.5, 5, and 10 kPa CO₂ (balance N₂) at 8 °C, a temperature normally inducing chilling injury. Mature light green fruit of cultivars, ‘Lucknow-49’, ‘Allahabad Safeda’ and ‘Apple Colour’, were stored for 30 days either in CA or normal air, and transferred to ambient conditions (25–28 °C and 60–70% R.H.) for ripening. CA storage delayed and suppressed respiratory and ethylene peaks during ripening. A greater suppression of respiration and ethylene production was observed in fruit stored in low O₂ (≤5 kPa) atmospheres compared to those stored in CA containing 8 or 10 kPa O₂ levels. High CO₂ (>5 kPa) was not beneficial, causing a reduction in ascorbic acid levels. CA storage was effective in reducing weight loss, and maintaining firmness of fruit. The changes in soluble solids content (SSC), titratable acidity (TA), ascorbic acid, and total phenols were retarded by CA, the extent of which was dependent upon cultivar and atmosphere composition. Higher amounts of fermentative metabolites, ethanol and acetaldehyde, accumulated in fruit held in atmospheres containing 2.5 kPa O₂. Chilling injury and decay incidence were reduced during ripening of fruit stored in optimal atmospheres compared to air-stored fruit. In conclusion, guava cultivars, ‘Lucknow-49’, ‘Allahabad Safeda’, and ‘Apple Colour’ may be stored for 30 days at low temperature (8 °C) supplemented with 5 kPa O₂ + 2.5 kPa CO₂, 5 kPa O₂ + 5 kPa CO₂, and 8 kPa O₂ + 5 kPa CO₂, respectively.     

Respiration rate,ethylene production and biochemical variations of ackee fruit arils (Blighia sapida Köenig) stored under three temperature regimes

The storage of fruit is characterized by many physiological and biochemical changes, and this study aimed to study respiration rate, ethylene production, and other biochemical variations of ackee fruit arils (Blighia sapida), cheese variety, stored at 5, 10 and 20 °C during eight days. During storage, respiration rate decreased but ethylene production increased. Glucose, fructose, sucrose, and short chain fructooligosaccharides – 1-kestose, nystose and DP-5 – and total phenolic compounds also decreased, however, the decrease was much higher at 20 °C. The L*, a*, b, C* and H* values showed that lower temperatures preserved much better colour and visual quality, and arils stored at 5 °C were rated excellent compared to those stored at 10 and 20 °C. The quality of arils stored at 10 °C also was more than satisfactory, while arils stored at 20 °C were completely spoiled after 8 days and showed high weight losses compared to arils stored at 5 and 10 °C, which did not show any spoilage and very low weight losses. In conclusion, the results demonstrated that ackee fruit arils can be stored in very good conditions for a minimum of eight days under low temperature regimes, although at 5 °C arils showed the best shelf-life.     

Effects of postharvest treatments on gene expression in Prunus persica fruit: Normal and altered ripening

Peach (Prunus persica) fruit have a short shelf-life, and the most common method employed to delay ripening and increase their postharvest life is cold storage. However, after extended storage at low temperature some cultivars have alterated ripening processes, resulting in a lack of juice and a woolly texture. To improve our understanding of the molecular mechanisms involved in the responses of peach fruit to cold storage we determined gene expression changes of fruit (cv. O’Henry) under different postharvest conditions: ripening (5 days at 21 °C), cold storage (21 days at 4 °C) and induction of woolliness (21 days at 4 °C followed by 5 days at 21 °C).Cluster analyses of genes differentially expressed between treatments revealed unique patterns associated with biological processes that operate during postharvest treatments. Genes up-regulated during postharvest ripening and woolliness include components of ethylene, and aroma biosynthesis as well as oxidative stress response. During cold storage treatment and woolliness, several genes linked to the oxidative stress response increased in abundance, suggesting changes in redox status. Quantitative RT-PCR analysis showed a sequential increase levels of mRNAs encoding key components of cellular stress response. Moreover, after 21 days of cold storage, expression of genes encoding oxidoreductase, catalase, superoxide dismutase and gluthatione reductase was still significantly higher than before cold treatment, suggesting that fruit cells were able to respond to the increased production of ROS that was induced by extended cold storage. In the woolly fruit, up-regulation of stress response genes was accompanied by down-regulation of key components of metabolic pathways that are active during peach ripening. The altered expression pattern of these genes might account for the abnormal ripening of woolly fruit.     

Ripening behavior and quality of modified atmosphere packed ‘Doyenne du Comice’ pears during cold storage and simulated transit

The effect of MAP on extending storage life and maintaining fruit quality was studied in ‘Doyenne du Comice’ (Pyrus communis L.) pears at Hood River and Medford, Oregon. Control fruit packed in standard perforated polyethylene liners started to show senescent core breakdown and lost the capacity to ripen at 20 °C after 4–5 months of cold storage in Hood River and after 5.25–6 months in Medford. LifeSpan^® L257 MAP achieved steady-state atmospheres of 15.8% O₂ + 3.7% CO₂ in Hood River and 15.7–17.5% O₂ + 3.8–5.7% CO₂ in Medford. MAP inhibited ethylene production, ascorbic acid degradation and malondialdehyde accumulation, and extended storage life for up to 6 months with maintenance of fruit flesh firmness (FF) and skin color without commercially unacceptable level of physiological disorders. After 4, 5 and 6 months at −1 °C, MAP fruit exhibited climacteric-like patterns of ethylene production and softened to proper texture with desirable eating quality on day 5 during ripening at 20 °C. After 6 months at −1 °C plus 2 weeks of simulated transit conditions, MAP fruit maintained FF and skin color and had good eating quality at transit temperatures of 2 and 4.5 °C (10.1–11.5% O₂ + 4.8–5.2% CO₂), but reduced FF substantially and developed internal browning disorder at 7.5 and 20 °C (3.2–7.2% O₂ + 7.9–9.5% CO₂). The storage life of ‘Doyenne du Comice’ pears with high eating quality could be increased by up to 2 months when packed in MAP as compared with fruit packed in standard perforated polyethylene liners.     

Role of putrescine in regulating fruit softening and antioxidative enzyme systems in ‘Samar Bahisht Chaunsa’ mango

The role of putrescine (PUT) in regulating fruit softening, antioxidative enzymes and biochemical changes in fruit quality was investigated during ripening and cold storage of mango (Mangifera indica cv. Samar Bahisht Chaunsa). Fruit were treated with various PUT concentrations (0.0, 0.1, 1.0 and 2.0 mM) and were allowed to ripen at 32 ± 2 °C for 7 days, or stored at 11 ± 1 °C for up to 28 days. Respiration rate and ethylene production were measured daily during ripening and cold storage. Cell wall degrading enzymes such as exo-polygalacturonase (exo-PG), endo-polygalacturonase (endo-PG), pectin esterase (PE), endo-1,4-β-d-glucanase (EGase), antioxidative enzymes including superoxide dismutase (SOD), peroxidase (POX), and catalase (CAT), fruit firmness as well as biochemical fruit quality characteristics were estimated during ripening and cold storage at 2 and 7 day intervals, respectively. PUT treatments reduced respiration rate, ethylene production and maintained higher fruit firmness during ripening as well as cold storage. PUT-treated fruit exhibited significantly suppressed activities of cell wall enzymes (exo-, endo-PG and EGase), but retained higher PE activity during ripening and cold storage. Total phenolic and antioxidant contents were significantly higher in PUT-treated fruit during ripening as well in the cold storage period than in the controls. Activities of antioxidative enzymes (CAT, POX and SOD) were also significantly higher in PUT-treated fruit during ripening as well as cold storage. SSC and SSC:TA were lower in PUT-treated fruit, while TA and ascorbic acid content showed the reverse trend. In conclusion, pre-storage 2.0 mM PUT treatment inhibited ethylene production and suppressed the activities of cell wall enzymes, while resulting in higher activities of antioxidative enzymes and maintaining better fruit quality during ripening and cold storage.     

The use of electrical impedance spectroscopy to assess the physiological condition of kiwifruit

The electrical impedance of kiwifruit [Actinidia deliciosa (A. Chev) C.F. Liang et A.R. Ferguson, cv. Hayward] was studied during fruit ripening. Measurements were made on whole fruit, and tissues excised from the outer pericarp, inner pericarp and core. Alternating current at frequencies between 50 Hz and 1 MHz was passed through fruit and tissue samples, and complex impedance spectra were separated into the resistances of the apoplast, cytoplasm and vacuole, and capacitances of the plasma membrane and tonoplast. The differences in R_50 Hz and R_1 MHz between tissues (representative of apoplast resistance and total tissue resistance, respectively) were explained in terms of the anatomy and composition of the respective tissues. Some variations were seen from one year to the other. During ripening, there was little change in the impedance characteristics of the fruit, despite a 10-fold decrease in firmness. This was unexpected since previous studies with nectarine, persimmon and tomato fruit have shown a considerable reduction in impedance during ripening. The failure to observe any impedance change was checked using a number of different methods for measuring impedance, by two different laboratories, and confirmed by measuring electrolyte leakage from tissue discs. All the results suggested that the mobility of electrolytes within the cell wall did not change during kiwifruit ripening. We speculate that physicochemical interactions that take place within the cell wall may have a major impact on the impedance of kiwifruit tissue.     

Mode of action of abscisic acid in triggering ethylene biosynthesis and softening during ripening in mango fruit

The role of abscisic acid (ABA) in triggering ethylene biosynthesis and ripening of mango fruit was investigated by applying ABA [S-(+)-cis,trans-abscisic acid] and an inhibitor of its biosynthesis [nordihydroguaiaretic acid (NDGA)]. Application of 1 mM ABA accelerated ethylene biosynthesis through promoting the activities of ethylene biosynthesis enzymes (1-aminocyclopropane-1-carboxylic acid synthase, ACS; 1-aminocyclopropane-1-carboxylic acid oxidase, ACO) and accumulation of 1-aminocyclopropane-1-carboxylic acid (ACC), enhanced fruit softening and activity of endo-polygalacturonase and reduced pectin esterase activity in the pulp. The activities of ethylene biosynthesis and softening enzymes were significantly delayed and/or suppressed in the pulp of NDGA-treated fruit. The ABA-treated fruit had higher total sugars and sucrose as well as degradation of total organic acids, and citric and fumaric acids compared with NDGA treatment. These results suggest that ABA is involved in regulating mango fruit ripening and its effects are, at least in part, mediated by changes in ethylene production.     

Effect of 1-MCP treatment and N2O MAP on physiological and quality changes of fresh-cut pineapple

The effects of both 1-MCP treatment of pineapples and packaging of their fresh-cut products with an alternative modified atmosphere (MA: 86.13 kPa N₂O, 10.13 kPa O₂ and 5.07 kPa CO₂) on physiological and quality changes of these minimally processed products were investigated. Fresh-cut fruit treated or not with 1-MCP were packed in Air or in MA and were stored at 4 °C for 10 d. The following parameters were monitored during storage: ripening index; O₂, CO₂ and C₂H₄ in the package headspace; firmness and colour. Microbial spoilage of MP pineapple samples was also investigated and a mathematical model based on the Zwietering modified Gompertz equation was used to obtain growth parameters of mesophilic bacteria, yeasts and moulds.The results showed that 1-MCP treatment and MAP in a N₂O enriched atmosphere had a positive combined effect on the inhibition of respiration and ethylene production of fresh-cut pineapple and on its softening delay, confirming previous findings about 1-MCP and N₂O preservative effects on fresh-cut fruit quality. This combined effect was not extended to the ripening index and colour maintenance, as MAP at 86.13 kPa of N₂O did not add any benefit to that of the 1-MCP treatment. From a microbiological point of view, N₂O MAP extended the shelf-life of the products of 3–4 d by increasing the lag phase of microbial growth.     

Residual effects of low oxygen storage of mature green fruit on ripening processes and ester biosynthesis during ripening in bananas

Mature green banana (Musa sapientum L. cv. Cavendish) fruit were stored in 0.5%, 2%, or 21% O₂ for 7 days at 20 °C before ripening was initiated by ethylene. Residual effects of low O₂ storage in mature green fruit on ripening and ester biosynthesis in fruit were investigated during ripening for up to 6 d at 20 °C. Concentrations of ethanol in mature green fruit did not change during storage in both 21% and 2% O₂ atmospheres, but increased in fruit stored in 0.5% O₂. The activities of alcohol dehydrogenase (ADH) in 2% and 21% O₂ atmospheres remained very low throughout the storage period, but significantly increased with 0.5% O₂. After transferring fruit to regular air and trigging ripening with ethylene, yellowing of peel, fruit softening and hydrolysis of starch in fruit stored in low O₂ atmospheres were slower than in the control. Fruit stored in low O₂ also showed a delayed onset of the climacteric peak. The activities of ADH were lower in the low O₂ stored fruit than in the control fruit. Productions of ethyl acetate, isoamyl acetate, and isobutyl acetate were remarkably suppressed by low O₂ storage. Alcohol acetyltransferase activity increased gradually with storage time in all treatments, being significantly lower in fruit with low O₂ pretreatments. The results indicate that low O₂ plus room temperature storage can extend storage life of bananas with the sacrifice of a low production of ester volatiles.