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.     

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.     

Ripening and sensory analysis of Guatemalan-West Indian hybrid avocado following ethylene pretreatment and/or exposure to gaseous or aqueous 1-methylcyclopropene

Previous reports showed that both gaseous and aqueous 1-methylcyclopropene (1-MCP) delay ripening of avocado (Persea americana Mill.), but there are no reports of the influence of 1-MCP on its sensory attributes. The objective of this study was to evaluate the effects of ethylene pretreatment and/or exposure to gaseous or aqueous 1-MCP on fruit ripening and sensory attributes of ‘Booth 7’ avocado, a Guatemalan-West Indian hybrid. Separate experiments were conducted during two seasons (2008 and 2009) with fruit harvested at preclimacteric stage in October (early season) and in November (late season). Fruit from Season 1 were exposed to ethylene (4.07 μmol L⁻¹) for 12 h at 20 °C, and stored for more 12 h at 20 °C in an ethylene-free (ethylene, <0.1 μL L⁻¹) room prior to treatment with either aqueous (1.39 or 2.77 μmol L⁻¹ a.i.) or gaseous (3.15 or 6.31 nmol L⁻¹ a.i.) 1-MCP. Ripening was monitored and firmness, respiration, ethylene production and weight loss were measured. Texture profile analysis and sensory analysis were performed on ripe fruit only (firmness, 10–15 N). Fruit from Season 2 were not exposed to ethylene pretreatment but treated only with aqueous 1-MCP 24 h after harvest. Fruit were assessed exclusively for sensory analysis when ripe (firmness, 10–15 N). Treatment with either 1-MCP formulation effectively delayed ripening from 4 to 10 d for early-season fruit, and from 4 to 6 d for late-season fruit. Higher concentrations of 1-MCP of either formulation had the greatest effect on selected pulp textural parameters of early-season fruit; the gaseous formulation had greater effect on late-season fruit quality than the aqueous formulation. In general, sensory panelists ratings of overall liking were not affected by 1-MCP treatment. Both aqueous and gaseous 1-MCP formulations delayed ripening of the Guatemalan-West Indian ‘Booth 7’ avocado without significant loss in appearance or in sensory attributes and, therefore, could be considered for use as a postharvest treatment for this hybrid.     

Treatment with 1-MCP and the role of ethylene in aroma development of mountain papaya fruit

Mountain or highland papaya (Vasconcellea pubescens) is a climacteric fruit which develops a strong and characteristic aroma during ripening. The dynamics of aroma volatile production during ripening of whole papaya fruit were analysed by headspace-SPME. The main compounds produced by the fruit were esters (aliphatic and branched) and alcohols: the most abundant esters were ethyl acetate, ethyl butanoate, methyl butanoate and butyl acetate, comprising 88% of the volatiles in fully ripe fruit; butanol was the most abundant alcohol. Among the volatiles produced, ethyl butanoate, ethyl acetate, ethyl hexanoate and ethyl 2-methylbutanoate were found to be the most potent odour compounds. During ripening of mountain papaya fruit there was an increase in the total content of both esters and alcohols. In order to clarify the role of ethylene in aroma formation, mature fruit were treated with 0.3 μL L⁻¹ of 1-MCP (16 h at 20 °C) or with 2 g L⁻¹ Ethrel, and then allowed to ripen at 20 °C. The treatment of the fruit with 1-MCP inhibited the rise in ethylene production in the fruit, while Ethrel advanced the development of the climacteric phase. Most esters identified in mountain papaya were dependent on ethylene, showing an increase in production during ripening and in response to Ethrel treatment, and a strong reduction in response to 1-MCP treatment. The data presented provide evidence that most esters produced by mountain papaya are derived from fatty acids and amino acid metabolic pathways, both of them being affected by ethylene.     

Degreening behavior in ‘Fallglo’ and ‘Lee × Orlando’ is correlated with differential expression of ethylene signaling and biosynthesis genes

Two citrus types (‘Fallglo’ and ‘Lee × Orlando’) exhibiting differential fruit degreening response when treated with ethylene were selected. Fruit were harvested at commercial maturity but at different developmental periods (Harvest I, II and III). Rate of color change was greater in ‘Fallglo’ than in ‘Lee × Orlando’ when fruit were treated with 5 μL L⁻¹ of ethylene for 24 h. After 24 h of transfer of fruit to ethylene-free storage, rate of change decreased in ‘Fallgo’ and exhibited varied response in ‘Lee × Orlando’ depending on harvest date. ‘Fallglo’ fruit from Harvests I and II were completely degreened at the end of storage for 7 d; however ‘Lee × Orlando’ were not and were green in color. No difference in seedling triple response was observed between ‘Fallglo’ and ‘Lee × Orlando’ and sequences of the four ethylene receptors were identical between them. Expression of genes involved in ethylene biosynthesis and signaling pathways were studied in flavedo to test if differences in these pathways were correlated with differential ethylene sensitivity of the citrus types. Basal levels of ACS2 and ACO expressions declined as maturity progressed, and ethylene-induced expression of ACS1 and ACO were influenced by fruit maturity. At Harvests I and II, ethylene-induced increase in ACS1 and ACO expressions and ACC levels were greater in ‘Fallglo’ than in ‘Lee × Orlando’. Ethylene treatment influenced MACC content only during Harvest I in ‘Lee × Orlando’. MACC levels were generally higher in ‘Lee × Orlando’ than in ‘Fallglo’. Expressions of ETR1 and ETR2 were ethylene responsive in ‘Fallglo’ and only ETR1 expression was ethylene responsive in ‘Lee × Orlando’. Ethylene had more impact on ETR1 expression in ‘Fallglo’ than in ‘Lee × Orlando’. Ethylene had a negative effect on ETR3 expression which was more pronounced in ‘Lee × Orlando’ than in ‘Fallglo’. Expressions of ERS1, CTR1, EIN2, EIL1 and EIL2 were not affected by ethylene in both citrus types. Expression of chlorophyllase gene and rate of total chlorophyll degradation were higher in ‘Fallglo’ than in ‘Lee × Orlando’ during ethylene treatment. Differential degreening behavior of ‘Fallglo’ and ‘Lee × Orlando’ correlated with peel maturity, and factor(s) downstream of ethylene signaling but upstream of ethylene biosynthesis play a role in the differential sensitivity.     

Internal ethylene concentrations in apple fruit at harvest affect persistence of inhibition of ethylene production after 1-methylcyclopropene treatment

Factors that affect the efficacy of 1-methycyclopropene (1-MCP) treatment of apples [Malus sylvestris (L.) Mill var. domestica (Borkh.) Mansf.] include cultivar and maturity. In this study, ‘McIntosh’, ‘Cortland’ and ‘Empire’ apples were categorized by internal ethylene concentrations (IECs) at harvest, treated with 1 μL L⁻¹ 1-MCP, and the IECs of individual fruit followed at 30 d intervals during air storage at 0.5 °C for 90 d. IECs at harvest ranged from <0.5 μL L⁻¹ to ≥100 μL L⁻¹, 51 < 100 μL L⁻¹, and 10 < 50 μL L⁻¹ for ‘McIntosh’, ‘Cortland’ and ‘Empire’, respectively. 1-MCP treatment resulted in a decrease of IECs in fruit of all cultivars by day 30 after harvest. During subsequent storage IECs remained low in fruit with <1 μL L⁻¹ at harvest, but in ‘McIntosh’, ‘Cortland’ increased in proportion to IECs at harvest, but not in ‘Empire’. The importance of initial IECs in fruit on the persistence of 1-MCP inhibition of ethylene production was confirmed in a further experiment, in which IECs in untreated and 1-MCP treated ‘McIntosh’ and ‘Empire’ apples were measured for up to 194 d. 1-MCP also decreased 1-aminocyclopropene-1-carboxylic acid (ACC) concentrations in fruit. The results of our study are consistent with the hypothesis that IEC modulates the sensitivity of climacteric fruit to 1-MCP.     

Rhodosporidium paludigenum induced resistance in Ponkan mandarin against Penicillium digitatum requires ethylene-dependent signaling pathway

This work determined if the ethylene dependent signal pathway was required for antagonist-mediated fruit defense mechanisms through investigation of disease resistance against Penicillium digitatum in Ponkan mandarin induced by 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception, and Rhodosporidium paludigenum. Blocking ethylene perception with 1-MCP resulted in an increase in ACS1, ACS2 and ACO expression, and consequently an increase in ethylene production during mechanical wounding and resistance induction. The expression of the ethylene receptors ETR1, ETR2 and ETR4 as well as ethylene response factor (ERF) were observed with similar responses to yeast and 1-MCP stimuli, with ETR3 mRNA accumulation being the most sensitive to yeast application while ERS1 was the least sensitive. When applied at concentrations greater than 500 nL L⁻¹, 1-MCP pre-fumigation significantly reduced the fruit's natural protection and R. paludigenum induced disease resistance to Penicillium decay, indicating that ethylene perception was involved in inducting disease resistance. Moreover, expression of the defensive genes CHI, β-1,3-glucanase, PAL and CIN up-regulated by yeast was inhibited to different degrees by the 1-MCP pre-treatment. This study provides evidence that the biocontrol yeast R. paludigenum increased disease resistance in Ponkan mandarin against P. digitatum infection due to ethylene and signaling pathway dependent mechanisms.     

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.     

Combined treatment of aminoethoxyvinylglycine (AVG) and 1-methylcyclopropene (1-MCP) reduces melting-flesh peach fruit softening

The effects of postharvest application of aminoethoxyvinylglycine (AVG) and 1-methylcyclopropene (1-MCP) on ethylene production and fruit quality, and thus on transportation and shelf-life, were evaluated in melting-flesh peaches. AVG (150 mg L⁻¹) significantly reduced ethylene production, and the effect was enhanced in combination with 1-MCP (1 μL L⁻¹). However, fruit treated with AVG alone softened to untreated control levels 2 d after harvest (DAH). Treatment with 1-MCP significantly reduced the rate of softening until 2 DAH, but the fruit rapidly softened thereafter, and reached untreated control levels by 4 DAH. A combination of AVG and 1-MCP significantly reduced fruit tissue softening throughout ripening. The effect of each chemical on flesh firmness indicated that 1-MCP affected fruit response in the early stages of ripening up to 4 DAH, and AVG significantly reduced softening in the latter stages from 4 to 9 DAH. Peaches treated with AVG and 1-MCP retained their ground color during ripening, but the effect of each chemical on color is unclear. The present study indicates that combined treatment with AVG and 1-MCP significantly delays the ripening of melting-flesh peaches.     

Influence of time from harvest to 1-MCP treatment on apple fruit quality and expression of genes for ethylene biosynthesis enzymes and ethylene receptors

A strong potent inhibitor of ethylene action, 1-methylcyclopropene (1-MCP) maintains apple fruit quality during storage. To understand the influence of time after harvest until 1-MCP treatment, we studied expression patterns of genes for ethylene biosynthesis enzymes and ethylene receptors in two apple cultivars, ‘Orin’ and ‘Fuji’, which differ in ethylene production. Ethylene production and expression of MdACS1, MdERS1, and MdERS2 were suppressed in all 1-MCP-treated ‘Fuji’ fruit, but in ‘Orin’, the later 1-MCP was applied after harvest, the less was the suppression of ethylene production and expression of these genes. In fruit in which 1-MCP had low efficacy (e.g., ‘Orin’ treated at 7 DAH), ethylene production and the level of MdERS1 were briefly reduced by 1-MCP treatment at 2 days after treatment, then began to increase. Since ethylene receptors negatively regulate the ethylene signalling pathway, the increased levels of ethylene production and ethylene receptors after 1-MCP treatment might reduce 1-MCP efficacy.     

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.     

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.     

Volatile profiles of ripening West Indian and Guatemalan-West Indian avocado cultivars as affected by aqueous 1-methylcyclopropene

Separate experiments were conducted with three major commercial avocado (Persea americana Mill.) cultivars grown in Florida: ‘Simmonds’ (early-season, West Indian race); ‘Booth 7’ (mid-season, Guatemalan-West Indian hybrid); and ‘Monroe’ (late-season, Guatemalan-West Indian hybrid). Fruit were harvested at preclimacteric stage and left untreated (Control) or treated 24 h after harvest with aqueous 1-methylcyclopropene (1-MCP) at 1.39 (treatment M1) or 2.77 μmol L⁻¹ a.i. (treatment M2) (75 or 150 μg L⁻¹) for 1 min at 20 °C. Whole fruit ripening was monitored at 20 °C/92% ± 3% R.H. and based on whole fruit firmness, respiration and ethylene evolution. Fruit volatiles were assessed at preclimacteric (24 h after harvest), mid-ripe (half of initial fruit firmness) and ripe maturity stages, from 100 g of chopped pulp using a purge and trap system. Untreated, firmer fruit ‘Monroe’ (268 N at harvest) ripened within 12 d of harvest while softer fruit ‘Simmonds’ (118 N) ripened within only 6 d. 1-MCP treatment extended ripening time from 33% (M1) to 83% (M2). All fruit softened normally, indicating the potential benefits of aqueous 1-MCP as a postharvest treatment for avocado when applied at these concentrations. Volatile profiles differed among the three cultivars with several compounds detected in only one cultivar, results that may contribute to a potential identification of the origin of the cultivar based on fruit volatile composition. The West Indian cultivar ‘Simmonds’ had much higher emission of hexanal (preclimacteric fruit) and cis-3-hexenal and cis-3-hexen-1-ol (ripe fruit) than the Guatemalan-West Indian hybrids ‘Booth 7’ and ‘Monroe’. On the other hand, these latter hybrids had much higher levels of alkanes than ‘Simmonds’. Treatment with 1-MCP increased emissions of alkanes during ripening of ‘Booth 7’ and ‘Monroe’. Total volatiles of avocado decreased during ripening mainly due to the significant reduction of sesquiterpenes, the main group of volatiles in all cultivars at harvest (‘Simmonds’, 53%; ‘Booth 7’, 78%; ‘Monroe’, 66%). β-Caryophyllene was the major compound at harvest, but decreased to less than 2% in ripe fruit, at which point most sesquiterpenes were not detected. Among the 10 sesquiterpenes commonly found in the avocado cultivars in this study, only α-Copaene had significantly higher emissions in mid-ripe fruit treated with the higher concentration of 1-MCP (2.77 μmol L⁻¹ a.i.), suggesting that ethylene participates in the regulation of this sesquiterpene.     

A novel technique using 1-MCP microbubbles for delaying postharvest ripening of banana fruit

This study aimed to investigate the application of microbubble technology for delaying banana ripening. A preparation of 1-MCP designed for use as a form of aqueous micro bubble (MBs) solutions was formulated. Banana fruit were immersed in 500 nL L⁻¹ of aqueous 1-MCP microbubbles (1-MCP-MBs) or fumigated with 500 nL L⁻¹ 1-MCP, then stored at 25 °C for 8 days. 1-MCP-MBs were more effective in delaying postharvest ripening than conventional 1-MCP fumigation. 1-MCP-MBs reduced the respiration rate and ethylene production compared to the control and 1-MCP fumigated fruit. Moreover, 1-MCP-MBs delayed yellowing and maintained firmness of banana fruit during storage. These results indicate that 1-MCP-MBs can be used as an alternative method for delaying the postharvest ripening of banana fruit, and its application for other commodities needs to be further elucidated.     

Response of climacteric-type guava (Psidium guajava L.) to postharvest treatment with 1-MCP

Guava (Psidium guajava L. cv. ‘Allahabad Safeda’) fruit harvested at the mature light-green stage were exposed to 300 and 600 nL L⁻¹ 1-methylcyclopropene (1-MCP) for 6, 12 and 24 h at 20 ± 1 °C, and held in either cold storage (10 °C) for 25 days or ambient conditions (25–29 °C) for 9 days. Most of the physiological and biochemical changes during storage and ripening were affected by 1-MCP in a dose dependent manner. Ethylene production and respiratory rates were significantly suppressed during storage as well as ripening under both the storage conditions depending upon 1-MCP concentration and exposure duration. 1-MCP treatment had a pronounced effect on fruit firmness changes during storage under both the conditions. The reduced changes in the soluble solids contents (SSC), titratable acidity (TA) and vitamin C content showed the effectiveness of 1-MCP in retarding fruit ripening. Vitamin C content in 1-MCP-treated fruit was significantly higher than in non-treated fruit, and those treated with 300 nL L⁻¹ 1-MCP for 6 h. The development of chilling injury symptoms was ameliorated to a greater extent in 1-MCP-treated fruit during cold storage and ripening. A significant reduction in the decay incidence of 1-MCP-treated fruit was observed under both the storage conditions. 1-MCP at 600 nL L⁻¹ for 12 h, in combination with cold storage (10 °C) seems a promising way to extend the storage life of guava cv. ‘Allahabad Safeda’ while 1-MCP at 300 nL L⁻¹ for 12 and 24 h or 600 nL L⁻¹ for 6 h, may be used to provide 4–5 days extended marketability of fruit under ambient conditions.