Control of blue mold of apple by combining controlled atmosphere,an antagonist mixture,and sodium bicarbonate

‘Golden Delicious’ apples were wound-inoculated with Penicillium expansum and treated with various combinations of sodium bicarbonate and two antagonists (Metschnikowia pulcherrima, Cryptococcus laurentii), and then stored in air or controlled atmosphere (CA = 1.4 kPa O₂ and 3 kPa CO₂) for 2 or 4 months at 1 °C. The antagonists survived and their populations increased during both air and CA storage. The antagonists alone reduced blue mold but were more effective when combined. Sodium bicarbonate tended to reduce lesion size when used with these antagonist, either when they were used alone or combined. Storage under CA conditions also increased the effectiveness of both antagonist, when used alone or in combination. The only treatment that completely eliminated P. expansum-incited decay was the combination of the two antagonists and sodium bicarbonate on fruit stored under CA conditions. The proper combination of alternative control measures can provide commercially acceptable long-term control of fruit decay and could help reduce our dependency on fungicides.     

Mechanisms of action and efficacy of four isolates of the yeast Metschnikowia pulcherrima active against postharvest pathogens on apples

The mechanisms of action and efficacy of four isolates (GS37, GS88, GA102, and BIO126) of the yeast Metschnikowia pulcherrima against Botrytis cinerea, Penicillium expansum, Alternaria sp., and Monilia sp., all postharvest pathogens of apple fruit, were studied in vitro and on apples, in controlled and semi-commercial conditions. An application of a cell suspension (10⁸ cells per ml) of the antagonists in artificial wounds of apples reduced growth of B. cinerea and P. expansum after storage at 23 °C. A complete suppression of the pathogen was obtained against Monilia sp., stored at 23 °C, and against B. cinerea and P. expansum, stored at 4 °C. The results against Alternaria sp. were more variable. Applications of culture filtrates and autoclaved cells of the isolates were ineffective in reducing the diameter of the lesions on the fruit, supporting the hypothesis that living cells are necessary for biocontrol. In experiments of antagonism in vitro, on different solid substrates, a reduction in the mycelium growth of the pathogens resulted, so that, at least in vitro, the antagonists could produce some diffusible toxic metabolites. In co-cultivation in vitro on a synthetic medium, B. cinerea spore (10⁵ ml⁻¹) germination was completely inhibited by the presence of 10⁸ cells of the antagonists, while culture filtrates and autoclaved suspensions were not able to reduce germination. Dipping boxes of apples cv. Golden delicious in a suspension of 10⁷ antagonist cells per ml and storing for 8 months in controlled atmosphere at 1 °C, showed levels of control against B. cinerea and P. expansum similar to those from thiabendazole.     

Effects of preharvest nitric oxide treatment on ethylene biosynthesis and soluble sugars metabolism in ‘Golden Delicious’ apples

In order to examine the influence of preharvest nitric oxide (NO) treatment on ethylene biosynthesis and soluble sugar metabolism in ‘Golden Delicious’ apples, apple trees were sprayed with 50 μM sodium nitroprusside (SNP) (a donor of NO) 14 days before harvest. The results indicated that preharvest SNP treatment can increase the NO content and the NOS activity in apple fruit, therefore, delay the accumulation of ethylene due to its inhibition on the activities of 1-aminocyclopropane-1-carboxylate synthase (ACS) and 1-aminocyclopropane-1-carboxylate oxydase (ACO). Fructose is the main sugar in ‘Golden Delicious’ apple. The synthesis of sucrose was stimulated and the decomposition of sucrose was inhibited by this treatment, thus causing the accumulation of sucrose. We can draw a conclusion that pre-harvest SNP (50 μM) treatment can increase the NO content of fruit during storage, while higher NO content can further regulate fruit ripening through its effect on ethylene and sugar metabolism in ‘Golden Delicious’ apple fruit during storage at 18 °C.     

Relationship of IAD index to internal quality attributes of apples treated with 1-methylcyclopropene and stored in air or controlled atmospheres

The research was conducted to evaluate the relationship between I_AD index (index of absorption difference between 670 and 720 nm) values and internal quality attributes of apples treated with 1-methylcyclopropene (1-MCP) and stored in air and controlled atmospheres (CA). Apples of ‘8S6923’ (Aurora Golden Gala™), ‘Fuji’ and ‘Royal Gala’ were tested. The results with Aurora Golden Gala™ show that I_AD index values were maintained at higher levels if the fruit were stored in CA and that 1-MCP had no significant effect on retaining at-harvest values. The I_AD values correlated with chlorophyll a content in the peel (R² = 0.95, P < 0.0001), but not with chlorophyll b content, internal ethylene levels, firmness or titratable acidity. ‘Royal Gala’ apples showed a similar response to Aurora Golden Gala™ apples, showing no correlation between I_AD index values and internal quality attributes of those apples when treated with 1-MCP and/or CA. In contrast, ‘Fuji’ apples showed a relationship between I_AD index value changes and internal ethylene concentrations (R² = 0.67, P < 0.05) and titratable acidity changes (R² = 0.89, P < 0.01), but not firmness. These results suggest that when 1-MCP and/or CA are applied to apples after harvest, that I_AD index values do not consistently correlate to any internal quality attributes other than peel chlorophyll a content.     

Inhibition of browning on the surface of apple slices by short term exposure to nitric oxide (NO) gas

Freshly cut slices of apple (Malus x domestica Borkh cv. Granny Smith) were fumigated with nitric oxide (NO) gas at concentrations between 1 and 500 μl l⁻¹ in air at 20 °C for up to 6 h followed by storage at 0, 5, 10 and 20 °C in air. Exposure to nitric oxide delayed the onset of browning on the apple surface with the most effective treatment being fumigation with 10 μl l⁻¹ NO for 1 h. While nitric oxide inhibited browning in slices held at all temperatures, it was relatively more effective as the storage temperature was reduced with the extension in postharvest life over the respective untreated slices increasing from about 40% at 20 °C to about 70% at 0 °C. In a smaller study on ‘Royal Gala’, ‘Golden Delicious’, ‘Sundowner’, ‘Fuji’ and ‘Red Delicious’ slices stored at 10 °C, 10 μl l⁻¹ NO for 1 h was found to be effective in inhibiting surface browning in all cultivars.     

Sensitivity of Penicillium expansum to diphenylamine and thiabendazole and postharvest control of blue mold with fludioxonil in ‘McIntosh’ apples

Penicillium expansum is one of the most important pathogens that cause blue mold in stored apples. Due to the development of resistance to the postharvest fungicide, thiabendazole (TBZ), an increase in blue mold has been observed in apple storages. The sensitivity of three TBZ-sensitive and three TBZ-resistant isolates of P. expansum to diphenylamine (DPA), scald inhibitor, was tested in vitro. Of the 94 isolates, collected later in the storage season, 41% were found resistant to both DPA and TBZ. To manage the fungicide resistance, a reduced-risk fungicide, fludioxonil, was tested against blue mold caused by TBZ-sensitive and -resistant P. expansum on ‘McIntosh’ apples treated with or without 1000 μg ml⁻¹ of diphenylamine. Fruit were assessed for disease and scald incidence during storage. Diphenylamine controlled scald in treated fruit. Higher disease incidence of blue mold was observed in apples treated with diphenylamine and low concentrations of fludioxonil (3, 5, and 75 μg ml⁻¹). DPA neither positively nor negatively affected the control of blue mold when DPA was applied together with 150, 300 and 600 μg ml⁻¹ of fludioxonil during 12 weeks of storage at 4 °C.     

NIR spectroscopy for the optimization of postharvest apple management

Apples can be stored for long time under controlled temperature and atmosphere conditions, and therefore, non-destructive and rapid tools are required to assess fruit quality and to monitor changes during the postharvest period. The aim of this study was to evaluate the feasibility of NIR spectroscopy to optimize postharvest apple management and to follow changes in fruit quality during storage. An FT-NIR system operating in diffuse reflectance in the range 12,500–3600 cm⁻¹ was used to evaluate the physico-chemical (dry matter, soluble solids, colour and firmness) and some nutraceutical characteristics (total phenolics, total flavonoids and antioxidant activity) of ‘Golden Delicious’ apples, which were stored for about six months at 1 °C in controlled atmosphere, over two subsequent years. Spectral data were elaborated by PLS regression and LDA classification techniques. Good correlation models between spectral data and chemical and physical parameters were obtained for soluble solids, a* colour coordinate and firmness (0.81 < R² < 0.90 in calibration and 0.79 < R² < 0.89 in cross validation). Even higher correlation values (0.89 < R² < 0.95 in calibration and 0.86 < R² < 0.92 in cross validation) were obtained for indexes correlated to the antioxidant capacity of apples. The classification technique Linear Discriminant Analysis was applied to spectral data, in order to discriminate apples on the basis of storage time. Average correct classification was higher than 93% in validation and close to 100% in calibration, indicating high potential of NIR spectroscopy for the estimation of storage time of apple lots.     

Tolerance of codling moth,and apple quality associated with low pressure/low temperature treatments

A combination of low pressure (LP) and low temperature (LT) may serve as a phytosanitary disinfestation treatment for fresh fruit. In this study, different life stages of codling moth (eggs, 2nd to 3rd instar larvae, 5th instar larvae and pupae) were treated in hypobaric chambers maintained at 10 °C and 1.33 kPa with nearly saturated humidity (>98%). Weight loss, color, firmness, titratable acidity (TA), and soluble solids content (SSC) were selected as quality parameters to evaluate the quality changes of ‘Red Delicious’ apples before and after the LPLT treatment. Results showed that the 5th instar larvae were the most tolerant life stage for codling moth under LPLT treatment conditions. Insect mortality increased with increasing LPLT treatment time to >98% after 12 days of exposure to 10 °C temperature and 1.33 kPa pressure. Although stored in less than optimum conditions for apples, the measured quality variables of ‘Red Delicious’ were maintained relatively well after 15 days of LPLT treatment. The results suggest that LPLT technology has potential as an alternative, non-chemical disinfestation treatment for apples.     

Increasing maturity reduces wound response and lignification processes against Penicillium expansum (pathogen) and Penicillium digitatum (non-host pathogen) infection in apples

Penicillium expansum is the main postharvest pathogen of pome fruit and is a necrotrophic fungus that requires wounds to infect the fruit. Therefore, injuries caused during harvest and postharvest handling provide an optimal locus for infection. In this study, the effect of wound response in apples harvested at three different maturity stages and stored at two different temperatures (20 and 0 °C) infected with P. expansum (pathogen) and Penicillium digitatum (non-host pathogen) was evaluated. The effect of wounding and pathogen inoculation on lignin content was also quantified. At 20 °C, less decay incidence and severity were observed when time between wounding and inoculation increased, and these differences were more important in fruit from immature and commercial harvests. However, at 0 °C, wound response was too slow to prevent P. expansum infection. Lignin content was highest in fruit from the immature harvest. Our results indicated that maturity and storage temperature play an important role in apple wound response. This is the first report demonstrating that P. digitatum, a non-host pathogen, was able to develop rots in over-mature apples.     

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.     

Combination of peracetic acid and hot water treatment to control postharvest brown rot on peaches and nectarines

Brown rot caused by Monilinia spp. is the most important postharvest disease of stone fruit. From preliminary studies, the combination of 0.25% hydrogen peroxide, 0.02% peracetic acid (PAA) and 0.075% acetic acid, corresponding to 300 mg L⁻¹ of PAA, was selected to control Monilinia fructicola. Brown rot control was similarly controlled when the same concentration of PAA was applied with a PAA-based commercial product. In order to reduce PAA concentration, combinations of different concentrations and temperatures were evaluated. A treatment of 200 mg L⁻¹ of PAA at 40 °C for 40 s was selected to control pre-existing and future infections, different inoculum concentrations of M. fructicola and to control brown rot on naturally infected fruit. Brown rot was completely controlled with the selected treatment when peaches and nectarines were inoculated 0 h before the treatment but it was not controlled when infection time was increased to 24, 48 and 72 h. Also, the treatment significantly controlled brown rot at all inoculum concentrations evaluated (10³, 10⁴, 10⁵ and 10⁶ conidia mL⁻¹) in both peaches and nectarines, but no protection against future infections was observed. In naturally infected fruit, brown rot incidence was slightly but significantly reduced to 61 and 36% in ‘Roig d’Albesa’ and ‘Placido’ peaches, respectively, but not in nectarines. Immersion for 40 s in 200 mg L⁻¹ of PAA at 40 °C provides an alternative treatment to control only recent infections of Monilinia spp. whatever their concentration without generally affecting fruit quality.     

Effects of repeated 1-methylcyclopropene (1-MCP) treatments on ripening and superficial scald of ‘Cortland’ and ‘Delicious’ apples

Postharvest 1-MCP can maintain fruit quality and inhibit development of superficial scald, a physiological storage disorder found in apple fruit, but the extent of the inhibition can vary by cultivar. In this study, we investigated whether multiple applications of 1-MCP, which are now permitted by a label modification of the commercial 1-MCP product, SmartFresh™, might improve scald control. ‘Cortland’ and ‘Delicious’ apples were untreated, treated on the day of harvest with the antioxidant inhibitor of scald, diphenylamine (DPA), or with 1 μL L⁻¹ 1-MCP at different intervals after harvest. Treatment times (days) were 1, 4, 7, 1 + 4, 4 + 7, 1 + 4 + 7, 7 + 14, 7 + 28, 7 + 42, and 7 + 84. Internal ethylene concentrations (IECs), flesh firmness, and accumulations of α-farnesene and conjugated trienols (CTols) were measured at harvest, at the time of treatment, and at intervals during air storage at 0.5 °C for up to 36 weeks. Scald was completely inhibited by DPA and all 1-MCP treatments in ‘Delicious’. However, effective control of scald in ‘Cortland’ was obtained with 1-MCP treatments within the first 4 days of harvest, either alone or in combination. Scald control with delayed 1-MCP treatments resulted in poorer scald control that was comparable to that obtained with DPA. IECs and α-farnesene accumulation were similar in untreated and DPA treated fruit, but inhibited by 1-MCP. However, differences among 1-MCP treatments became more evident with increasing storage periods. Inhibition of IECs and α-farnesene accumulation was greater in fruit treated on days 1, 4, 1 + 4, 4 + 7, 1 + 4 + 7, than on day 7 alone. A second application of 1-MCP on day 14 to fruit treated on day 7 increased inhibition of IECs, α-farnesene and CTol accumulations, but increasing delays before the second 1-MCP treatment resulted in progressively less inhibition of these factors. Similar effects of treatment on IECs, α-farnesene and CTol accumulations were found for both cultivars, even though no scald was detected in treated ‘Delicious’ apples. The results indicate that initial 1-MCP treatments should be applied to faster ripening cultivars such as ‘Cortland’ within a few days of harvest.     

Responses of 1-MCP application in plums stored under air and controlled atmospheres

The potential of 1-MCP for controlling ripening in ‘Angeleno’ plum fruit under air and controlled atmosphere (CA) storage was explored, and the possibility that 1-MCP can inhibit development of brown rot caused by Monilinia laxa and internal breakdown in ‘Fortune’ and ‘Angeleno’ plums tested. After harvest, fruit were exposed to 300 and 500 nl l⁻¹ (in 2003) and 500 nl l⁻¹ 1-MCP (in 2004) at low temperatures (0–3 °C) for 24 h. After treatment the plums were stored in air at 0 °C and ‘Angeleno’ fruit were also stored in CA storage (1.8% O₂ + 2.5% CO₂). Following storage, fruit were kept at 20 °C. In ‘Angeleno’ fruit, 1-MCP was effective in delaying the loss of firmness and colour changes during holding at 20 °C. 1-MCP reduced brown rot in fruit stored in CA but no significant reduction was found in air storage. Internal breakdown, a major physiological storage disorder in plums, was inhibited by 1-MCP treatment. Furthermore, since 1-MCP applied in air storage showed better results than the control in CA conditions, an application of 1-MCP before air storage could be the best way to reduce the ripening process for short or medium storage periods (40 and 60 days). CA storage plus 1-MCP treatment could be used for long periods (80 days).     

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.     

Wound response in orange as a resistance mechanism against Penicillium digitatum (pathogen) and P. expansum (non-host pathogen)

Penicillium digitatum is the most devastating postharvest pathogen of citrus. In addition, Penicillium expansum is the main pathogen of pome fruit, although recent studies have demonstrated its ability to infect oranges under some conditions. In this study, we evaluated wound response in ‘Valencia’ oranges harvested at three different maturity stages and the effect of wound response on the establishment of both pathogens when fruit were stored at two different temperatures (20 and 4 °C). The effect of wounding and pathogen inoculation on lignin content, was also quantified. Lastly, the expression of several phenylpropanoid pathway-related genes was also analyzed by semi-quantitative RT-PCR. Results indicated that, in general, P. digitatum exhibited lower decay incidence and severity as time between wounding and inoculation increased. Decay incidence and severity were higher in fruit from the over-mature harvest than in fruit from immature and commercial harvests. P. expansum was able to infect fruit at 20 °C but lesions were small compared to lesion size of fruit stored at 4 °C. Lignin content in wounded fruit (control) and in samples wounded and inoculated with P. expansum was highest in fruit from the immature harvest at 7 d post-wounding and inoculation. Wounded fruit had higher expression of pal1, comt1 and pox1 genes at 48 h than at 24 h. However, samples inoculated with P. digitatum showed lower expression at 48 h than at 24 h. Our results indicated that maturity and storage temperature play an important role in orange wound response.     

Physicochemical measurements in ‘Mondial Gala®’ apples stored at different atmospheres: Influence on consumer acceptability

Standard quality parameters, consumer acceptability, emission of volatile compounds and ethylene production of ‘Mondial Gala^®’ apples (Malus × domestica Borkh.) were determined in relation to storage atmosphere, storage period and shelf-life period. Fruit were harvested at the commercial date and stored in AIR (21 kPa O₂:0.03 kPa CO₂) or under three different controlled atmospheres (CAs): LO (2 kPa O₂:2 kPa CO₂), ULO1 (1 kPa O₂:1 kPa CO₂), or ULO2 (1 kPa O₂:2 kPa CO₂). Fruit samples were analysed after 12 and 26 weeks of storage plus 1 or 7 d at 20 °C.Apples stored in CA maintained better standard quality parameters than AIR-stored fruit. The volatile compounds that contributed most to the characteristic aroma of ‘Mondial Gala^®’ apples after storage were butyl, hexyl and 2-methylbutyl acetate, hexyl propanoate, ethyl butanoate, ethyl hexanoate, ethyl, butyl and hexyl 2-methylbutanoate. Data obtained from fruit analysis were subjected to principal component analysis (PCA). The apples most accepted by consumers showed the highest emission of ethyl 2-methylbutanoate, ethyl hexanoate, tert-butyl propanoate and ethyl acetate, in addition to the highest titratable acidity and firmness values.     

Control of Neofabraea alba by plant volatile compounds and hot water

Fumigation by plant volatile compounds and hot water treatment were tested in vitro and in vivo for their activity against Neofabraea alba (anamorph Phlyctema vagabunda), the cause of lenticel rot in apple fruit. In vitro trials with volatile compounds showed a consistent inhibition of pathogen growth by carvacrol, trans-cinnamaldehyde, citral and trans-2-hexenal, while (?)-carvone, hexanal, p-anisaldehyde, 2-nonanone and eugenol showed progressively lower inhibition. The greatest inhibition of mycelial growth was demonstrated by carvacrol (effective doses for 50 and 95 inhibition [ED₅₀ and ED₉₅] = 5.9 and 17.0 μL L^?1, respectively; minimum inhibitory concentration [MIC] = 36.9 μL L^?1) and of conidial germination by trans-2-hexenal (ED₅₀ and ED₉₅ = 4.1 and 6.9 μL L^?1, respectively; MIC = 9.2 μL L^?1). Hot water showed a complete inhibition of conidial germination in vitro after 10, 2 and 1 min of exposure at 40, 45 and 50 °C, respectively, and a complete inhibition of mycelial growth after 20 min of exposure at 75 °C. Among the volatile compounds tested, only 25 μL L^?1 of carvacrol slightly reduced fungal infection on artificially infected apples (11.4% efficacy). Hot water treatment at 45 °C for 10 min showed high efficacy in the control of lenticel rot on apples. Reduction of infection was 80% in artificially inoculated fruit (cv Golden Delicious) and 90% in naturally infected fruit (cv Pink Lady) after 90 and 135 d of storage, respectively.