Influence of Md-ACS1 allelotype and harvest season within an apple germplasm collection on fruit softening during cold air storage

Previous studies have demonstrated a relationship between Md-ACS1 allelotype and apple fruit softening at ambient temperatures. The present study was undertaken to further examine the influence of this allelotype (-1/1, -1/2 or -2/2) and its interaction with harvest season (early or late) on changes in internal ethylene concentrations (IEC) and fruit softening during cold air storage. This was carried out by describing natural differences found among old apple cultivars/species and modern commercial cultivars. For late maturing cultivars, Md-ACS1-1/1 was firmer at harvest than Md-ACS1-2/2 with the heterozygote intermediate. However harvest firmness showed no differences among for the early season Md-ACS1 allelotypes. The Md-ACS1-2/2 allelotype had a slower rate of postharvest IEC increase and flesh softening compared with Md-ACS1-1/1 and -1/2 allelotypes, and late maturing cultivars had a slower rate of fruit softening than early maturing cultivars, which was independent of postharvest IEC. All three late season allelotypes and early season Md-ACS1-2/2 were firmer after storage than early season Md-ACS1-1/1 and -1/2 allelotypes, reflecting differences in both harvest firmness and softening rates. While cultivar variation in final firmness could be explained partially through Md-ACS1-mediated postharvest ethylene increases and subsequent softening, much more variation was accounted for by their differences in harvest firmness. These results are discussed in relation to strategies for breeding cultivars with superior flesh textures that are maintained during storage.     

Optimal sampling strategies for evaluating fruit softening after harvest in apple breeding

Environmental variance components associated with year, tree, and harvest date were estimated for fruit softening after harvest in apple (Malus × domestica Borkh.) to determine their relative importance and design optimum sampling strategies to discriminate genotypes in apple breeding. Fruit were stored after harvest under 20± 2 ^∘C and 80± 5%RH. Softening was evaluated by adapting the change in firmness during storage to a linear regression and defining the regression coefficient as the softening rate. Environmental variances associated with genotype × year interaction, among trees, year × tree interaction, and among harvest dates were all very small, namely, 2.7, 0.1, 5.2, and 5.7%, respectively, to the total variance obtained from the analysis of variance for the softening rate. The variance associated with genotype, at 57.3%, was very large. On the basis of the number of fruit necessary for firmness measurements, two times harvest is an efficient strategy to determine a genotype mean for the softening.     

Maturity assessment at harvest and prediction of softening in a late maturing nectarine cultivar after cold storage

The absorption coefficient μ_a measured at 670 nm in fruit pulp at harvest by time-resolved reflectance spectroscopy (TRS) has been shown to be a good maturity index for early nectarine cultivars. By including individual fruit maturity as a biological shift factor (BSF) into a kinetic model for softening it is possible to select fruit with different shelf-life potential. The BSF approach combined with TRS measurement and kinetic modeling of firmness was applied to a late maturing nectarine cultivar (‘Morsiani 90’), ripened at 20 °C after harvest or after storage at 0 °C and 4 °C, the latter conditions inducing chilling injury. At harvest the absorption coefficient μ_a had low values and low variability, indicating advanced maturity, while firmness was similar to that of early cultivars. The softening model took into account these differences, showing parameters similar to those of the early cultivars with the exception of the softening rate which was 2-6 times lower, indicating a slower softening in ‘Morsiani 90’ fruit. Decay of μ_a at 20 °C was also slower. Softening continued during storage at 4 °C, but not at 0 °C. After storage at 0 °C softening was resumed similarly to non-stored fruit, but with much variability. Fruit stored at 4 °C, which showed chilling injury, had a softening rate at 20 °C significantly higher than that of 0 °C fruit. It is suggested that the same changes in cell wall metabolism which induce the appearance of chilling injury also affect firmness and increase softening rate.     

Utilizing the IAD index to determine internal quality attributes of apples at harvest and after storage

A hand-held instrument (DA Meter, Sinteleia, Bolonga, Italy) which measures chlorophyll nondestructively and gives an I_AD index (index of the absorption difference between 670 and 720 nm), was used to measure quality attributes of three cultivars of apples. A preharvest study conducted in a ‘Starking’ apple orchard found determination coefficients of r² values of 0.79 to 0.64 between the I_AD index and starch levels, firmness, TSS and TA of fruit. Three orchards of ‘Starking’, ‘Granny Smith’ and ‘Pink Lady’ apples were sampled during commercial harvest and the apples held in air storage for up to six months. The I_AD index measured either at harvest or at removal from storage was correlated with TSS, TA and firmness of the apples. The r² values were best for TSS (0.76) and poorest for firmness (0.51). Moreover, the measurement at harvest and at removal from storage gave similar prediction coefficients for quality attributes. This measurement can be used to sort at harvest for different quality classes after storage, or at removal from storage for different marketing classes.     

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.     

Metschnikowia andauensis as a new biocontrol agent of fruit postharvest diseases

Potential antagonists were isolated from the epiphytic flora associated with oranges and pome fruit. A total of 1465 microorganisms were tested in a preliminary screening against blue and green moulds on pome and citrus fruit, respectively. Among them, approximately 3% reduced incidence and severity by more than 50% and 4 microorganisms fulfilled the selection criteria of reduction in severity and incidence by 75%. The most effective was a yeast identified as Metschnikowia andauensis, strain NCYC 3728 (PBC-2), isolated from the surface of ‘Bravo de Esmolfe’ apple fruit cultivated in North Portugal. The biocontrol activity of M. andauensis PBC-2 was dependent on its applied concentration. At 5 × 10⁶ cfu/mL incidence (% of infected wounds) and severity (lesion diameter) were reduced by 62 and 70%, respectively and at 1 × 10⁷ cfu/mL, the greatest reduction was achieved, 90% of incidence and 95% of severity. The broad spectrum of action of M. andauensis PBC-2 was evaluated with effective control being achieved against Rhizopus stolonifer, Penicillium expansum and Botritys cinerea, on ‘Rocha’ pears and on different apple cultivars and against Penicillium digitatum and Penicillium italicum on mandarins and oranges. In semi-commercial trials in cold storage, the reduction of blue mould was 90%. Rapid colonization of fresh apple fruit wounds was observed during the first 24 h of cold storage, followed by a significant population increase during the first 15 days of storage and then the population remained stable until the end of storage.     

Prediction ability of firmness decay models of nectarines based on the biological shift factor measured by time-resolved reflectance spectroscopy

The maturity of nectarines at harvest can be assessed by measuring the absorption coefficient at 670 nm (μ_a) with the non-destructive technique of time-resolved reflectance spectroscopy (TRS). A kinetic model links μ_a, converted into the biological shift factor (BSF), to firmness decrease during ripening; in this way the firmness decay model includes the variations in maturity at harvest, thereby allowing prediction of shelf-life for individual fruit. In order to study how this methodology could be practically used at the time of harvest, when μ_a can be measured non-destructively on all fruit, while the destructive measurement of firmness can only be done on a small sample, various firmness decay models were developed using either data at harvest or within 1–2 d after harvest from previous experimental research with nectarines carried out over a 5-year period. These models were then tested for prediction and classification ability by comparing the predicted firmness and class of usability to the actual ones measured during ripening and their performance compared to that of models based on data during the whole shelf-life. Our results suggest that the methodology might be used as a management tool in the nectarine supply chain. Independently from the actual softening rate, the classification at harvest based on μ_a is able to segregate fruit of different quality and maturity according to their softening behaviour during shelf-life. Among the various models, those estimated using data at harvest and after 24 h of shelf-life had better performance than those based only on data at harvest. In the 2002 and 2005 seasons, this model showed a classification ability very close to that of models based on data during the whole shelf-life. However, its performance in the 2004 season was not so good, because it could not take into account the influence of cold storage periods prior to shelf-life. All the steps necessary to apply this methodology are detailed.     

Diphenylamine and pre-slicing storage effects on the responses of apple slices to elevated CO2 atmospheres

The effect of treatment with diphenylamine (DPA) and duration of postharvest storage of whole apple fruit on the responses of fresh-cut apple slices to elevated CO₂ storage atmospheres has been investigated. On the day of harvest, ‘McIntosh’, ‘Empire’ and ‘Delicious’ apples were untreated or dipped in DPA, and were held at 0.5 °C overnight or for 6 weeks before slicing. Slices were then stored at 0, 15, 30, 45 or 60% CO₂ in 1% O₂ (balance N₂), atmospheres. Color, firmness and accumulation of acetaldehyde, ethanol and ethyl acetate of the slices were measured. Generally slices were lighter (higher L^* values) when stored in elevated CO₂ atmospheres, but atmosphere and DPA effects varied by cultivar and were affected by pre-slice storage time. Slices prepared from stored fruit were softer compared with slices prepared at harvest. Slice firmness was not affected consistently by CO₂ or DPA concentration, whether they were prepared at harvest or after storage. The effects of increasing CO₂ concentration on acetaldehyde and ethanol accumulations were variable, being affected by cultivar and storage period. DPA treatment did not affect acetaldehyde accumulation of any cultivar, or ethanol accumulation of slices prepared from fruit at harvest. However, DPA-treated ‘Empire’ and ‘Delicious’ apples stored before slicing accumulated less ethanol compared with untreated fruit. Storage of apples before processing increased the accumulation of fermentation volatile compounds by cut apples under storage atmosphere conditions.     

Control of postharvest diseases of fruit with an invert emulsion formulation of Trichoderma harzianum Rifai

Control of primary postharvest diseases caused by Rhizopus stolonifer, Botrytis cinerea, and Penicillium expansum on a variety of fresh fruit was evaluated with an invert emulsion formulation of Trichoderma harzianum. Diseases evaluated were quantified by the period of protection conferred by the antagonist and the diameter of decay lesions. Treatment of the various fruit species with formulated T. harzianum conidia in an invert emulsion significantly (P ≤ 0.05) reduced the mean lesion diameters of R. stolonifer on apple, pear, peach and strawberry, B. cinerea on grape, pear, strawberry, and kiwifruit, and P. expansum on grape, pear, and kiwifruit in comparison with the control treatment. Significant differences (P ≤ 0.05) were obtained in the mean percent reduction in lesion diameter caused by the same postharvest pathogens on the same fruit species due to the treatment with the formulated T. harzianum conidia relative to control treatment. The greatest mean percent reduction (86.7%) was obtained on apple fruit for the infection with R. stolonifer. Significant differences (P ≤ 0.05) were also obtained in the mean durations of the minimum protection period due to treatment with the formulated T. harzianum against the infection with the same postharvest pathogens on the same fruit species. The longest mean duration of the minimum protection period (up to 59 days) was obtained for unwounded apple fruit against the infection with R. stolonifer. Overall, the results indicate that the treatment with the invert emulsion formulation of T. harzianum protected fruit from infection by the primary postharvest pathogens of the fruit tested for up to 2 months and reduced the diameters of decay lesion up to 86% and is a promising treatment to prolong the postharvest shelf-life of fresh fruit.     

Identification and genetic characterization of an ethylene-dependent polygalacturonase from apricot fruit

During fruit ripening a loss of firmness occurs, which is a key factor limiting postharvest life. In apricot, Prunus armeniaca L., a wide range of fruit firmness at commercial maturity has been observed in different cultivars. Endopolygalacturonase (endoPG) activity has been reported to be associated with differences in firmness in many fruit species, but never in apricot. In this paper, we reported the identification of an apricot cDNA (PaPG) coding for an endoPG-like protein with 393 amino acids. Protein sequence comparison with known polygalacturonases (PGs) revealed that multiple features as conserved domains and functional residues and a predicted signal peptide were present in PaPG. Moreover, a phylogenetic analysis of this and other plant PGs placed PaPG into a clade containing endoPGs expressed in fruit, abscission and dehiscence zones without a propeptide sequence, very close to PRF5 from peach (Prunus persica L. Batsch). PaPG gene expression increased during postharvest storage of the fruit, correlating with fruit softening and ethylene release, and it responded to exogenous ethylene treatments. We localized the PaPG gene in apricot linkage group 4 after genetic mapping based on SNP analysis, in a position apparently syntenic to the PRF5 locus from peach. Results obtained offer genetic evidence supporting the hypothesis that PaPG and PRF5 are orthologous genes, and consequently position PaPG as a gene of interest for studies on fruit softening in apricot, and contribute to the development of molecular tools for breeding apricots with longer shelf life.     

Effect of prolonged cold storage on the sensory quality of peach and nectarine

To maintain peach and nectarine quality after harvest, low temperature storage is used. Low temperatures induce physiological disorders in peach, but the effect of cold storage on the sensory quality of the fruit before it is damaged by chilling injury syndrome remains unclear. To evaluate the cold storage effect on the sensory quality two peach cultivars (’Royal Glory’ and ‘Elegant Lady’) and two nectarines (’Ruby Diamond’ and ‘Venus’) were harvested at a standardized firmness level and subjected to quality evaluations and sensory analysis at harvest and after storage at 0 °C for 35 d. For both time points, a supplementary ripening followed such that homogeneous flesh firmness and suitability for consumption was achieved.The fruit segregation through the Durofel firmness (DF), evaluated using a non-destructively method (Durofel device), allowed the formation of a uniform group of fruit in terms of flesh firmness (FF), showing scores between 45.1 and 55.9 N. The average FF in fruit ripened immediately after harvest was 22.9 N and 25.6 N in fruit ripened after cold storage for 35 d.The “acceptability” of fruit is highly correlated with “aroma”, “sweetness”, “juiciness”, “texture” and “flavor”. Only the “acid taste” parameter had no significant correlation with “acceptability” or with the other parameters evaluated.It is possible to conclude that the sensory quality and acceptability of peach and nectarine are characteristic of each cultivar and change, depending on the time elapsed after harvest. In general, it was confirmed that nectarine cultivars have a more consistent quality than peach cultivars.     

Effect of harvest maturity and cold storage on correlations between fruit properties during ripening of apricot (Prunus armeniaca)

Apricots (Prunus armeniaca) were held in air storage at 0 °C and ripened at 20 °C, or ripened at 20 °C straight after harvest, and changes in fruit quality quantified using postharvest and sensory evaluations. Maturity at harvest significantly affected flesh firmness and other quality factors. Mealiness and gel formation only developed in fruit that had been stored at low (0 °C) temperatures. Mealiness did not develop until firmness dropped below approximately 20 N, whereas gel formation began to develop when firmness was as high as 35 N. Development of mealiness and loss of juiciness were correlated; however, slight mealiness was perceived when fruit were still considered juicy. Specific cultivar-related differences were evident in the changes in firmness and development of gel formation during and after cold storage. Fruit were less liked by the sensory panel when firmness dropped below 20 N, as juiciness decreased and mealiness and gel formation increased. Cell wall studies showed changes in yields of water-soluble and CDTA (trans-1,2-cyclohexanediamine tetraacetic acid)-soluble pectin. In fruit ripened after cold storage, mealiness and gel formation was accompanied by an increase in water-soluble pectin and an increase in CDTA-soluble pectin, whereas in apricots ripened straight after harvest, water-soluble pectin increased but CDTA-soluble pectin slightly decreased. All fruit, regardless of maturity or having chilling disorders or not, fitted the same correlation between firmness and uronic acid content of water-soluble pectin, but no pattern was evident for CDTA-soluble pectin. We concluded that the increasing solubilisation of pectin was a major feature of fruit softening in apricot, whereas the differences in CDTA-soluble pectin may reflect differences in strength of cell adhesion.     

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.     

Quality changes in sapote mamey fruit during ripening and storage

Physical and chemical changes in sapote mamey (Pouteria sapota (Jacq.) H.E. Moore and Stearn) fruit during ripening and storage at various temperatures were evaluated. Ripening was associated with flesh softening, an increase in soluble solids content (SSC), and a change in flesh color from yellow or pale pink to a dark pink or red. No changes in fruit skin color or in flesh acidity were observed as ripening progressed. Ripe fruit had 30% or higher SSC, orange or red flesh (hue angle=52; chroma=45; L=60), acidity of 6–8 mM H⁺, and flesh firmness (compression force) ≤50 N. Flesh turned brown (L* value declined) in overripe fruit. Fruit held at 27, 25, or 20°C ripened in 3.5, 5 or 7 days after harvest, respectively. Fruit kept at 10°C showed minor changes in color and firmness and a slow rate of SSC increase. Fruit stored at 10 or 15°C and then ripened at 20°C had portions of the flesh with a much higher firmness and poorer development of red color compared to other parts of the fruit. This uneven ripening was probably a result of chilling injury. The number of fruit with injury was higher at 10°C than at 15°C, and increased with storage time. The rates of fruit weight loss relative to the initial fruit weight were 0.58, 0.98 and 1.83% d⁻¹ at 10, 20 and 27°C, respectively.     

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.     

Synergistic effect of Lentinula edodes and Pichia membranefaciens on inhibition of Penicillium expansum infections

The feasibility of using a lyophilized culture filtrate (LCF) from Lentinula edodes at different concentrations to improve the efficacy of Pichia membranefaciens in controlling postharvest blue mold decay in apple fruit was investigated. Application of LCF alone was effective in inhibiting the blue mold rot in apple fruit wounds in a dose-dependent manner. Moreover, the biocontrol activity of P. membranefaciens against blue mold caused by Penicillium expansum in apple fruit could be enhanced by addition of LCF. The combination of P. membranefaciens and LCF resulted in a more effective control than individual treatment of P. membranefaciens or LCF alone. The combined treatment induced higher phenolic accumulation and the upregulation of superoxide dismutase, catalase, ascorbate peroxidase, chitinase and β-1,3-glucanase activities in fruit than LCF or yeast alone, and resulted in a lower lesion diameter and disease incidence. The use of LCF may be an effective method to improve the biological activity of P. membranefaciens, and induced host defenses appear to contribute to the control mechanism of P. membranefaciens and LCF.