Immersion of fruit in water to improve radio frequency treatment to control brown rot in stone fruit

Brown rot caused by Monilinia spp. is the most important postharvest disease of stone fruit. Currently, no chemical fungicides are allowed in the European Union to be applied to stone fruit after harvest, which has increased the need to develop alternative methods. Radio frequency (RF) treatment at 27.12 MHz with fruit immersed in water was studied to control brown rot in peaches and nectarines artificially inoculated with M. fructicola. Additionally, RF treatment in air was also investigated to evaluate the benefit of water immersion to reduce the effect of fruit size on treatment efficacy. RF treatment with fruit immersed in water at 20 °C applied for 9 min significantly reduced brown rot incidence in both peaches and nectarines and no significant differences in RF efficacy were observed depending on fruit size. However, when RF treatment was applied in air for 18 min, brown rot reduction was significantly higher in large fruit than in small fruit. Finally, the decrease in exposure time of radio frequency treatment with fruit immersed in water with increasing water temperature was also studied. Reduction of treatment time to 6 and 4.5 min was achieved by increasing water temperature at 35 and 40 °C, respectively, to control brown rot without adverse external and internal damage in both ‘Baby Gold 9’ peaches and ‘Autumn Free’ nectarines.     

Continuous microwave treatment to control postharvest brown rot in stone fruit

Monilinia spp. are the most important causes of brown rot in stone fruit and no chemical fungicides are allowed in the European Union to be applied to stone fruit after harvest. From preliminary studies, microwave (MW) treatments at 17.5 kW for 50 s and 10 kW for 95 s were selected as effective conditions to control brown rot. Both treatments were investigated to control Monilinia fructicola in fruit with different weights and maturity levels and in naturally infected fruit. Fruit weight only had a significant effect on microwave efficacy in ‘Placido’ peaches treated by MW at 10 kW for 95 s in which better brown rot control was observed in small than large fruit. Maturity level did not have a significant effect on efficacy of MW treatments in any of the varieties evaluated. When both MW treatments were studied in naturally infected peaches and nectarines, brown rot incidence was significantly reduced to less than 14% compared with untreated fruit where brown rot incidence was higher than 45%. The effect of both treatments on fruit quality was also evaluated. Fruit firmness was not negatively affected in the varieties tested and even a delay of fruit softening was observed. However, internal damage around the stone was observed, especially in the smallest fruit in which high temperature is achieved at the end of both MW treatments.     

Effect of host and Monilinia spp. variables on the efficacy of radio frequency treatment on peaches

Brown rot caused by Monilinia spp. is the most important postharvest disease of stone fruit. Currently, no chemical fungicides are allowed in the European Union to be applied to stone fruit after harvest. In previous work, radio frequency (RF) treatment for 4.5 min applied with fruit immersed in water at 40 °C was very promising for the control brown rot on peaches and nectarines. In the present study, the efficacy of this radio frequency treatment was studied employing different infection times, inoculum concentrations, fruit maturity levels and in naturally infected fruit. Generally, infection time and maturity level of fruit did not have a significant effect on the RF treatment efficacy and brown rot incidence was significantly reduced in fruit inoculated 0, 24 or 48 h before treatment and at all maturity levels evaluated in both peaches and nectarines. RF treatment significantly reduced brown rot incidence at all inoculum concentrations evaluated (10³, 10⁴, 10⁵ and 10⁶ conidia mL⁻¹). However, in peaches, the treatment efficacy was slightly less when the inoculum concentration was increased to 10⁵ or 10⁶ conidia mL⁻¹. In naturally infected fruit, brown rot incidence was significantly reduced from 92% among control fruit to less than 26% in peaches and complete brown rot control was achieved in nectarines. RF treatment did not have an effect on fruit firmness in the varieties tested, and even a delay of fruit softening was observed. Moreover, both external and internal fruit appearance was not affected by the treatment.     

Impact of a decontamination step with peroxyacetic acid on the shelf-life,sensory quality and nutrient content of grated carrots packed under equilibrium modified atmosphere and stored at 7 °C

Peroxyacetic acid (PAA) is a strong oxidizer and exerts antimicrobial properties. The effect of a decontamination step with 80 and 250 mg L⁻¹ PAA on shelf-life of grated carrots stored under equilibrium modified atmospheric packaging at 7 °C was determined and compared with the shelf-life of unwashed and water-washed carrots. Microbial parameters, including total aerobic plate count, numbers of lactic acid bacteria, Lactobacillae and yeasts, and sensory quality were evaluated. Next to these parameters, atmospheric gas composition, pH and nutrient content were also monitored. The suggested packaging configuration prevented CO₂ accumulation, but at the end of the study anoxic conditions were reached for unwashed carrots and carrots washed with 80 mg L⁻¹ PAA. The microbial shelf-life of water-washed carrots was 4 d based on the yeast count, whereas the flavour was not acceptable after 5 d. The total aerobic plate count and the yeast count determined the shelf-life of carrots treated with 80 mg L⁻¹ PAA on 5 d, whereas the flavour was unacceptable after 7 d. None of the microbial parameters determined the shelf-life of carrots washed with 250 mg L⁻¹ PAA. However, this treatment had already a pronounced adverse effect on the initial sensory quality. Water washing already decreased the content of all individually studied nutrients (−16 to −28%), except for lutein content and the antioxidant capacity. Additional losses after adding PAA on day 0 were found for α-tocopherol and phenols. Regardless of the applied treatment, α- and β-carotene remained stable during storage, whereas ζ-carotene, lutein and α-tocopherol were unstable. The phenol content and the antioxidant capacity of unwashed, water-washed and 80 mg L⁻¹ PAA-treated carrots increased significantly at the end of the storage period, whereas no changes were found in carrots treated with 250 mg L⁻¹ PAA.On the condition that carrots were packed under an adequate EMA, the 80 mg L⁻¹ PAA treatment showed possibilities for extending shelf-life without pronounced effects on nutrient content.     

Control of fungal decay of apples and peaches by the biofumigant fungus Muscodor albus

The potential of the volatile-producing fungus Muscodor albus for controlling postharvest diseases of fresh fruit by biological fumigation was investigated. In vitro tests showed that M. albus volatiles inhibited and killed a wide range of storage pathogens belonging to species of Botrytis, Colletotrichum, Geotrichum, Monilinia, Penicillium and Rhizopus. Fumigation of apples for 7 days with culture of M. albus grown on autoclaved grain gave complete control of blue mold (Penicillium expansum) and gray mold (Botrytis cinerea) in wound-inoculated fruits. There was no direct contact between the fruit and the M. albus culture. Shorter fumigation times ranging between 24 and 72 h, applied immediately or 24 h after inoculation, also controlled blue mold and gray mold. In wound-inoculated peaches, 24–72 h fumigation with M. albus provided complete control of brown rot (Monilinia fructicola). The volatile profile of M. albus-colonized grain was measured by gas chromatograph connected to a flame ionization detector (GC-FID) and showed that 2-methyl-1-butanol and isobutyric acid were the major volatile compounds found in the headspace. Since M. albus is a sterile mycelium and does not require direct contact with the crops to be treated, it could be an attractive biological fumigant for controlling postharvest diseases.     

Control of Monilinia rots on fruit naturally infected by hot water treatment in commercial trials

In recent years, safer methods for the control of fruit postharvest pathogens have been investigated and heat treatment could represent an effective and safe approach for managing postharvest decay such as Monilinia rots. In the present study, the effect of hot water treatment (HWT) (60 °C for 30 and 60 s) on brown rot was investigated. More specifically, the influence of HWT was determined in in vitro trials on conidial germination of Monilinia laxa, Monilinia fructicola and Monilinia fructigena and in peach and nectarine fruit, naturally infected. The effect of hot water application on fruit quality was also assessed. M. fructicola showed a greater resistance to heat than M. laxa and M. fructigena, however conidia germination of all three species was completely inhibited by a dipping in hot water for 1 min at 55 °C. The results of a large scale experiment under commercial conditions and several pilot trials showed a good antifungal activity of HWT in naturally infected fruit. After 6 days at 0 °C and 3 days at 20 °C, in both semi-commercial and commercial trials, the inhibition of decay was higher than 78% in four trials out of six. In addition, the treated fruit showed an acceptable commercial quality and no visual damage was observed as a consequence of HWT. The results demonstrated that HWT is a promising method to control Monilinia rots of peach and nectarine, and is safe and readily available for conventional and organic production under commercial conditions.     

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).     

Characterization of sensitivity of grove and packing house isolates of Penicillium digitatum to pyrimethanil

In most northeast Argentinean citrus packing houses, postharvest fungicide treatments are based on the use of thiabendazole and imazalil. However, these fungicides have been used in a manner highly conducive to the selection and proliferation of resistant biotypes of Penicillium digitatum, the main fruit decay fungus in the area. Recently, a new fungicide, pyrimethanil (PYR), was introduced to control molds. Aims of this study were to determine the baseline sensitivities for PYR against isolates of P. digitatum considering its use in the region is not yet widespread and to evaluate the control of the fungus in vivo. One hundred and nine (109) P. digitatum isolates were collected from diseased fruit within citrus groves (43 isolates) and packing houses (66 isolates). EC₅₀ was determined for each isolate by measuring colony diameters on different agar dilutions of the fungicide. The mean EC₅₀ value of the green mold isolates collected from the groves was 0.14 ± 0.03 mg L⁻¹ while the mean EC₅₀ of those collected from packing houses was 0.13 ± 0.05 mg L⁻¹. No resistant isolates were found in the field where the fungicide is not used, while one isolate originated from a packing house showed an EC₅₀ of 3.40 mg L⁻¹, 26-fold higher than the mean level. This isolate was collected from lemons stored in cool rooms of a packing house where PYR had not been used. Fruit decay by sensitive isolates was reduced approximately 80% by PYR applied at 500–600 mg L⁻¹ by immersion for 60 s at room temperature to inoculated oranges and mandarins. In contrast, the resistant isolate was not controlled by PYR applied at 1000 mg L⁻¹. Thus, the introduction of PYR applied into packing houses should be done carefully and control strategies should be implemented in order to minimize the development of resistant isolates.     

Selection and evaluation of new antagonists for their efficacy against postharvest brown rot of peaches

During the growing seasons 2007 and 2008, 210 isolates of yeasts or yeast-like fungi were obtained from the carposphere of temperate fruit collected from organic orchards in Northern Italy. Through six rounds of in vivo screening, three isolates showing the highest biocontrol efficacy against Monilinia laxa on peaches were selected. By using molecular and morphological tools, the strain AP6 was identified as Pseudozyma fusiformata, the strain AP47 as Metschnikowia sp., and the strain PL5 as Aureobasidium pullulans. This research represents the first evidence about the potential use of P. fusiformata to control postharvest diseases of fruit. By co-culturing in vitro M. laxa in the presence of the three antagonists, neither the inactivated cells nor the culture filtrate of the three isolates had any significant effect on spore germination or germ tube elongation, allowing exclusion of the production of secreted toxic metabolites. The antagonistic activity of A. pullulans PL5 and P. fusiformata AP6 was dependent on the cell concentration. Metschnikowia sp. AP47 significantly inhibited spore germination at the three concentrations tested (10⁶ cells/mL, 10⁷ cells/mL, and 10⁸ cells/mL). The efficacy of the three strains was tested on peaches stored at three different temperatures, and their effectiveness was higher at 1 °C than at 8 °C or 20 °C. In trials carried out in semi-commercial conditions with peaches inoculated by spraying 10⁵ spores/mL of M. laxa and stored for 21 d at 1 °C and 96% RH, a cell concentration effect on the control of brown rot incidence was observed. AP6 and PL5 showed no significant differences in efficacy when applied at 1 × 10⁸ cells/mL or at 1 × 10⁷ cells/mL, indicating that they could be used at a lower concentration in potential biofungicide formulations. Finally, in an experiment in semi-commercial conditions on fruit not inoculated with the pathogen with 21 d storage at 1 °C and 96% RH, the evaluation of postharvest quality parameters, including firmness, total soluble solids, ascorbic acid content, and titratable acidity, showed that none of the three screened antagonists impaired peach quality, when applied before storage. The present study identified three antagonistic microorganisms with potential exploitation as active ingredients for the development of products for postharvest control of brown rot on peaches.     

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.     

The combination of curing with either chitosan or Bacillus subtilis CPA-8 to control brown rot infections caused by Monilinia fructicola

Recently, it has been reported that brown rot in peaches and nectarines can be effectively controlled by exposing fruit to 50 °C for 2 h and 95-99% relative humidity (RH). This treatment was effective at reducing infections that had become established in the field. However, it did not provide protection for further Monilinia fructicola infections, indicating that fruit was susceptible to subsequent infections after the treatment process and before cool storage. Chitosan and Bacillus subtilis (strain CPA-8) were evaluated for their ability to prevent M. fructicola infections and for their ability to complement the heat treatment. Two chitosan concentrations (0.5% or 1%) were applied at three temperatures (20, 40 or 50 °C) for 1 min to wounded and unwounded fruit that were artificially inoculated with M. fructicola. One percent chitosan applied at 20 °C had a preventive effect against further M. fructicola infections on heat-treated fruit that had been previously inoculated: brown rot incidence was reduced to 10%, in comparison with the control (73%). However, chitosan applied to wounded fruit had a poor preventive effect. The antagonist, B. subtilis CPA-8, had a preventive effect in controlling M. fructicola infections: the incidence of brown rot was reduced to less than 15% for both varieties evaluated (‘Baby Gold 9’ and ‘Andros’ peaches), in comparison with the control fruit (higher than 98%). In contrast, when fruit were stored at 0 °C, this preventive effect was not detected. These findings indicate that heat-treated fruit can be protected from subsequent fruit infection after heat treatment by use of chitosan or B. subtillis CPA-8, thereby providing packinghouses with an effective biologically based, combined approach to the management of postharvest brown rot.     

Control of postharvest grey mould (Botrytis cinerea Per.: Fr.) on strawberries by glucosinolate-derived allyl-isothiocyanate treatments

The vapours of allyl-isothiocyanate (AITC) were evaluated in in vitro and in vivo trials against Botrytis cinerea, a severe pathogen of strawberries. In in vitro trials AITC activity was assayed on conidial germination and mycelial growth of the fungus. The mycelium appeared less sensitive to AITC than conidia (EC₅₀ values of 1.35 mg L⁻¹ and 0.62 mg L⁻¹, respectively). In addition, AITC had a fungistatic effect against the pathogen, since the values of EC₅₀, for both parameters, increased by around 30% after AITC removal. In in vivo trials, ‘Tecla’ and ‘Monterey’ strawberries (spring-bearing and day-neutral cultivars, respectively) obtained from organic production and naturally infected by B. Cinerea, were exposed for 4 h in an atmosphere enriched by pure AITC or derived from defatted seed meals of Brassica carinata (0.1 mg L⁻¹, in a 0.1 m³ treatment cabinet). After 2 days at 0 °C and another 3–4 days at 20 °C, the fruit were evaluated for grey mould infections. The AITC treatment reduced the decay caused by the pathogen by over 47.4% up to 91.5%, significantly different from the untreated fruit. No significant differences were found between synthetic and glucosinolate-derived AITC. Residue analysis performed on fruit at the end of storage (7 d after treatment) showed values lower than 1 mg kg⁻¹. Total phenolic content and antioxidant capacity estimated in treated and untreated strawberries showed no significant difference between control and AITC treated fruit. Our results show it is possible to reduce the incidence of postharvest grey mould on strawberries with a treatment of AITC (0.1 mg L⁻¹) for 4 h, opening a potential application of biofumigation in the postharvest control of B. cinerea in strawberry.     

Effects of cinnamon extract,chitosan coating,hot water treatment and their combinations on crown rot disease and quality of banana fruit

The antifungal activities of cinnamon extract (CE), piper extract (PE) and garlic extract (GE) were evaluated on banana crown rot fungi (Colletotrichum musae, Fusarium spp. and Lasiodiplodia theobromae) in vitro. The assay was conducted with extracts of CE, PE and GE with concentrations of 0, 0.1, 0.5, 1.0, 5.0, 10.0 and 0.75 g L⁻¹ of carbendazim (CBZ) on potato dextrose agar at room temperature. CE completely inhibited conidial germination and mycelial growth of all fungi at 5.0 g L⁻¹. PE totally suppressed mycelial growth of all fungi at 5.0 g L⁻¹ and conidial germination at 10.0 g L⁻¹ except for Fusarium spp. GE had no significant effects but low concentrations (0.1 and 0.5 g L⁻¹) enhanced germ tube elongation of the three fungi. The ED₅₀ values were higher for mycelial growth than for conidia except for Fusarium spp. Combined treatments were investigated on crown rot development in banana fruit (Musa AAA group ‘Kluai Hom thong’). Treatments included 5.0 g L⁻¹ CE, 1% (w/v) chitosan solution, hot water treatment (HWT, 45 °C for 20 min), CE plus chitosan, CE plus HWT and 0.75 g L⁻¹ of CBZ, applied before and after inoculation of the fruit. Crown rot development was assessed during storage at 13 °C for 7 weeks. Disease development was least (25%) on CE treated fruit after inoculation compared to CBZ but was higher when CE was applied before inoculation. Chitosan significantly delayed ripening as in terms of peel color, firmness, soluble solids and disease severity. CE showed no negative effects on quality of fruit. CE plus HWT caused unacceptable peel browning.     

Essential oils and silver nanoparticles (SNP) as novel agents to extend vase-life of gerbera (Gerbera jamesonii cv. ‘Dune’) flowers

The aim of this study was to evaluate the efficacy of silver nanoparticles (SNP) and essential oils as novel antimicrobial agents in extending the vase-life of gerbera (Gerbera jamesonii cv. ‘Dune’) flowers. The vase-life of flowers held in a solution containing 5 mg L⁻¹ SNP plus 6% sucrose was found to be significantly higher than with 8-HQC (8-hydroxyquinoline citrate) or control treatments. However, the vase-life was not different to that of flowers held in similar concentrations of silver nitrate. All gerbera flowers held in SNP solutions showed significantly higher relative fresh weight than the control. Vase-life of gerbera flowers was extended by addition of either 50 or 100 mg L⁻¹ carvacrol and either 1 or 2 mg L⁻¹ SNP from 8.3 to 16 d. In addition, the relative fresh weight and solution uptake of gerbera flowers were increased by addition of 100 mg L⁻¹ essential oils and 1 or 2 mg L⁻¹ SNP as compared to that of control flowers. Our results suggest the potential application of essential oils or SNP as novel alternatives to common chemicals used in preservative solutions for gerbera flowers.     

Preharvest application of abscisic acid promotes anthocyanins accumulation in pericarp of litchi fruit without adversely affecting postharvest quality

Pericarp colour of litchi fruit is an important quality attribute that determines its market value and consumer acceptance. Plant growth regulators (PGR) such as abscisic acid (ABA) and ethephon are known to play important roles in peel colour development during maturation and ripening of non-climacteric fruits (e.g. grape and litchi). Our aim was to investigate the effects of preharvest application of ABA, ethephon and their combination on pericarp colour and fruit quality of litchi (cv. Calcuttia) and also to assess the potential effects on postharvest performance of fruit. Exogenous application of ABA (150 or 300 mg L⁻¹) at the colour-break stage significantly increased the concentration of total anthocyanins and cyanidin-3-O-rutinoside, the major anthocyanin contributing ∼71–96% of the total anthocyanins, in litchi pericarp compared to ethephon (500 μL L⁻¹). Among different anthocyanins quantified, the relative contribution of cyanidin-3,5-diglucoside to the total anthocyanins was significantly higher in all PGR-treated fruit compared to the control, but the concentration of cyanidin-3-O-glucoside was specifically enhanced by ABA. No significant effect on the concentrations of epicatechin, and quercetin-3-O-rutinoside was observed in response to PGR treatments. Ethephon (500 μL L⁻¹) treatment did not significantly increase the anthocyanin levels in pericarp, but it caused more degradation of chlorophyll pigments than control. Aril quality with regard to firmness, soluble solids and acidity was not significantly affected by PGR treatments, except that ethephon-treated fruit showed significant softening and lower acidity. Postharvest changes in fruit quality attributes including pericarp browning during cold storage at 5 °C for 14 d were mainly related to the storage duration effect, rather than PGR treatment. In conclusion, ABA treatment (150 or 300 mg L⁻¹) at the colour-break stage enhanced anthocyanins accumulation in litchi pericarp without adversely affecting postharvest quality and storage stability for 14 d.     

Development of SCAR markers and a semi-selective medium for the quantification of strains Ach 1-1 and 1113-5, two Aureobasidium pullulans potential biocontrol agents

Aureobasidium pullulans strains Ach 1-1 and 1113-5 are two effective biocontrol agents against Botrytis cinerea and Penicillium expansum on stored apples. In the present work, a monitoring system allowing their identification and quantification was developed. The methodology used consisted of the development of both molecular markers and a semi-selective medium. The random amplified polymorphic DNA (RAPD) technique was applied to a collection of 15 strains of A. pullulans, including Ach 1-1 and 1113-5. Five specific RAPD fragments were amplified for strain Ach 1-1 and three others for strain 1113-5. Among them, a fragment of 528 bp specific to strain Ach 1-1 (generated with the OPR-13 RAPD primer) and another one of 431 bp specific to strain 1113-5 (amplified with the OPQ-03 RAPD primer) were selected, cloned, sequenced, and used to design sequence-characterized amplified region (SCAR) primers. Three different SCAR markers were amplified: two specific to strain Ach 1-1 (189 bp and 387 bp) and one specific to strain 1113-5 (431 bp). These SCAR primers can clearly identify strains Ach 1-1 and 1113-5 among 14 strains of A. pullulans and among eight yeast strains commonly present on apple fruit surfaces. Their selectivity was also tested using DNA extracted from epiphytic microflora of the apple surface. As a semi-selective medium, PDA medium supplemented with 0.5 mg L⁻¹ euparen, 1 mg L⁻¹ sumico, 2.5 mg L⁻¹ hygromycin B, 30 mg L⁻¹ streptomycin sulphate, and 1 mg L⁻¹ cycloheximide was selected. It inhibited the development of the air microflora and appeared highly toxic for the epiphytic microflora of apple surface without altering the growth of the targeted strains Ach 1-1 and 1113-5. The combination of the semi-selective medium and SCAR markers provides a valuable monitoring tool to specifically identify and quantify A. pullulans strain Ach 1-1 and strain 1113-5 and could be used in future studies to evaluate their population dynamics under various laboratory and practical conditions.     

Thiabendazole residue loading in dip,drench and wax coating applications to control green mould and chilling injury on citrus fruit

Green mould (caused by Penicillium digitatum) is a major cause of postharvest losses in citrus. Residue loading of thiabendazole (TBZ) with application methods typically used in South African packhouses and green mould control was studied. TBZ was applied curatively and protectively in dip, drench and wax coating treatments and fruit were inoculated with a TBZ-sensitive or a TBZ-resistant isolate of P. digitatum. The dip treatments consisted of TBZ concentrations of 0–2000 μg mL⁻¹; fruit were dipped for 60 s at 22 °C at a pH of 7. Residues differed between fruit batches and ranged from 0.5 to 1.7 μg g⁻¹ at 1000 μg mL⁻¹ TBZ. Curative dip treatments almost completely controlled green mould (>96% at 1000 μg mL⁻¹ TBZ). The residue level needed for 75% curative control ranged from 0.06 to 0.22 μg g⁻¹, depending on citrus type. Protective treatments were unreliable and control varied from 17% to 97.9% at 1000 μg mL⁻¹ TBZ between fruit batches. Drench treatments consisted of exposure times of 30, 60 and 90 s with 1000 or 2000 μg mL⁻¹ TBZ. Average TBZ residues were 2.14 μg g⁻¹ for Clementine mandarin fruit and 3.50 μg g⁻¹ for navel orange fruit. Green mould control on navel orange fruit resulted in 66–92%, 34–90% and 9–38% control for curative treatments after 6 and 24 h and protective treatments, respectively, depending on fruit batch. Wax with 4000 μg mL⁻¹ TBZ was applied at 0.6, 1.2 and 1.8 L wax ton⁻¹ fruit. Chilling injury was evaluated after fruit storage at −0.5 °C for 40 days. Average TBZ residues loaded was 1.3, 1.3 and 2.7 μg g⁻¹ at the recommended 1.2 L ton⁻¹ for Satsuma mandarin, Clementine mandarin and Valencia orange fruit, respectively. Protective treatments showed lower infection levels (14–20%) than curative treatments (27–40%) for Valencia orange fruit. The same trend was observed with Satsuma (92–95% curative; 87–90% protective) and Clementine mandarin fruit (82–90% curative; 59–88% protective), but control was relatively poor. TBZ application in wax exceeded 5 μg g⁻¹ at higher wax loads (1.2 and 1.8 L ton⁻¹). Wax treatments showed a significant reduction in chilling injury; TBZ had an additive effect. TBZ resistant isolates could not be controlled.     

Assessment of blue light treatments on citrus postharvest diseases

Exposure of mature ‘Fallglo’ tangerine fruit to blue light with a photon fluence rate 40 μmol m⁻² s⁻¹ reduced symptom development of blue mold (Penicillium italicum), green mold (Penicillium digitatum), and stem end rot (Phomopsis citri) postharvest decays. Direct exposure to blue light was required to reduce decay caused by Penicillium. Blue light (40 μmol m⁻² s⁻¹) reduced in vitro fungal growth of P. italicum and P. citri. The growth of P. digitatum was more tolerant to blue light, however, the activity of fungal polygalacturonase was reduced by blue light at the intensity of 40 μmol m⁻² s⁻¹. Gas chromatography–mass spectrometry analysis identified 29 chemical constituents in flavedo oil; blue light induced only octanal accumulation. Application of octanal suppressed growth of P. italicum, P. digitatum, and P. citri in vitro. Treatment of fruit with octanal at 5 mM or 50 mM suppressed symptom development caused by Penicillium and P. citri, but discolored the peel. Inhibition of postharvest decays by blue light may be due to a combination of inhibition of fungal growth and induction of defensive responses in the host.     

Hay yield and nitrogen harvest in smooth bromegrass mixtures with alfalfa and red clover in relation to nitrogen application

An experiment was conducted in order to investigate hay yield and nitrogen harvest in binary smooth bromegrass (Bromus inermis Leyss cv. Tohum Islah) mixtures with alfalfa (Medicago sativa L. cv. Kayseri) and red clover (Trifolium pratense L. cv. Tohum Islah) in Erzurum, Turkey for 5 years between 1991 and 1995. The Hay yield, nitrogen harvest, protein concentration and land equivalent ratio (LER) in the mixtures with alternating rows of 1:1, 2:1 and 1:2 of smooth bromegrass with alfalfa and red clover were compared to those in pure legume stands without any N-fertilizer application or pure smooth bromegrass stands that received 0, 50, 100 and 150 kg ha⁻¹ N. The mixtures had no N fertilization apart from 40 kg N ha⁻¹ in the establishment year. The dry matter production in all the mixtures receiving no N fertilizer application was higher than in pure legume stands. Pure grass stands were sustained only with the application of 150 kg ha⁻¹ N. The highest hay yields were obtained from the mixtures of smooth bromegrass (Sb) with red clover (Rc) (2Rc 1Sb) (14.65 t ha⁻¹) and with alfalfa (A) (1A 1 Sb) (14.49 t ha⁻¹). Although N application increased Sb yields in pure stands, the highest yields obtained with N fertilization were still lower than the yields in the mixtures without N application. The superiority of the mixtures was also reflected by their large N harvests (e.g. 355.9 kg N ha⁻¹ in 2Rc 1Sb plots) compared to pure Rc (317.8 kg N ha⁻¹), pure A (294.3 kg N ha⁻¹) and pure Sb stands that received 150 kg N ha⁻¹. The nitrogen harvest increased in pure Sb plots as the N doses applied increased. Furthermore, the protein concentration of the hay from the mixtures (158.2–165.7 mg g⁻¹) was equal to that of the pure A stands (165.7 mg g⁻¹) and higher than that of pure Sb stands (122.9 mg g⁻¹ at 150 kg N ha⁻¹ application) although the hay from pure Rc plots had the highest protein concentration (179.3 mg g⁻¹). The LER values were also higher in the mixtures (e.g. 1.28 in 1A 1Sb and 1.28 in 2Rc 1Sb plots) compared with the pure stands. The mixture plots also had a more balanced temporal distribution of hay. The grass component was more productive in early spring, whereas the legume fractions grew better in the summer. In conclusion, for a sustainable production of high-quality hay and greater N harvests without using N fertilizers, binary mixtures of Sb with A in alternating rows (1A 1Sb) were recommended for long-purpose stands and in alternation with double red clover rows (2Rc 1Sb) for short purpose stands under similar conditions. N application could be eliminated in the grass–legume mixtures without any yield depression.     

Effects of gamma and UV-C irradiation on the postharvest control of papaya anthracnose

Anthracnose is the main postharvest disease in papaya fruit. Today, there is considerable interest on alternative methods of control to promote resistance against pathogens and supplement or replace the use of fungicides. The goal of this work was to evaluate the effects of gamma and UV-C irradiation on Colletotrichum gloeosporioides, the causal agent of anthracnose. Mycelial growth, sporulation, and conidial germination were evaluated in vitro after fungal exposition to different irradiation doses. In the in vivo assays, ‘Golden’ papaya fruit were inoculated through subcuticular injections of a conidial suspension or mycelium discs. Next, fruit were submitted to different irradiation doses (0, 0.12, 0.25, 0.5, 0.75, and 1 kGy), using Co⁶⁰ as source, or UV-C (0, 0.2, 0.4, 0.84, 1.3, and 2.4 kJ m⁻²). To check the possibility of resistance induction by irradiation, papayas were also inoculated 24, 48, or 72 h after the treatments. The fruit were stored at 25 °C/80% RH for 7 days and evaluated for incidence and rot severity. The results showed that the 0.75 and 1 kGy doses inhibited conidial germination and mycelial growth in vitro. All doses increased fungal sporulation. The 0.75 and 1 kGy doses reduced anthracnose incidence and severity, but did not reduce them when the fruit were inoculated after irradiation. All UV-C doses inhibited conidial germination and those higher than 0.84 kJ m⁻² inhibited mycelial growth. The 0.4, 0.84, and 1.3 kJ m⁻² UV-C doses reduced fungal sporulation in vitro. There was no effect of UV-C doses and time intervals between treatment and inoculation on anthracnose control and fungal sporulation in fruit lesions. Moreover, all UV-C doses caused scald on the fruit. Thus, gamma irradiation can contribute for the reduction of postharvest losses caused by anthracnose and reduce the use or doses of fungicides on disease control.