Application of low concentrations of ozone during the cold storage of table grapes

The control by ozone of postharvest decay of table grapes, caused by Botrytis cinerea and other pathogens, was evaluated in chambers and commercial storage facilities. Ozone at 0.100 μL/L or higher inhibited the spread of gray mold among stored grapes. Ozone diffusion into many types of commercial packaging was measured. Boxes made of uncoated paper corrugate inhibited diffusion more than those composed of coated paper corrugate, plastic corrugate, hard plastic, or expanded polystyrene. Internal packaging of hard plastic clamshell containers inhibited diffusion less than low density polyethylene cluster bags. Atmospheres of 0.100 μL/L ozone in the day and 0.300 μL/L at night reduced the natural incidence of gray mold by approximately 65% after 5–8 weeks of storage. Its effectiveness to control postharvest decay was compared to sulfur dioxide fumigation. After 68 days at 1 °C the incidence of gray mold among grapes stored in air, ozone, or with weekly sulfur dioxide fumigation was 38.8%, 2.1%, and 0.1%, respectively. However, decay by other fungi, such as Alternaria spp. and Penicillium spp., was controlled by sulfur dioxide, but not by ozone. In some tests, rachis appearance was moderately harmed by ozone. The combination of ozone use in storage following a single initial sulfur dioxide fumigation, or its use in between biweekly sulfur dioxide fumigations, controlled both gray mold and other pathogens and matched the commercial practice of initial and weekly sulfur dioxide fumigation. The use of both gases in this way reduced sulfur dioxide use greatly. Differences in flavor of grapes treated with ozone were not detectable compared to those stored in air, and grapes treated with ozone were preferred over those treated with sulfur dioxide.     

Effect of continuous 0.3 μL/L gaseous ozone exposure on fungicide residues on table grape berries

The persistence of residues of some fungicides, commonly applied in table grape vineyards to reduce bunch rot, was investigated during the cold storage of ‘Thompson Seedless’ table grape stemmed berries in atmospheres of air or 0.3 μL/L ozone enriched air. Grape berries were sprayed with a mixture of boscalid, iprodione, fenhexamid, cyprodinil, and pyrimethanil solutions, dried in air for 24 h, and packed in plastic clamshell containers in expanded polystyrene boxes. The boxes were stored either in ozone or in ambient air atmosphere (2 °C, 95% RH) for 36 d. Residue analyses were done initially and at 12-d intervals using gas chromatography-mass spectrometry. Residues of boscalid, iprodione, fenhexamid, and pyrimethanil declined during storage in air, but cyprodinil residues did not change significantly during 36-d storage. Storage in the ozone atmosphere markedly accelerated the rates of decline of fenhexamid, cyprodinil, and pyrimethanil, but not those of boscalid or iprodione. At the end of storage, degradation of fenhexamid, cyprodinil, or pyrimethanil was 1.6-, 2.8-, or 3.6-fold higher, respectively, in the ozone atmosphere compared that in air. Despite their structural similarity, pyrimethanil declined more rapidly in an ozone atmosphere than cyprodinil. Fenhexamid declined in both air and ozone more rapidly than the other fungicides; at the end of storage period, only 59.2% or 35.5% of the initial residue remained after air or ozone storage, respectively. Our results have shown that gaseous ozone treatment during storage has a great potential for degrading contemporary fungicides related to table grape production.     

Mode of action of nitric oxide in inhibiting ethylene biosynthesis and fruit softening during ripening and cool storage of ‘Kensington Pride’ mango

The mode of action of nitric oxide (NO) in inhibiting ethylene biosynthesis and fruit softening during ripening and cool storage of mango fruit was investigated. Hard mature green mango (Mangifera indica L. cv. ‘Kensington Pride’) fruit were fumigated with 20 μL L⁻¹ NO for 2 h at 21 °C and allowed to ripen at 21 ± 1 °C for 10 d, or stored at 13 ± 1 °C for 21 d. During ripening and cool storage, ethylene production and respiration rate from whole fruit were determined daily. The 1-aminocyclopropane-1-carboxylic acid (ACC) content, activities of ACC synthase (ACS), ACC oxidase (ACO), and fruit softening enzymes such as pectin esterase (PE), endo-1,4-β-d-glucanase (EGase), exo- and endo-polygalacturonase (exo-PG, endo-PG) as well as firmness and rheological properties of pulp were determined at two- and seven-day intervals during ripening and cool storage, respectively. NO fumigation inhibited ethylene biosynthesis and respiration rate, and maintained higher pulp firmness, springiness, cohesiveness, chewiness, adhesiveness, and stiffness. NO-fumigated fruit during cool storage and ripening had lower ACC contents through inhibiting the activities of both ACS and ACO in the fruit pulp. NO-fumigated fruit showed decreased activities of exo-PG, endo-PG, EGase, but maintained higher PE activity in pulp tissues during ripening and cool storage. In conclusion, NO fumigation inhibited ethylene biosynthesis through inhibition of ACS and ACO activities leading to reduced ACC content in the fruit pulp which consequently, reduced the activities of fruit softening enzymes during ripening and cool storage.     

Biocontrol of Botrytis cinerea in table grapes by non-pathogenic indigenous Saccharomyces cerevisiae yeasts isolated from viticultural environments in Argentina

Botrytis cinerea, the causal agent of gray mold, is an important disease of grapes. Yeasts are members of the epiphytic microbial community on surfaces of fruits and vegetables and because some yeasts inhibit fungi they are used as biocontrol agents. The major objective of the present work was to isolate yeasts from grapes, vineyard soil, and grape must and select them for their ability to prevent gray mold onset after harvest. Yeasts that were found effective against the fungus were also assayed for their possible pathogenicity in humans. Two antagonism experiments were performed to study the effect of yeasts on B. cinerea, an in vitro study with Czapeck Yeast Extract Agar and an in vivo study with grape berries at 2 °C and 25 °C; both experiments were conducted at different yeast concentrations (10⁵, 10⁶ and 10⁷ cfu/mL). Antagonists were subsequently assayed for their ability to colonize and grow in fruit wounds. The biocontrol yeasts were also examined for their possible pathogenicity in humans: phospholipase and proteolytic activity, growth at 37 °C and 42 °C, pseudohyphal formation and invasive growth. A total of 225 yeasts belonging to 41 species were isolated from must and grape berries and 65 of them, representing 15 species, exhibited in vitro inhibition of B. cinerea at 25 °C. These 65 biocontrol yeasts were subsequently assayed in vivo and 16 of them (15 Saccharomyces cerevisiae and 1 Schizosaccharomyces pombe) showed antagonistic properties against B. cinerea at 25 °C. Only one isolate (S. cerevisiae BSc68) was able to inhibit mycelial growth of B. cinerea on grape berries at both 2 °C and 25 °C. The biomass of this strain in grape wounds increased 221.5-fold at 25 °C after 3 d and 325.5-fold at 2 °C after 10 d of incubation. An increase in the concentration of certain yeasts significantly enhanced their antagonistic activity. All yeast isolates determined as biocontrol agents under in vivo conditions were isolated from fermenting musts. Twelve biocontrol agents (S. cerevisiae) revealed one or more phenotypical characteristics associated with pathogenicity in humans but none of them showed all characteristics together. The fact that there exist few reports on S. cerevisiae and none on Sch. pombe as biocontrol agents against B. cinerea makes our results even more relevant.     

Structural and physiological changes associated with the skin spot disorder in apple

Skin spot is an important physiological disorder of ‘Elstar’ apples (Malus × domestica Borkh.) that occurs after fruit have been removed from controlled atmosphere storage. Skin spots are irregular patches of small, round, brown blemishes. Cross-sections reveal a browning of protoplasts (coagulated) and of cell walls that extends into the hypodermis. Skin spots are always associated with linear, gaping and non-gaping microcracks in the cuticle. Staining of apple skin with calcofluor white usually results in white fluorescence of cell walls but, within a skin spot, the white fluorescence is weak or absent. Cell walls within, and in the immediate vicinity of skin spots stain with phloroglucin/HCl indicating the presence of lignin. The area of skin affected by skin spots was positively and linearly correlated with the area of the non-blush fruit surface infiltrated by acridine orange. In general, skin spots were limited to the non-blush fruit surface and occurred more frequently near the stem-end than the calyx region of the fruit. Skin spot areas were correlated with a 2.5-fold increase in water vapour permeability compared with unaffected areas (23.8 ± 4.0 m s⁻¹ with skin spots, 9.6 ± 2.1 × 10⁻⁵ m s⁻¹ without skin spots). Strips of the fruit skin from regions with skin spots had an increased maximum force and modulus of elasticity. Dipping fruit in ascorbic acid (0.1 or 0.3 mM for 10 min) before storage decreased the area affected by skin spots. There was no effect of dipping in ethanol/water (70%, v/v, 15 min) or in solutions of captan (1.5 g L⁻¹, 10 min) or trifloxystrobin (0.1 g L⁻¹, 10 min). In contrast, prestorage treatment with 1-methylcyclopropene (630 nL L⁻¹ for 24 h) or poststorage incubation in H₂O₂ (10% for 2, 6, 10 and 16 h) increased skin spots. Our data are consistent with a typical cell response to an oxidative burst that seems to be focussed on particular regions of the ‘Elstar’ fruit surface by concentrations of cuticular microcracks, and that is possibly caused by reoxygenation injury upon removal from CA storage.     

Pheophytinase activity and gene expression of chlorophyll-degrading enzymes relating to UV-B treatment in postharvest broccoli (Brassica oleracea L. Italica Group) florets

Pheophytinase (PPH) activity and gene expression of chlorophyll (Chl)-degrading enzymes relating to UV-B treatment in postharvest broccoli (Brassica oleracea L. Italica Group) florets were determined. PPH is involved in the dephytylation of Mg-free Chl a, pheophytin (Phy) a. However, in vitro chlorophyllase (Chlase, EC.3.1.1.14) also uses Phy a as a substrate to produce pheophorbide (Pheide) a by dephytylation. For an accurate determination of PPH activity, the PPH protein fraction was separated from Chlase protein by ammonium sulfate precipitation. The protein precipitated by 45-60% saturated ammonium sulfate included a little bit of Chlase activity and was suitable for PPH determination. PPH activity in broccoli florets treated with a UV-B dose of 19 kJ m⁻² was repressed for the first 2 d of storage at 15 °C, whereas it increased gradually with senescence of control broccoli florets. The expression level of BoCLH1 was reduced in broccoli florets on day 4 of storage, while BoCLH2 and BoCLH3 were up-regulated with UV-B treatment. A high BoPAO expression level was found in senescent broccoli florets, and the up-regulation of this gene was delayed by UV-B treatment. The highest expression level of BoPPH was found in the control, and its expression was clearly repressed by UV-B treatment on day 2 of storage. We suggest that the up-regulation of Chl-degrading enzyme genes could be delayed by UV-B treatment, resulting in the suppression of floret yellowing in stored broccoli.     

Germination of fungal conidia after exposure to low concentration ozone atmospheres

The germinability of conidia of Alternaria alternata, Aspergillus flavus, Aspergillus niger, Penicillium digitatum, Penicillium expansum, or Penicillium italicum was determined periodically during exposure for approximately 100 d to a humid atmosphere of air alone or with 150 nL/L ozone at 2 °C. Conidia were exposed on glass coverslips that were removed from chambers at intervals of one week and the germination of 100 conidia of each species was assessed after incubation for 24 h on potato dextrose agar. The period in d when 50% or 95% (ET₅₀ and ET₉₅, respectively) could not germinate and 95% confidence intervals for these estimates were made using Finney's probit analysis. ET₅₀ and ET₉₅ estimates were approximately one month and two to three months, respectively. Some natural mortality of the conidia occurred during these periods, so the entire decline in germinability was not solely due to ozone. The age of the culture from which conidia were collected influenced their susceptibility to ozone. Conidia were harvested from 7, 14, 21, and 28 d old potato dextrose agar cultures of P. digitatum and exposed to 13,000 nL/L ozone at 2 °C. After 48 h of exposure to ozone, none of the conidia from the seven-day old culture germinated, while 30–35% of conidia from 14, 21, or 28 d in age cultures germinated.     

Effects of UV-C treatment on inactivation of Escherichia coli O157:H7, microbial loads, and quality of button mushrooms

This study investigated the effects of ultraviolet-C (UV-C) light applied to both sides of mushrooms on microbial loads and product quality during storage for 21 d at 4 °C. Microflora populations, color, antioxidant activity, total phenolics, and ascorbic acid were measured at 1, 7, 14 and 21 d of storage. Additionally, the inactivation of Escherichia coli O157:H7 by UV-C was determined. Results showed that UV-C doses of 0.45-3.15 kJ m⁻² resulted in 0.67-1.13 log CFU g⁻¹ reduction of E. coli O157:H7 inoculated on mushroom cap surfaces. UV-C radiation also reduced total aerobic plate counts by 0.63-0.89 log CFU g⁻¹ on the surface of mushrooms. Although mushrooms treated with UV-C had more severe browning with increasing dosage after initial treatment, the control mushrooms also browned as indicted by lower L* and higher a* values after 21 d of storage at 4 °C. In addition, the UV-C treatments apparently inhibited lesion development on the mushroom surface. During the first 7 d, irradiated mushrooms had lower antioxidant activity, total phenolics, and ascorbic acid content compared to non-radiated samples. However, irradiated mushrooms reached similar amounts of these nutrients as the control after 14 d of storage at 4 °C. In summary, UV-C radiation could potentially be used for sanitizing fresh button mushrooms and extending shelf-life.     

Cold pre-treatment is effective for 1-MCP efficacy in ‘Tsugaru’ apple fruit

1-Methylcyclopropene (1-MCP) treatment maintains apple fruit quality during storage, but its efficacy is dependent on a number of conditions. ‘Tsugaru’ apples are a major early season cultivar in Japan, but because ‘Tsugaru’ fruit produce abundant ethylene, they have a short shelf-life, and efficacy of 1-MCP is not as high with ‘Tsugaru’ as with other cultivars. To improve 1-MCP efficacy, ‘Tsugaru’ fruit were pre-cooled at −1 °C or −3 °C for 24 h before 1-MCP treatment. Ethylene production decreased with the cold treatment, resulting in better storage after 1-MCP treatment. Although ethylene production was low at the end of 24 h of the cold pre-treatment, expression of ACS1, the ethylene receptor genes ERS1, ETR1(a), ETR1b, ETR2 and ETR5, and the cell wall degradation-related gene PG1 all increased with a 24 h cold treatment. It is assumed that these elevated gene expression levels were not caused by ethylene, but more directly by cold stimulus. Thus, a short period of cold stimulus suppresses ethylene production, but induces expression of some genes. 1-MCP treatment was more effective with some initial fruit chilling.     

Chilling injury in mango fruit peel: Cultivar differences are related to the activity of phenylalanine ammonia lyase

In mango (Mangifera indica) cv. Nam Dok Mai fruit, stored at 4 °C, peel browning occurred within 9 d, while no browning was found in cv. Choke Anan fruit stored at 4 °C for 30 d. During 6 d of shelf life at 27-28 °C, following various periods of low temperature storage, the peel browning in cv. Nam Dok Mai (if not yet maximal) became worse, whereas little browning was observed in cv. Choke Anan fruit. The pulp of the fruit of both cultivars did not show browning during the 4 °C storage, but the pulp of cv. Nam Dok Mai exhibited some browning during shelf life if the fruit had been stored at 4 °C for more than 18 d. Peel and pulp color were not correlated with total free phenolics. A high correlation coefficient was observed between peel browning and PAL activity in the peel, while a very low correlation was found with peel catechol oxidase activity. The browning in the pulp was not correlated with the measured enzyme activities. The data therefore show a relation between PAL activity in the peel and low temperature-induced peel browning.     

Root growth of maize in an Italian ryegrass living mulch studied with a non-destructive method

The use of living mulch (LM) systems for maize (Zea mays L.) cultivation may reduce soil erosion and nitrate leaching. However, the yield of maize cultivated in LM is lower than in conventional farming systems. This decrease has often been attributed to belowground competition, but the lack of a suitable method to demonstrate such competition has prevented further investigation. A recently developed method allows for the direct and non-destructive observation of root growth in LM. Maize expressing a green fluorescence protein was grown in monoculture or together with Italian ryegrass (Lolium multiflorum Lam.). Root growth was screened in minirhizotrons from the time of sowing to the anthesis of the maize.Compared with the maize cultivated without Italian ryegrass, the cultivation in the LM significantly (p < 0.05) reduced the shoot dry matter (56%), leaf area (39%) and mean root density (41%) of the maize at anthesis. An analysis of variance showed that the reduction due to the living Italian ryegrass in the root density of the maize was not significantly different from the reduction in leaf area. Similarly, the reduction in the root density and shoot dry matter of the maize were not significantly different. The roots of the LM maize grew in the soil where the LM was removed but also where the LM continued growing. This study constitutes the first direct quantification of root growth and distribution in a LM system.     

Metabolite content of harvested Micro-Tom tomato (Solanum lycopersicum L.) fruit is altered by chilling and protective heat-shock treatments as shown by GC-MS metabolic profiling

The primary aim of this study was to identify metabolites associated with chilling tolerance that was engendered by a heat-shock treatment of tomato fruit pericarp (Solanum lycopersicum L. cv. Micro-Tom). Harvested mature-green fruit were immersed in 20 or 40 °C water for 7 min (‘Heat-Shock’) and then stored at 2.5 °C for 0 or 14 d (‘Chilled’). A reduction in chilling injury symptoms (i.e., slow or abnormal ripening, increased ion leakage, and increased respiration following chilling) was used to select this heat-shock treatment as optimal. Using GC-MS (Gas Chromatography-Mass Spectrometry) metabolite profiling, 363 analytes were detected in fruit pericarp of which 65 are identified metabolites. Principal Component Analysis of these data led to distinct groups among the samples based on their treatments; ‘Chilled’ and ‘Chilled + Heat-Shocked’ fruit were markedly different from each other, while the ‘Non-Chilled Control’ and ‘Heat-Shocked’ fruit were similar and grouped closer to the ‘Chilled + Heat-Shocked’ fruit. These results indicate that the heat treatment provided protection from chilling in part by altering levels of fruit metabolites. The levels of arabinose, fructose-6-phosphate, valine and shikimic acid appear to be associated with this heat-shock induced chilling tolerance since their levels were altered in the ‘Chilled’ samples (p < 0.05), relative to the control and the heat-shocked protected fruit. We also describe the metabolites we identified that could be further studied as being indicative of incipient chilling injury in mature-green tomato fruit.     

Chilling-injury of harvested tomato (Solanum lycopersicum L.) cv. Micro-Tom fruit is reduced by temperature pre-treatments

Heat-shocks were used to reduce the development of chilling injury symptoms during ripening of tomato fruit (Solanum lycopersicum L. cv. Micro-Tom). Mature green tomatoes were immersed in 30-50 °C water for 3-9 min before being chilled at 2.5 °C for 0, 0.5, 1, 2, 3, or 14 days, and then held at 20 °C for an additional 7-14 days. The affect of both heat-shock and chilling treatments were independent of fruit weight. Measured at 20 °C after 14 days of chilling, fruit exposed to 40 °C for 7 min exhibited reduced chilling injury symptoms, as measured by their advanced ripening score and decreased rate of ion leakage into an isotonic 0.2 M mannitol solution. Reduced rates of leakage from the symplastic compartment probably contributed to the 2-fold decrease in the amount of ions in the apoplastic space, when compared to the control. A subsequent paper will report the results of metabolic profiling of Micro-Tom tomato fruit subjected to treatments that significantly decreased their development of chilling injury symptoms.     

1-MCP extends the storage and shelf life of mangosteen (Garcinia mangostana L.) fruit

Mangosteen (Garcinia mangostana L.) fruit were harvested when the peel (pericarp) was light greenish yellow with scattered pinkish spots. Fruit were exposed to 1 μL L⁻¹ 1-methylcyclopropene (1-MCP) for 6 h at 25 °C and were then stored at 25 °C (control) or 15 °C. The 1-MCP treatment only temporarily delayed softening of the fruit flesh, during storage. Storage life, defined as the time until the pericarp was dark purple, was much longer in fruit stored at 15 °C than in fruit stored at 25 °C. It was also longer in 1-MCP treated fruit (storage life at 15 °C: control 18 d, 1-MCP-treated fruit 27 d). The 1-MCP treatment also increased the length of shelf life, defined as the time until the pericarp turned blackish purple or showed calyx wilting, at 25 °C. 1-MCP treatment reduced ethylene production. It also reduced pericarp levels of 1-aminocyclopropane-1-carboxylic acid (ACC), and the pericarp activities of ACC synthase (ACS) and ACC oxidase (ACO). In the fruit flesh, in contrast, 1-MCP did not affect ACC levels and ACS activity, but the treatment reduced ACO activity. Taken together, both the storage life and the shelf life of the fruit were extended by the 1-MCP treatment. A decrease in ACO activity largely accounted for the effects of the 1-MCP on ethylene production in the pericarp.     

Interference between red kidneybean (Phaseolus vulgaris L.) cultivars and redroot pigweed (Amaranthus retroflexus L.)

Field experiments were conducted in 2006 and 2007 to evaluate the competitive ability of bush type red kidneybean (RKB) (Phaseolus vulgaris L.) cultivars against redroot pigweed (Amaranthus retroflexus L.). Three cultivars of RKB (Akhtar, Sayyad and D81083) and five A. retroflexus densities (0, 4, 8, 16 and 32 plants m⁻²) were established in a factorial arrangement. A. retroflexus had a greater plant height and growth rate (GR) but a lower leaf area index (LAI) than RKB cultivars in almost all treatments. Higher densities of A. retroflexus increased LAI and GR but decreased yield of RKB cultivars. The cv. Sayyad and D81083 had the greatest and lowest LAI and GR, respectively, in competition with A. retroflexus. The maximum intercepted photosynthetically active radiation (PAR) at noon by A. retroflexus was 90.4 and 66.0% in competition with cv. D81083 and Sayyad, respectively. The seed yield and pod number per plant of RKB cultivars decreased severely with increasing A. retroflexus density. A. retroflexus seed number m⁻² was the highest and lowest in competition with cv. D81083 and Sayyad, respectively. The competitive ability of RKB cultivars was compared using parameters estimated through two-parameter yield loss-relative leaf area model. The relative ranking of the RKB cultivars examined for their competitiveness, supported by modeling results, was Sayyad > Akhtar > D81083.     

Effect of 1-methylcyclopropene (1-MCP) on reducing postharvest decay in tomatoes (Solanum lycopersicum L.)

1-Methylcyclopropene (1-MCP as SmartFresh™ technology), an ethylene antagonist, was evaluated for affecting postharvest decay caused by Alternaria alternata, Botrytis cinerea, and Fusarium spp. on ‘Quality 23’ and ‘Seminis 35’ tomatoes at green or pink stages. Fruit with natural or artificial infection were subjected to 1-MCP at 0.0 μL L⁻¹, 0.6 μL L⁻¹ for 12 h, and 1.0 μL L⁻¹ for 6 h. After 31-42 d storage, disease incidence and severity of individual diseases in 1-MCP treated fruit was significantly reduced compared with that of the untreated controls, except in one inoculated test for ‘Quality 23’ where severity of Alternaria rot in 1.0 μL L⁻¹ treated fruit were significantly higher than that of the untreated control. Fruit treated with 1-MCP at 1.0 L⁻¹ for 6 h also had significantly higher incidence of Alternaria rot in the inoculated ‘Quality 23’ and in the non-inoculated ‘Seminis 35’ compared with the fruit treated with 1-MCP at 0.6 μL L⁻¹ for 12 h. The results of this study indicate that 1-MCP can reduce postharvest decay within a certain storage period.     

Use of 1-methylcylopropene in cyclodextrin-based nanosponges to control grey mould caused by Botrytis cinerea on Dianthus caryophyllus cut flowers

Botrytis cinerea is one of the pathogens resulting in the heaviest commercial losses in ornamental cut flowers, and the severity of grey mould disease partly depends on the presence of ethylene in the storage environment. The efficacy of a β-cyclodextrin-based nanosponge 1:8 (CD-NS) - 1-methylcyclopropene (1-MCP) complex was evaluated as a novel control agent in protecting carnation (Dianthus caryophyllus L. ‘Idra di Muraglia’) cut flowers against B. cinerea infection. Two concentrations of this non-volatile 1-MCP formulation (CD-NS complex, 0.25 and 0.5 μL L⁻¹, a.i.) were compared with commercial gaseous 1-MCP (0.25 μL L⁻¹, a.i.), and an inoculated control. A non-inoculated control was also used to assess the natural infection level. Eleven days after inoculation, the development of grey mould on carnation was significantly reduced (59.9% of flower surface) in cut stems treated with the CD-NS complex at low dosage, compared to the high dosage of the CD-NS complex (91.5%), the commercial gaseous 1-MCP formulation (76.2%) and the inoculated control (100.0%). Endogenous ethylene production was associated with symptom development. Results showed a reduced ethylene production in 1-MCP treated flowers (0.25 μL L⁻¹, a.i., both suspended and gaseous formulation). The CD-NS complex could therefore be an effective alternative to conventional chemicals to protect ornamental cut flowers.     

Effects of ethylene, pollination, and ethylene inhibitor treatments on flower senescence of gentians

Flower senescence of the potted gentian (Gentiana scabra) ‘Shinbisei’ was investigated in relation to ethylene sensitivity and production. ‘Shinbisei’ flowers were used for all experiments except for those with inflorescences. Exposure to ethylene at 0.5 μL L⁻¹ or higher concentrations for 24 h markedly accelerated flower senescence, indicating that G. scabra flowers are highly sensitive to ethylene. Treatment with 0.2 or 0.5 mM silver thiosulfate complex (STS) and 2 μL L⁻¹ 1-methylcyclopropene (1-MCP), ethylene action inhibitors, and 50 mM α-aminoisobutyric acid, an inhibitor of 1-aminocyclopropane-1-carboxylate (ACC) oxidase, did not delay flower senescence. However, treatment with 1 mM l-α-(2-aminoethoxyvinyl) glycine, an inhibitor of ACC synthase, slightly delayed flower senescence. Pollination significantly accelerated petal senescence of G. scabra flowers. Ethylene production of petals, gynoecium, and stamens in unpollinated flowers slightly increased during senescence. Pollination significantly increased ethylene production of petals, gynoecium and stamens 1 day after pollination. To clarify whether 1-MCP delays senescence of cut gentian inflorescences, cut G. scabra ‘Yuki-hotaru’, G. scabra × Gentiana triflora ‘Aoi-kaze’, and G. triflora ‘Koharu’ inflorescences with various stages of flowers, including buds with colored petals, were treated with 2 μL L⁻¹ 1-MCP for 24 h. 1-MCP treatment delayed flower wilting of cut inflorescences of ‘Aoi-kaze’ and ‘Yuki-hotaru’ more than that of ‘Koharu’, suggesting that there is species variation in the effect of 1-MCP in delaying flower senescence of cut gentian inflorescences.     

Imazalil residue loading and green mould control in citrus packhouses

Imazalil (IMZ) is commonly applied in South African citrus packhouses for the control of green mould, caused by Penicillium digitatum, yet the disease still causes significant postharvest losses. The maximum residue limit (MRL) for IMZ on citrus fruit is 5 μg g⁻¹, whereas 2-3 μg g⁻¹ is a biologically effective residue level that should at least inhibit green mould sporulation. Standard compliance auditing of residue levels of citrus fruit, however, indicate that fruit from the majority of packhouses have residues of ≈1 μg g⁻¹. Poor disease control from insufficient residue loading might further be compounded by the presence of IMZ-resistant isolates of P. digitatum in packhouses. This study was conducted to assess the current status of IMZ application in South African packhouses, to determine the adequate residue levels needed to control green mould and inhibit its sporulation using both IMZ sensitive and resistant isolates, to investigate IMZ application methods and resultant residue levels in commercial citrus packhouses, and to study optimisation of modes of IMZ application in citrus packhouses. Factors studied were IMZ concentration, application type (spray vs. dip and drench), exposure time, solution temperature and pH, as well as curative and protective control of P. digitatum. The packhouse survey showed that the majority of packhouses applied IMZ in a sulphate salt formulation through a fungicide dip tank, and loaded an IMZ residue of ≈1 μg g⁻¹. In dip applications, IMZ had excellent curative and protective activity against Penicillium isolates sensitive to IMZ. However, curative control of IMZ resistant isolates was substantially reduced and protective control was lost, even at twice the recommended concentration, nor was sporulation inhibited. The use of sodium bicarbonate (2%) buffered imazalil sulphate solutions at pH ±8, compared with pH ±3 of the unbuffered solutions, markedly increased IMZ residue loading on Navel and Valencia oranges and improved curative and protective control of IMZ resistant isolates. Exposure time did not affect IMZ residue loading in IMZ sulphate solutions at pH 3, although the MRL was exceeded after 45 s exposure in pH 8 solutions. Imazalil applied through spray or drench application improved residue loading, but green mould control was less effective than after dip application.     

Adaptability and variation in <Emphasis Type="Italic">Isatis tinctoria</Emphasis> L.: a new crop for Europe

Isatis tinctoria L. was cultivated until the 19th century to produce indigo, a natural blue pigment used principally for dyestuffs. The current search for alternative crops and interest in natural products has led to reconsidering I. tinctoria as a crop to be grown in marginal areas to produce natural indigo. To reintroduce I. tinctoria into cultivation, its behaviour under different climatic conditions as well as its morpho-physiological and genetic diversity must be assessed in order to evaluate the possibilities of future breeding work. To do this, a Eurasian collection of 15 accessions was studied in a 2-year experiment. The study was carried out in four locations in order to assess plant performance at altitudes ranging from 380 to 1,700 m a.s.l. A second experiment evaluated the morpho-physiological diversity of several traits (some related to agronomic performances) of the collection. In a third experiment the genetic traits of the collection were characterised by using eight AFLP and eight SAMPL markers. The species showed a wide adaptability to different mountainous conditions and the populations showed high morphologic and genetic variability and differed according to their origins. Both morpho-physiological and molecular characterisation allowed the accessions to be distinguished into groups of European and Asian origin. Future breeding work is recommended because some accessions have good agronomic potential.