The maturation profile triggers differential expression of sugar metabolism genes in melon fruits.

Melons are commercially important crops that requires specific quality attributes for successful commercialization, including accumulation of sugars, particularly sucrose. This trait can be influenced by various factors, such as the type of ripening. Cucumis melo L. is an ideal species for studying sugar metabolism because it has both climacteric and non-climacteric cultivars. Thus, this study aimed to examine the gene expression of sucrose metabolism candidates using RT-qPCR, in conjunction with postharvest physiological analyzes and high-performance liquid chromatography-based sugar quantification, in the melon cultivars 'Gaúcho' (climacteric) and 'Eldorado' (non-climacteric). The results showed that sucrose synthase 1 played a role in the synthesis and accumulation of sucrose in both cultivars, whereas sucrose synthase 2 was more highly expressed in 'Gaúcho', contributing to lower hexose content. Invertase inhibitor 1 was more highly expressed in 'Eldorado' and may be involved in sugar-induced maturation. Neutral α-galactosidase had distinct functions, playing a role in substrate synthesis for the growth of young 'Eldorado' fruits, whereas in mature 'Gaúcho' fruits it participated in the metabolism of raffinose family oligosaccharides for sucrose accumulation. The expression of trehalose-6-phosphate synthase genes indicated a greater involvement of these enzymes in the sugar regulation in 'Gaúcho' melons. These findings shed light on the intraspecific differences related to fruit quality attributes in different types of maturation and contribute to a deeper understanding of the underlying molecular mechanisms involved in the metabolism of sugars in melons, which can inform breeding programs aimed at improving fruit quality attributes in this crop.

Comparison of Volatile Compounds Contributing to Flavor of Wild Lowbush (Vaccinium augustifolium) and Cultivated Highbush (Vaccinium corymbosum) Blueberry Fruit Using Gas Chromatography-Olfactometry.

The flavor of blueberry fruit products is an important parameter determining consumer satisfaction. Wild lowbush blueberries are primarily processed into products, but their flavor chemistry has not been characterized. The objective of this study was to characterize the aroma chemistry of lowbush blueberries and compare it with that of highbush. Aroma volatiles of lowbush blueberries from four Canadian provinces and five highbush blueberry cultivars were isolated using headspace solid-phase microextraction (SPME) and characterized using gas chromatography-olfactometry (GC-O) and 2-dimensional gas chromatography-time of flight-mass spectrometry (GC×GC-TOF-MS). Lowbush fruit volatiles were composed of 48% esters, 29% aldehydes and 4% monterpenoids compared to 48% aldehydes, 26% monoterpenoids and 3% esters in highbush fruit. Twenty-three aroma-active peaks were identified in lowbush compared to forty-two in highbush fruit using GC-O. The most aroma-active compounds in lowbush fruit were ethyl 2-methylbutanoate, methyl 2-methylbutanoate, methyl 3-methylbutanoate, ethyl 3-methylbutanoate and ethyl propanoate compared to geraniol, (Z)-3-hexen-1-ol, 1-octen-3-one, α-terpineol and linalool in highbush fruit. The aroma volatile composition was more consistent among lowbush fruit samples than the five highbush cultivars. Aroma-active GC-O peaks were described more frequently as "floral", "fruity", "sweet" and "blueberry" in lowbush than in highbush fruit. Results suggest wild lowbush blueberries would provide "fruitier" and "sweeter" flavors to food products than cultivated highbush fruit.

Phytochemical Enhancement in Broccoli Florets after Harvest by Controlled Doses of Ozone.

The objective of this work was to examine the effect of controlled doses of O3 (0, 5 µL L-1 of O3 for 60 min, and 5 µL L-1 of O3 for 720 min) on the quality and phytochemical content of broccoli florets during postharvest storage. The optimal dose was found at 5 µL L-1 of O3 for 60 min, from the color retention of broccoli florets exposed to the gas treatment. Overall, the antioxidant capacity of the florets was significantly affected by both doses of O3 compared to the non-exposed florets. The profile of glucosinolates was determined for up to 14 days in broccoli florets stored at 4 °C by LC-MS. The amount of total glucobrassicins and total hydroxy-cinnamates in florets significantly (p ≤ 0.05) improved by the application of 5 µL L-1 of O3 for 60 min compared to non-treated florets. The up-regulation of genes of the tryptophan-derived glucosinolate pathway was observed immediately after both treatments. The gene expression of CYP79A2 and CYP79B3 in broccoli was significantly higher in broccoli florets exposed to 5 µL L-1 of O3 for 720 min compared to non-exposed florets. Although enhancement of secondary metabolites can be achieved by the fumigation of broccoli florets with low doses of ozone, quality parameters, particularly weight loss, can be compromised.

Apple Ripening Is Controlled by a NAC Transcription Factor.

Softening is a hallmark of ripening in fleshy fruits, and has both desirable and undesirable implications for texture and postharvest stability. Accordingly, the timing and extent of pre-harvest ripening and associated textural changes following harvest are key targets for improving fruit quality through breeding. Previously, we identified a large effect locus associated with harvest date and firmness in apple (Malus domestica) using genome-wide association studies (GWAS). Here, we present additional evidence that polymorphisms in or around a transcription factor gene, NAC18.1, may cause variation in these traits. First, we confirmed our previous findings with new phenotype and genotype data from ∼800 apple accessions. In this population, we compared a genetic marker within NAC18.1 to markers targeting three other firmness-related genes currently used by breeders (ACS1, ACO1, and PG1), and found that the NAC18.1 marker was the strongest predictor of both firmness at harvest and firmness after 3 months of cold storage. By sequencing NAC18.1 across 18 accessions, we revealed two predominant haplotypes containing the single nucleotide polymorphism (SNP) previously identified using GWAS, as well as dozens of additional SNPs and indels in both the coding and promoter sequences. NAC18.1 encodes a protein that is orthogolous to the NON-RIPENING (NOR) transcription factor, a regulator of ripening in tomato (Solanum lycopersicum). We introduced both NAC18.1 transgene haplotypes into the tomato nor mutant and showed that both haplotypes complement the nor ripening deficiency. Taken together, these results indicate that polymorphisms in NAC18.1 may underlie substantial variation in apple firmness through modulation of a conserved ripening program.

Metabolic Profile of Strawberry Fruit Ripened on the Plant Following Treatment With an Ethylene Elicitor or Inhibitor.

Strawberry is the most studied nonclimacteric fruit for understanding the role ethylene has in ripening regulation. However, previous studies on the effects of ethylene on strawberry ripening were conducted with detached fruit. Thus, the aim of this work was to determine the effect of ethylene and the ethylene-action inhibitor 1-methylcyclopropene (1-MCP) applied at different developmental stages on important physical-chemical attributes of ripe 'Albion' strawberry. Fruit at four developmental stages that remained attached to the plant were dipped in one of three treatment solutions (Ethephon, 1-methylcyclopropene, and water), plus one absolute control that received no dip. Following treatment, when immature fruit were fully red or 24 h after treatment for red-treated fruit, strawberry fruit were assessed for physicochemical properties (mass, length, diameter, firmness, color, titratable acidity, soluble solids, pH, total phenolics, sugar, organic acid, amino acid, and volatile composition). The days following treatment required for fruit to ripen were also recorded. Treatments did not affect the rate of ripening nor fruit color, titratable acidity, pH, soluble solids, total phenolics, sugars, or organic acids of ripe fruit. Ethephon affected fruit mass, diameter, length, firmness, anthocyanins, amino acids, and volatiles, but these effects were dependent on fruit developmental stage at which the treatment was applied. When green fruit were treated with ethephon, ripe fruit had larger diameter and mass. Ethephon treatment of white fruit resulted in ripe fruit having greater anthocyanin content. Treatment of pink fruit resulted in ripe fruit having smaller diameter, length, and mass and greater firmness. Treatment of red fruit with ethephon altered fruit volatile composition, increasing concentrations of ethyl- and acetate-esters, which were reduced by the 1-MCP treatment. Ethephon treatment increased concentrations of 11 of the 19 free amino acids measured in ripe fruit with treatment of green and white fruit having the greatest effect. A total of 41 volatile compounds had significant correlations with 14 amino acids. While ethylene did not stimulate typical ripening of strawberry fruit, it does appear to alter fruit development and metabolism. The physiological effects of ethylene on strawberry fruit appear to depend on the developmental stage of the fruit.

Genome-wide association studies in apple reveal loci of large effect controlling apple polyphenols.

Apples are a nutritious food source with significant amounts of polyphenols that contribute to human health and wellbeing, primarily as dietary antioxidants. Although numerous pre- and post-harvest factors can affect the composition of polyphenols in apples, genetics is presumed to play a major role because polyphenol concentration varies dramatically among apple cultivars. Here we investigated the genetic architecture of apple polyphenols by combining high performance liquid chromatography (HPLC) data with ~100,000 single nucleotide polymorphisms (SNPs) from two diverse apple populations. We found that polyphenols can vary in concentration by up to two orders of magnitude across cultivars, and that this dramatic variation was often predictable using genetic markers and frequently controlled by a small number of large effect genetic loci. Using GWAS, we identified candidate genes for the production of quercitrin, epicatechin, catechin, chlorogenic acid, 4-O-caffeoylquinic acid and procyanidins B1, B2, and C1. Our observation that a relatively simple genetic architecture underlies the dramatic variation of key polyphenols in apples suggests that breeders may be able to improve the nutritional value of apples through marker-assisted breeding or gene editing.

Coencapsulation of Polyphenols and Anthocyanins from Blueberry Pomace by Double Emulsion Stabilized by Whey Proteins: Effect of Homogenization Parameters.

Blueberry pomace is a rich source of high-value bioactive polyphenols with presumed health benefits. Their incorporation into functional foods and health-related products benefits from coencapsulation and protection of polyphenol-rich extracts in suitable carriers. This study aimed to create a water-in-oil-in-water (W1/O/W2) double emulsion system suitable for the coencapsulation of total phenolics (TP) and anthocyanins (TA) from a polyphenol-rich extract of blueberry pomace (W1). The effect of critical physical parameters for preparing stable double emulsions, namely homogenization pressure, stirring speed and time, was investigated by measuring the hydrodynamic diameter, size dispersity and zeta potential of the oil droplets, and the encapsulation efficiency of TP and TA. The oil droplets were negatively charged (negative zeta potential values), which was related to the pH and composition of W2 (whey protein isolate solution) and suggests stabilization by the charged whey proteins. Increasing W1/O/W2 microfluidization pressure from 50 to 200 MPa or homogenization speed from 6000 to 12,000 rpm significantly increased droplet diameter and zeta potential and decreased TA and TP encapsulation efficiency. Increasing W1/O/W2 homogenization time from 15 to 20 min also increased droplet diameter and zeta potential and lowered TA encapsulation efficiency, while TP encapsulation did not vary significantly. In contrast, increasing W1/O homogenization time from 5 to 10 min at 10,000 rpm markedly increased TA encapsulation efficiency and reduced droplet diameter and zeta potential. High coencapsulation rates of blueberry polyphenols and anthocyanins around 80% or greater were achieved when the oil droplets were relatively small (mean diameter < 400 nm), with low dispersity (<0.25) and a high negative surface charge (-40 mV or less). These characteristics were obtained by homogenizing for 10 min at 10,000 rpm (W1/O), then 6000 rpm for 15 min, followed by microfluidization at 50 MPa.

Influence of Extraction Conditions on Ultrasound-Assisted Recovery of Bioactive Phenolics from Blueberry Pomace and Their Antioxidant Activity.

The increase in diet-related chronic diseases has prompted the search for health-promoting compounds and methods to ensure their quality. Blueberry pomace is a rich yet underutilized source of bioactive polyphenols. For these high-value bioactive molecules, ultrasound-assisted extraction (USAE) is an attractive and green alternative to conventional extraction techniques for improving purity and yields. This study aimed to assess the impact of USAE parameters (sonication time, solvent composition, solid/liquid ratio, pH and temperature) on the recovery of phenolic compounds from blueberry pomace and antioxidant activity of the extracts. Total phenolic, flavonoid and anthocyanin contents (TPC, TFC and TAC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity were analysed. USAE in 50% ethanol/water was the most efficient, yielding the highest TPC (22.33 mg/g dry matter (DM)), TFC (19.41 mg/g DM), TAC (31.32 mg/g DM) and DPPH radical scavenging activity (41.79 mg Trolox/g DM). USAE in water showed the lowest values even at low (1/40) solid/liquid ratio (7.85 mg/g DM, 3.49 mg/g DM, and 18.96 mg/g DM for TPC, TFC and TAC, respectively). Decreasing the solid/liquid ratio in water or 50% ethanol significantly increased TPC, TFC, TAC and DPPH radical scavenging. With ethanol, increasing the temperature in the range 20⁻40 °C decreased TPC but increased TFC and DPPH radical scavenging activity. Anthocyanin profiles of water and ethanolic extracts were qualitatively similar, consisting of malvidin, delphinidin, petunidin and cyanidin. These findings indicate that USAE is a method of choice for extracting high-value bioactive phenolics from blueberry pomace. Selective enrichment of different phenolic fractions is possible under select extraction conditions.

Impact of Listeria Inoculation and Aerated Steam Sanitization on Volatile Emissions of Whole Fresh Cantaloupes.

Rapid methods to detect bacterial pathogens on food and strategies to control them are needed to mitigate consumer risk. This study assessed volatile emissions from whole cantaloupe melons (Cucumis melo) as an indicator of Listeria contamination and in response to steam vapor decontamination. Cantaloupe were inoculated with Listeria innocua, a nonpathogenic surrogate for L. monocytogenes, then exposed to 85 °C steam for 240 s (4 min) followed by rapid chilling and storage for 0, 7, 10, or 14 days at 4, 7, or 10 °C. Volatile emissions from whole melons were collected on Carbopack B/Carboxen 1000 headspace collection tubes and analyzed by gas chromatography-mass spectroscopy following thermal desorption. Introduction of L. innocua to cantaloupe rind resulted in a reduction of aromatic compound emission. However, this response was not unique to Listeria contamination in that steam vapor treatment also reduced emission of these compounds. As well, steam vapor treatment diminished the number of viable Listeria and indigenous microflora while causing physiological injury to melon rind. Heat treatment had no significant effects on flesh firmness, color, titratable acidity, or soluble solids, but the production of typical aroma volatiles during postharvest ripening was inhibited. No unique volatile compounds were detected in Listeria contaminated melons. While changes in volatile emissions were associated with Listeria inoculation, they could not be differentiated from heat treatment effects. Results indicate that volatile emissions cannot be used as a diagnostic tool to identify Listeria contamination in whole cantaloupe melons. PRACTICAL APPLICATION: The detection of pathogen contamination on fresh produce is a continuing challenge. Using a nondestructive screening method, the presence of surrogate Listeria innocua on fresh whole cantaloupes was shown to alter the emissions of aromatic volatiles from whole cantaloupes. However, these altered emissions were not found to be unique to Listeria spp. and therefore cannot be used as a definitive indicator of Listeria contamination.

Aerated Steam Sanitization of Whole Fresh Cantaloupes Reduces and Controls Rind-Associated Listeria but Enhances Fruit Susceptibility to Secondary Colonization.

Recent bacterial illnesses and outbreaks associated with the consumption of fresh and fresh-cut fruit and vegetables emphasize the need to supply produce that is microbiologically safe while retaining its quality and nutrient value. We assessed the capacity of aerated steam to reduce initial levels and control the posttreatment proliferation of a 4-strain mixture of Listeria innocua, a surrogate for L. monocytogenes, and microflora native to the rind of whole cantaloupes. Studies were conducted at the pilot-scale level by passing deliberately contaminated melons through a prototype stainless-steel, continuous-feed heating device. Exposure for 240 s to aerated steam heated to 85 °C achieved a mean reduction in surface-inoculated L. innocua of 3.9 ± 0.6 log10 CFU/cm2 (n = 3) and decreased background microorganisms (yeast, moulds, and coliforms) to undetectable levels. No significant outgrowth of surviving L. innocua or yeast and moulds was observed on heat-treated melons during their storage at 4, 7, and 10 °C for 14 days. Treated fruit continued to respire. Although rind quality was altered, edible fleshy portions remained largely unaffected. Cantaloupe inoculated with L. innocua subsequent to its exposure to aerated steam provided a suitable environment for surrogate growth (mean 3.3 log10 increase in rind density over 10 days at 7 °C), whereas its proliferation was restricted on nonheated cantaloupe (mean 0.7 log10 increase). Steam sanitization provides an effective means for the control of pathogen and spoilage organisms, but the proliferation of surrogate organisms on heated cantaloupes raises concern regarding the impact of postprocessing contamination on consumer health risk. PRACTICAL APPLICATION: Water vapor (steam) at a high temperature can be used to sanitize the surface of fresh, whole cantaloupe melons in a continuous-feed manner. Both Listeria bacteria and spoilage organisms are markedly reduced from initial levels and survivor outgrowth severely restricted during subsequent refrigerated storage. This approach to microorganism control is likely most applicable in situations where rinds and flesh are to be separated immediately via further processing.

A Genome-Wide Association Study of Apple Quality and Scab Resistance.

The apple ( × Borkh.) is an economically and culturally important crop grown worldwide. Growers of this long-lived perennial must produce fruit of adequate quality while also combatting abiotic and biotic stress. Traditional apple breeding can take up to 20 yr from initial cross to commercial release, but genomics-assisted breeding can help accelerate this process. To advance genomics-assisted breeding in apple, we performed genome-wide association studies (GWAS) and genomic prediction in a collection of 172 apple accessions by linking over 55,000 single nucleotide polymorphisms (SNPs) with 10 phenotypes collected over 2 yr. Genome-wide association studies revealed several known loci for skin color, harvest date and firmness at harvest. Several significant GWAS associations were detected for resistance to a major fungal pathogen, apple scab ( [Cke.] Wint.), but we demonstrate that these hits likely represent a single ancestral source. Using genomic prediction, we show that most phenotypes are sufficiently predictable using genome-wide SNPs to be candidates for genomic selection. Finally, we detect a signal for firmness retention after storage on chromosome 10 and show that it may not stem from variation in , a gene repeatedly identified in bi-parental mapping studies and widely believed to underlie a major QTL for firmness on chromosome 10. We provide evidence that this major QTL is more likely due to variation in a neighboring ethylene response factor (ERF) gene. The present study showcases the superior mapping resolution of GWAS compared to bi-parental linkage mapping by identifying a novel candidate gene underlying a well-studied, major QTL involved in apple firmness.

QTL analysis of soft scald in two apple populations.

The apple (Malus×domestica Borkh.) is one of the world's most widely grown and valuable fruit crops. With demand for apples year round, storability has emerged as an important consideration for apple breeding programs. Soft scald is a cold storage-related disorder that results in sunken, darkened tissue on the fruit surface. Apple breeders are keen to generate new cultivars that do not suffer from soft scald and can thus be marketed year round. Traditional breeding approaches are protracted and labor intensive, and therefore marker-assisted selection (MAS) is a valuable tool for breeders. To advance MAS for storage disorders in apple, we used genotyping-by-sequencing (GBS) to generate high-density genetic maps in two F1 apple populations, which were then used for quantitative trait locus (QTL) mapping of soft scald. In total, 900 million DNA sequence reads were generated, but after several data filtering steps, only 2% of reads were ultimately used to create two genetic maps that included 1918 and 2818 single-nucleotide polymorphisms. Two QTL associated with soft scald were identified in one of the bi-parental populations originating from parent 11W-12-11, an advanced breeding line. This study demonstrates the utility of next-generation DNA sequencing technologies for QTL mapping in F1 populations, and provides a basis for the advancement of MAS to improve storability of apples.

Targeted quantitative proteomic investigation employing multiple reaction monitoring on quantitative changes in proteins that regulate volatile biosynthesis of strawberry fruit at different ripening stages.

A targeted quantitative proteomic investigation employing the multiple reaction monitoring (MRM, SRM) technique was conducted on strawberry fruit at different development stages. We investigated 22 proteins and isoforms from 32 peptides with 111 peptide transitions, which may be involved in the volatile aroma biosynthesis pathway. The normalized protein abundance was significantly changed in coincidence with increased volatile production and advanced fruit maturities. Among them, alcohol acyltransferase (AAT), quinone oxidoreductase (QR), malonyl Co-A decarboxylase, (MLYCD), pyruvate decarboxylase (PDC), acetyl Co-A carboxylase (ACCase), and acyl Co-A synthetase (ACAs) were increased significantly. Several alcohol dehydrogenases (ADHs), and 3-oxoacyl-ACP synthase were significantly decreased. Furthermore, the expression of seven genes related to strawberry volatile production was also investigated using real-time qPCR. Among the tested genes, QR, AAT, ACCase, OMT, PDC and ADH showed increased up-regulation during fruit ripening, while 3-isopropylmalate dehydrogenase (IMD) decreased. Strong correlation between quantitative proteomic data and gene expression suggested that AAT, QR, ACCase, and PDC played critical roles in volatile biosynthesis of strawberry during fruit ripening. Poor correlation between protein abundance and gene expression of ADH was found.

A method to detect diphenylamine contamination of apple fruit and storages using headspace solid phase micro-extraction and gas chromatography/mass spectroscopy.

Analysis of headspace concentrations of diphenylamine using solid phase micro-extraction (SPME) was examined for its suitability to detect DPA contamination and off-gassing in apple (Malus domestica) fruit, storage rooms and storage materials. Four SPME fibre coatings including polydimethylsiloxane (PDMS, 100 µm), PDMS/divinylbenzene (PDMS/DVB), Polyacrylate (PA) and PDMS 7 µm were evaluated. The average limits of detection and of quantification for head space DPA ranged from 0.13 to 0.72 µg L(-1) and 0.42 to 2.35 µg L(-1), respectively. Polyacrylate was identified to be the most suitable and compatible fibre for DPA analysis in apple samples, because of its high sensitivity to DPA and low fruit volatile interferences. SPME techniques were further applied to study contamination of DPA in apples, storage rooms and packaging materials. DPA was found in the air of storage rooms containing apples that were not treated with DPA. Wood and plastic bin material, bin liners, and foam insulation all adsorbed and off-gassed DPA and could be potential sources of contamination of untreated apples.

Effect of ozone pre-conditioning on quality and antioxidant capacity of papaya fruit during ambient storage.

The objective of this study was to compare the physico-chemical characteristics and antioxidant activity of ozone-treated papaya fruit and untreated fruit. Freshly harvested papaya fruit were exposed continuously to ozone fumigation (0, 1.5, 2.5, 3.5 and 5ppm) for 96h prior to ambient storage at 25±3°C and 70±5% relative humidity (RH) for up to 14days. The fruit exposed to 2.5ppm ozone had higher levels of total soluble solids (25.0%), ascorbic acid content (12.4%), ß-carotene content (19.6%), lycopene content (52.1%), and antioxidant activity (30.9%), and also reduced weight loss (11.5%) at day 10 compared to the control. The sensory attributes of papaya treated with 2.5ppm ozone was superior in sweetness and overall acceptability. These results support the application of ozone as a non-thermal and safe food preservation technique for papaya which can benefit both the producers and consumers.

Characterization of Changes in Polyphenols, Antioxidant Capacity and Physico-Chemical Parameters during Lowbush Blueberry Fruit Ripening.

Changes in major polyphenols, antioxidant capacity, and selected physico-chemical parameters were examined in lowbush blueberry during fruit ripening. Polyphenols (phenolic acids, flavonols, flavan-3-ols, and anthocyanins), density, soluble solid content, pH, titratable acidity, sugars, organic acids, and antioxidant capacity were determined in fruits of four maturities: green, pink/red, blue, and over-mature. Highest concentrations of flavonols, flavan-3-ols, and phenolic acids were in green fruits: 168 ± 107, 119 ± 29 and 543 ± 91 mg/100 g dry weight (DW) respectively. Highest anthocyanin levels were found in blue and over-mature fruits (1011-1060 mg/100 DW). Chlorogenic acid was the most abundant phenolic acid and quercetin-3-O-galactoside the most abundant flavonol in all maturities. Epicatechin was the most abundant flavan-3-ol in green fruits (80 ± 20 mg/100 DW), and catechin was the most abundant in other maturity stages. Increase of glucose and fructose and decrease of organic acids were observed during fruit ripening. Among six organic acids found, quinic acid (1.7-9.5 mg/100 mg DW) was the most abundant throughout the fruit ontogeny. Soluble solids, pH, and density increased with maturity while, titratable acidity decreased. These findings can be helpful in optimizing harvest and processing operations in lowbush blueberry fruits.

Effect of hexanal vapor on the growth of postharvest pathogens and fruit decay.

The effect of the natural volatile hexanal was studied as an antifungal agent on the major postharvest fungal pathogens Botrytis cinerea, Monilinia fructicola, Sclerotinia sclerotiorum, Alternaria alternata, and Colletotrichum gloeosporioides. The antifungal effect of hexanal vapor was dependent on concentration and treatment duration, but sensitivity of the pathogens varied. All spores of B. cinerea and M. fructicola were killed after exposure to 900 microL/L for 12 h at 20 degrees C, and almost all were killed after a 24-h exposure to 450 microL/L. Only moderate numbers of spores were killed at a concentration of 200 microL/L. Mycelial growth of S. sclerotiorum on agar was completely inhibited after a 12-h exposure to 900 microL/L, but only slight inhibition occurred at 450 microL/L and none at 200 microL/L. Mycelium of A. alternata and C. gloeosporioides appeared more sensitive, with strong inhibition occurring after a 12-h exposure at 450 microL/L. Similar trends in spore viability and mycelial growth were observed at 7 degrees C. The antifungal effect of hexanal vapor was further tested on raspberry fruit naturally infected with B. cinerea and on peach fruit inoculated with spores of M. fructicola. Decay was markedly reduced in raspberry and almost completely controlled in peach after exposure to 900 microL/L hexanal vapor for 24 h. The potential of hexanal for postharvest decay control is discussed.

A rapid capillary gel electrophoresis method for the quantitative determination of RuBisCo in spinach.

A capillary gel electrophoretic (CGE) method for the quantitative analysis of RuBisCo in spinach leaves was developed. RuBisCo was resolved into large and small subunits in the presence of sodium dodecyl sulphate (SDS) by the CGE procedure which enabled the determination of the molecular weight of each unit accurately; the values so determined were in close agreement with those reported using other methods. Advantages of CGE over SDS-polyacrylamide gel electrophoresis and high-pressure gel filtration include decreased sample preparation and analysis time, superior resolution and greater sensitivity permitting reduced sample size and trace analysis. In addition, CGE provided precise quantification of RuBisCo and was demonstrated to be a viable alternative to other available methods of protein analysis.

Using a modified ferrous oxidation-xylenol orange (FOX) assay for detection of lipid hydroperoxides in plant tissue.

The ferrous oxidation-xylenol orange (FOX) assay was adapted for quantifying lipid hydroperoxides (LOOHs) in plant extracts. Excised pieces of several fruit and vegetable species were exposed to 83 kJ m(-2) day(-1) of biologically effective ultraviolet B irradiance (UV-B(BE)) for 10-12 days to induce cellular oxidation. The LOOH and thiobarbituric acid reactive substance (TBARS) concentrations of these plant tissues were assessed with the FOX and iodometric assays for the former and a modified TBARS assay for the latter. There was generally good agreement between the FOX and iodometric methods both prior to and following the UV exposure. However, the iodometric assay appeared to have some difficulty in consistently quantifying lower LOOH levels (<11 microM), whereas the FOX assay measured LOOH concentrations as low as 5 microM. All tissues exhibited UV-induced increases in TBARS, indicating a marked degree of cellular oxidation in the exposed tissue segments. Compared with the iodometric assay, the FOX method consistently generated less variable LOOH values. The presence of authentic linoleic acid-OOHs in spiked avocado and spinach samples (11 microM) was identified with liquid chromatography-mass spectrometry techniques, which validated corresponding FOX assay results. The FOX method is inexpensive, is not sensitive to ambient O2 or light levels, and can rapidly generate LOOH measurements. The physiological value of the FOX assay resides in its ability to measure initial rather than more advanced fatty acid oxidation; hence, early membrane-associated stress events in plant tissue can be detected.