Tracking the development of the superficial scald disorder and effects of treatments with diphenylamine and 1-MCP using an untargeted metabolomic approach in apple fruit.

Superficial scald is a physiological storage disorder that significantly reduces the marketability of apple fruit. To gain fundamental knowledge about the biochemical pathways leading to the development of the disorder and mechanisms of treatments for prevention, an untargeted metabolomics experiment employing liquid chromatography and mass spectrometry with data independent acquisition was performed. Metabolomic changes of two apple cultivars 'Cortland' and 'Red Delicious' with scald development and scald control treatments, using diphenylamine and 1-MCP, at 0-1 °C for up to 7 months was investigated. In total, 833 features/compounds were analyzed, and among them 59 were found to change significantly in controls involved in scald development, and in response to DPA and 1-MCP treatments. Our results provide new evidence that metabolites in association with phenylpropanoid metabolism, antioxidant and redox systems, and amino acid metabolism are related closely to scald development and response to potential treatments.

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.

Optimized extraction and characterization of antimicrobial phenolic compounds from mangosteen (Garcinia mangostana L.) cultivation and processing waste.

BACKGROUND: Applications for antimicrobials derived from the mangosteen (Garcinia mangostana L.) plant are presently restricted by high production costs. Extraction from cultivation or processing waste streams using a solvent-free approach could lessen to permit commercial applications in food processing and preservation. RESULTS: Phenolics were extracted from mangosteen bark, leaf and fruit pericarp in methanol and in water using response surface methodology to optimize recovery. Initial examination of antimicrobial effects revealed a lack of antimicrobial activity against fungi and weak activity against the Gram-negative bacteria Escherichia coli and Salmonella typhimurium. In contrast, extracts prepared from bark or fruit pericarp exhibited strong pH-dependent bacteriostatic and bactericidal effects against Listeria monocytogenes and Staphylococcus aureus. Activity was slightly weaker in aqueous extracts due to lower concentrations of tartaric acid esters and flavonols than in methanolic extracts. Measurement of propidium iodide uptake and ATP leakage indicated that the extracts induced damage to the membrane of Gram-positive bacteria. CONCLUSION: Extracts of mangosteen bark and fruit pericarp contain mixtures of phenolic compounds with activity against Gram-positive bacteria, notably Listeria monocytogenes. Extraction of phenolics from mangosteen waste could yield fractions for potential applications in the formulation of low-cost processing aids or sanitizers for the food industry.

Effects of 1-methylcyclopropene and post-controlled atmosphere air storage treatments on fresh-cut Ambrosia apple slices.

BACKGROUND: The effect of 1-methylcyclopropene (1-MCP) treatment and two different post-controlled atmosphere air storage (PCAAS) durations on the quality and chemistry of fresh-cut Ambrosia apple slices was studied. RESULTS: PCAAS for 1 or 2 weeks prior to slicing had an overall positive effect on the resultant quality of fresh-cut apple slices. The most significant responses to PCAAS were the suppression of both phenolic and o-quinone accumulation in slices, and this was related to the significantly lower browning potential values obtained for slices from PCAAS-treated apples. Polyphenol oxidase (PPO), peroxidase (POX) and ascorbate peroxidase (APOX) activities were not affected by 1-MCP or PCAAS treatments. PPO and POX activities were almost completely inhibited by a 50 g L⁻¹ calcium ascorbate anti-browning dip of apple slices from all treatments. CONCLUSION: The most dramatic effect of the PCAAS treatments was to reduce the accumulation of soluble phenolics, which is likely the reason that o-quinone accumulation was also inhibited in treated fruits. The consequent reduction in browning potential may be the explanation as to why PCAAS treatment has been shown to reduce fresh apple slice browning in previous work.

Modulation of wound-induced hydrogen peroxide and its influence on the fate of Escherichia coli O157:H7 in cut lettuce tissues.

Wounding of lettuce tissue has been examined previously by others in regard to browning reactions, and treatments to modulate wounding responses were evaluated for reduction of browning. However, the wounding process also releases oxygen radicals such as hydrogen peroxide. This study focused on the evaluation of two treatments that reduce hydrogen peroxide at cut surfaces (heat treatment and pyruvate addition) and one treatment that enhances its production (infusion with the fungal elicitor harpin). Hydrogen peroxide changes in response to treatment were also associated with resultant survival of Escherichia coli O157:H7, which was inoculated onto the lettuce before cutting. Heat-treated lettuce produced significantly less hydrogen peroxide, and microbial analysis showed that E. coli O157:H7 survival on packaged, heat-treated lettuce was higher than on non-heat-treated controls. Lettuce was also cut under a solution of sodium pyruvate (a well-known hydrogen peroxide quencher), and E. coli O157:H7 survival was found to be enhanced with that treatment. When lettuce was infused with harpin before cutting, hydrogen peroxide production was enhanced, and this was associated with reduced survival of E. coli O157:H7. These results collectively support the hypothesis that modulation of wound-generated hydrogen peroxide can have an influence on E. coli O157:H7 survival on cut and packaged romaine lettuce.

Slicing under chlorinated water improves the disinfection of fresh-cut romaine lettuce.

The aim of this study was to determine whether the timing or mode of application influences the antimicrobial efficacy of a sodium hypochlorite solution against microorganisms on fresh-cut Romaine lettuce. Lettuce leaves were treated with a solution of sodium hypochlorite containing 70 mg liter(-1) free chlorine prior to slicing (WLS), while submerged in the solution (CWS), or after slicing (PCS), the last being common practice in the fresh-cut industry. Immediate antimicrobial effects were determined from analyses performed directly after treatment, while delayed effects were derived from analysis of packaged samples stored at 4 and 15°C. Experiments were performed with both uninoculated and inoculated whole leaves, using Escherichia coli O157:H7 from a type collection as an inoculum. The CWS treatment provided the greatest initial reductions in aerobic plate counts of E. coli O157:H7 populations, followed by PCS and WLS. The advantage afforded by the CWS approach was maintained over 10 days in storage at 4°C. In addition, it was noted that the visual quality attributes of the fresh-cut lettuce from the CWS approach were better than those from the other two treatments. These results collectively suggest that slicing lettuce under a sanitizer solution could provide better disinfection of fresh-cut Romaine lettuce than the current practice of washing with sanitizer after cutting has taken place.

Effect of short-term air storage after removal from controlled-atmosphere storage on apple and fresh-cut apple quality.

BACKGROUND: One of the realities of apple distribution for long-term stored fruit is that a controlled-atmosphere (CA) storage room will be unsealed and fruit held in air storage and marketed over several weeks. This work was conducted to determine the effect of post-CA air storage of whole fruit on potential shelf life for fresh-cut apple slices. RESULTS: Fresh-cut slices of 'Spartan' and 'Delicious' apples held in post-CA air storage for 2 or 4 weeks showed the least changes in cut surface color as compared with those made from apples immediately on removal from CA. Shelf life was most improved by post-CA air storage in the 'Spartan' apples, which were more advanced in maturity as compared with the 'Delicious' apples. Internal ethylene concentration, firmness, and respiration changed significantly with post-CA air storage, suggesting a relationship between physiological status of the whole fruit and shelf life of slices made from that fruit. CONCLUSION: The results support the hypothesis that apples had suppressed physiological activity in CA storage and are susceptible to accelerated deterioration upon cutting. Holding fruit for 2 weeks in air storage allowed recovery of physiological activity, which resulted in greater resistance to deterioration in response to fresh-cut processing.

Effect of vanillin on the fate of Listeria monocytogenes and Escherichia coli O157:H7 in a model apple juice medium and in apple juice.

The effects of vanillin on the fates of Listeria monocytogenes and Escherichia coli O157:H7 at pH values between 3.5 and 4.5 were verified in a model apple juice (MAJ) medium and in apple juice incubated at 4 or 15 degrees C. Viable E. coli O157:H7 cells were recovered from MAJ for up to 10 days, but L. monocytogenes did not survive at pH 3.5. Supplementation with 40 mm vanillin exerted a lethal effect that was species, concentration, pH and temperature dependant. E. coli O157:H7 was more sensitive to vanillin than L. monocytogenes, and viable cells could not be recovered after 2 days incubation at either temperature. L. monocytogenes and E. coli O157:H7 were inoculated (10(5) cfu/ml) in pH adjusted (pH 4.00) or unadjusted (pH 3.42) juice from Granny Smith apples that was supplemented with 40 mm vanillin. Neither species were recovered after 3 days incubation at 4 or 15 degrees C. These findings indicate that vanillin could be useful as a preservative for minimally processed apple products.

Antimicrobial activity of vanillin against spoilage microorganisms in stored fresh-cut mangoes.

The antimicrobial activity of vanillin against four bacteria (Pantoea agglomerans, Aeromonas enteropelogenes, Micrococcus lylae, and Sphingobacterium spiritovorun), four fungi (Alternaria sp., Aspergillus sp., Penicillium sp., and Fusarium sp.), and three unidentified yeasts isolated from spoiling fresh-cut mango slices was verified in laboratory media adjusted to pH 5.0. MICs of vanillin against the fungi (12.5 to 13.3 mM), bacteria (10 to 13.3 mM), and yeasts (5.0 to 6.7 mM) indicated that all the test species were sensitive to the antimicrobial effects of vanillin. Fresh-cut mango slices were dipped for 1 min in solutions containing 40 and 80 mM vanillin before being packaged in rigid trays and stored at 5 and 10 degrees C to verify the effects of vanillin on the development of the spoilage microflora. Microbiological analysis for up to 14 days of storage revealed that treatment with 80 mM vanillin significantly delayed (P < 0.05) the development of total aerobic bacteria and yeast and mold populations. Vanillin may be a practical preservative for processing fresh-cut mango.

Destruction of Escherichia coli O157:H7 by vanillic acid in unpasteurized juice from six apple cultivars.

The behavior of Escherichia coli O157:H7 in Granny Smith, Gala, Empire, McIntosh, Red Delicious, and Golden Delicious apple juice with or without supplementation with 5 or 10 mM vanillic acid was examined over a storage period of 7 days at 4 and 15 degrees C. The consequences of supplementation on sensory difference and preference were also determined by triangle testing. Juices made from the six apple cultivars had pH values ranging between pH 3.13 and 3.92. Vanillic acid exerted a concentration, pH, and time-dependent lethal effect toward E. coli O157:H7 in unpasteurized apple juice. Supplementation with 10 mM vanillic acid led to a 5-logarithm reduction in populations after 7 days at both temperatures, but sensory analysis revealed significant differences from and preference for unsupplemented juices. Supplementation with 5 mM vanillic acid accelerated death of E. coli O157:H7, but population reductions ranged from 5 log CFU/ml in low pH juices to none in high pH juices, particularly at 4 degrees C. No sensory difference or preference was detected in two of the six juices at this level of supplementation.

Effect of pH on the inhibition of Listeria spp. by vanillin and vanillic acid.

The antimicrobial effects of vanillin and vanillic acid were verified against several species and strains of Listeria monocytogenes, Listeria innocua, Listeria grayi, and Listeria seeligeri in a laboratory medium adjusted to pH values ranging from 5.0 to 8.0. Medium pH had little influence on the MIC of vanillin as determined by a broth dilution assay, and growth of all test strains was inhibited by concentrations ranging from 23 to 33 mM. In contrast, none of the strains were inhibited by 100 mM vanillic acid at pH > 6.0, but complete inhibition was achieved at pH 5.0 with 10 mM. The effect of pH was further characterized by incubation of L. monocytogenes, L. innocua, and L. grayi in media containing 30 mM vanillin or 60 mM vanillic acid at pH 5.0, 6.0, and 7.0. Bactericidal effects increased with pH in media supplemented with vanillin. An inverse relationship was found for vanillic acid, and the lethality of the compound increased with declining pH. Mixtures of vanillin and vanillic acid exhibited additive inhibitory effects, particularly at lower pH. These natural antimicrobial compounds could prove useful either alone or in mixtures for the control of Listeria spp. in food products.

Warm water treatment in combination with modified atmosphere packaging reduces undesirable effects of irradiation on the quality of fresh-cut iceberg lettuce.

Fresh-cut iceberg lettuce dipped in either 5 or 47 degrees C water for 2 min was packaged in modified atmosphere film bags and then exposed to 0, 0.5, 1, or 2 kGy gamma-radiation. Dipping cut lettuce in 47 degrees C water for 2 min prior to irradiation reduced antioxidant and phenolic accumulations induced by irradiation. Irradiation at 2 kGy increased cellular leakage and sogginess of cut lettuce dipped in both temperatures. Samples irradiated at 0.5 and 1 kGy had similar firmness and vitamin C and antioxidant contents as the controls after 14 and 21 days of storage except 1 kGy samples dipped at 47 degrees C had lower antioxidant contents than controls at 14 days of storage. Lettuce dipped at 47 degrees C and irradiated at 0.5 and 1 kGy had better overall visual quality and less tissue browning than corresponding irradiated samples dipped at 5 degrees C. These results suggest lettuce treated with warm water and irradiated at 0.5 or 1 kGy had the best sensory quality without significant loss in texture, vitamin C, or total antioxidants.

Effect of wash water temperature and chlorination on phenolic metabolism and browning of stored iceberg lettuce photosynthetic and vascular tissues.

Cut tissues from distinct anatomical locations in iceberg lettuce (Lactuca sativa L.) were subjected to washing in cold (4 degrees C) and warm (47 degrees C) water with or without chlorine to assess their propensity to discoloration during storage. Total protein (Bradford method) and phenolic (TPH; Folin-Ciocalteu method) contents and polyphenol oxidase (PPO; spectrophotometric method using catechol as a substrate), peroxidase (POD; guaiacol substrate), and phenylalanine ammonia-lyase (PAL; phenylalanine substrate) activities were determined in photosynthetic and vascular tissue from outer and inner leaves. Unprocessed photosynthetic and inner leaf tissues had significantly higher (P < 0.05) levels of protein and TPH and PPO and POD activities than vascular and outer leaf tissues. PAL activities (on a fresh weight basis) were similar in all tissues. Changes in browning (light reflectance measurement) and phenolic metabolism in all four tissue types were observed during aerobic storage at 5 degrees C over 10 days. PAL activity increased in all tissues after 1-2 days of storage and then gradually decreased. POD activity also increased steadily for the storage duration. Protein content and PPO activity remained constant. Edge browning (measured with a Minolta Chroma Meter) and TPH increased in all tissues, especially in outer vascular tissue. Cut photosynthetic and vascular tissues washed at 4 and 47 degrees C with and without 100 microg mL(-1) chlorine for 3 min were analyzed during 7 days in storage at 5 degrees C. Enzyme activities and accumulation of phenolics in all tissues washed at 47 degrees C were significantly (P < 0.05) lower compared to controls or tissues washed at 4 degrees C. Chlorine had no additional effect at 47 degrees C but significantly (P < 0.05) reduced browning and accumulation of phenolics in lettuce washed at 4 degrees C. These results showed that inherent differences between tissues affect phenolic metabolism and browning in stored, fresh-cut lettuce.

Survival and growth of Listeria monocytogenes and Escherichia coli O157:H7 in ready-to-eat iceberg lettuce washed in warm chlorinated water.

Cut iceberg lettuce inoculated with Escherichia coli O157:H7 and Listeria monocytogenes before and after washing for 3 min in cold (4 degrees C) and warm (47 degrees C) water containing 100 mg/liter total chlorine was stored at I and 10 degrees C in oxygen-permeable film packages (6,000 to 8,000 cc/m2/24 h). Cold chlorinated water was detrimental to the survival of E. coli O157: H7 and L. monocytogenes at both storage temperatures. In contrast, washing in warm chlorinated water favored the growth of both pathogens in lettuce stored at 10 degrees C. There was no evidence of a relationship between the magnitude of spoilage microflora and the fate of either bacterium.