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Butyl acetate and yeasts interact in adhesion and germination of Botrytis cinerea conidia in vitro and in fungal decay of golden delicious apple.

Butyl acetate is a volatile aroma and flavor compound in apple. Conidia of three strains of Botrytis cinerea, a fungus that causes decay of apple fruit in postharvest storage, had greater adhesion to and greater germination on polycarbonate membrane filters on water inside sealed 500 cc glass jars that were injected with 4 microliters butyl acetate than conidia not so exposed. Conidial germination was highly correlated with conidial adhesion. The yeasts Sporobolomyces roseus and Cryptococcus laurentii, but not Saccharomyces cerevisiae, reduced the adhesion and germination promoting effect of butyl acetate. Conidia did not readily utilize butyl acetate as a food source, as shown by lack of tetrazolium violet reduction, whereas S. roseus and C. laurentii, but not S. cerevisiae did. Butyl acetate added to suspensions of conidia increased the electrical conductivity of the suspensions and increased the loss of 14C from 14C-labeled conidia compared to conidia unexposed to butyl acetate. Uptake of [14C]glucose by conidia was not increased by butyl acetate. Wounds of Golden Delicious apples inoculated with conidia (strain F-J-4) in a dilute solution of butyl acetate had greater decay than unexposed wounds. S. roseus and C. laurentii, but not S. cerevisiae, added with the conidia decreased the incidence or size of decay. Results indicated that butyl acetate increased conidial adhesion, stimulating conidial germination, and some yeasts can reduce this effect.

Acetates↗

AFLP-derived SCARs facilitate construction of a 1.1 Mb sequence-ready map of a region that spans the Vf locus in the apple genome.

The availability of high-density anchored markers is a prerequisite for reliable construction of a deep coverage BAC contig, which leads to creation of a sequence-ready map in the target chromosomal region. Unfortunately, such markers are not available for most plant species, including woody perennial plants. Here, we report on an efficient approach to build a megabase-size sequence-ready map in the apple genome for the Vf region containing apple scab resistance gene(s) by targeting AFLP-derived SCAR markers to this specific genomic region. A total of 11 AFLP-derived SCAR markers, previously tagged to the Vf locus, along with three other Vf-linked SCAR markers have been used to screen two apple genome BAC libraries. A single BAC contig which spans the Vf region at a physical distance of approximately 1,100 kb has been constructed by assembling the recovered BAC clones, followed by closure of inter-contig gaps. The contig is approximately 4 x deep, and provides a minimal tiling path of 16 contiguous and overlapping BAC clones, thus generating a sequence-ready map. Within the Vf region, duplication events have occurred frequently, and the Vf locus is restricted to the ca. 290 kb region covered by a minimum of three overlapping BAC clones.

Ascomycota↗

Sustainability of three apple production systems.

Escalating production costs, heavy reliance on non-renewable resources, reduced biodiversity, water contamination, chemical residues in food, soil degradation and health risks to farm workers handling pesticides all bring into question the sustainability of conventional farming systems. It has been claimed, however, that organic farming systems are less efficient, pose greater health risks and produce half the yields of conventional farming systems. Nevertheless, organic farming became one of the fastest growing segments of US and European agriculture during the 1990s. Integrated farming, using a combination of organic and conventional techniques, has been successfully adopted on a wide scale in Europe. Here we report the sustainability of organic, conventional and integrated apple production systems in Washington State from 1994 to 1999. All three systems gave similar apple yields. The organic and integrated systems had higher soil quality and potentially lower negative environmental impact than the conventional system. When compared with the conventional and integrated systems, the organic system produced sweeter and less tart apples, higher profitability and greater energy efficiency. Our data indicate that the organic system ranked first in environmental and economic sustainability, the integrated system second and the conventional system last.

Agriculture↗

Optical properties and contribution of cuticle to UV protection in plants: experiments with apple fruit.

To assess the UV-screening capacity of plant surface structures, the optical properties of isolated cuticle and detached peel of apple fruit (Malus domestica Borkh., cv. Antonovka) have been studied. It was found that the cuticle exhibits considerable scattering of UV radiation, negligible absorption between 500-800 nm and attenuates UV radiation: on average, cuticular transmittance of non-reflected light amounts to 35.7 +/- 20.2 and 14.2 +/- 7.1% at 375 and 300 nm, respectively. The principal UV-A absorbers in the cuticle were identified as quercetin glycosides with an in vivo absorption maximum near 375 nm and content ranging from 10 to 70 nmol cm(-2). On the shaded side of apple fruit, both UV-A and UV-B absorption by the peel is, to a large extent, governed by cuticular phenolics, whereas on the sunlit surface, the absorption of the peel in the UV-A range is determined mainly by vacuolar peel flavonoids. It is concluded that a massive build-up of flavonoids in the peel cells located just below the cuticle, resulting in trapping of radiation in a broad spectral range, plays a dominant role in the long-term adaptation of apple fruit to elevated levels of solar radiation.

Fruit↗

Cold storage affects survival and growth of Erwinia amylovora on the calyx of apple.

AIMS: To determine the effect of cold storage on the survival of Erwinia amylovora. METHODS AND RESULTS: The survival of E. amylovora was assessed during storage at 2 degrees C. Populations of E. amylovora inoculated into phosphate-buffered saline remained static, whereas in nutrient media populations increased at low temperatures. In contrast, populations of E. amylovora on tissue in the apple calyx decreased during cold storage. CONCLUSIONS: Erwinia amylovora has the ability, in nutrient media, to multiply at low temperatures. However, populations of E. amylovora on tissue in the apple calyx decrease with the time spent in cold storage. SIGNIFICANCE AND IMPACT OF THE STUDY: Cold storage of apples will provide assurance that mature fruit from orchards, free of fire blight, or even with low levels of fire blight, may be exported with a negligible risk of introducing the disease into countries free of fire blight.

Cold Temperature↗

Factor XI apple domains and protein dimerization.

The coagulation protease zymogen factor (F)XI is a disulfide bond-linked homodimer, a configuration that is necessary for protein secretion and function. The non-catalytic portion of the FXI polypeptide contains four repeats called apple domains (A1-A4). It is clear that FXI A4 plays a key role in dimer formation, however, the importance of other apple domains to this process has not been examined. We prepared recombinant FXI molecules in which apple domains were exchanged with those of the structurally homologous monomeric protein prekallikrein (PK). As expected, FXI/PK chimeras containing FXI A4 are dimers, while those with PK A4 are monomers. FXI A4 contains cysteine at position 321 that forms the interchain disulfide bond, while Cys321 in PK is unavailable for interchain bond formation because it is paired with Cys326. FXI/PK chimeras containing PK A4 were modified by changing Cys326 to glycine, leaving Cys321 unpaired (PKA4-Gly326). FXI with a PK A4 domain is a monomer, however, introducing PKA4-Gly326 results in a disulfide bond-linked dimer. This indicates that dimer formation can occur in the absence of FXI A4. In proteins containing PKA4-Gly326, replacing FXI A3 with PK A3 partially interferes with dimer formation, while substitution of A2, or A2 and A3 prevents dimer formation. PKA4-Gly326 cannot induce the native PK molecule to dimerize. The data indicate that FXI A2 and A3 make contributions to dimer formation. As these domains are involved in activities that require dimeric protein, it seems reasonable that they stabilize this conformation.

Binding Sites↗

Excellent long-term outcome for survivors of apple peel atresia.

BACKGROUND: Apple peel atresia is the rarest type of small bowel atresia. Because of its rare occurrence and high mortality rate, little is known about the long-term outcome of these children. METHODS: The patient charts, operative reports, and office notes of 15 children with apple peel atresia from 6 pediatric surgical centers in the Netherlands were reviewed. Long-term follow-up was assessed through review of office notes and through questionnaires. RESULTS: The median age at the time of operation was 1.5 days. Postoperatively, 53% suffered from cholestasis, and 40% were septic. Three patients died (20%). At follow-up at a median age of 24 months, 1 child showed growth retardation and 2 children suffered from short bowel syndrome. At the time of the questionnaire, all children showed normal growth and development. CONCLUSIONS: Even though children with apple peel atresia often suffer serious morbidity like short bowel syndrome and sepsis during the postoperative course, late morbidity turned out to be low. If the patients survive the operative and direct postoperative period, and survive the morbidity associated with malnutrition and the long-term use of total parenteral nutrition, they have a good chance of having normal bowel function with normal growth and development.

Female↗

Double-blind, randomized evaluation of clinical efficacy and tolerability of an apple pectin-chamomile extract in children with unspecific diarrhea.

BACKGROUND: Acute diarrhea is one of the most common childhood diseases. The main aim of therapy is oral rehydration, mostly using a glucose-electrolyte solution. Results from a previous study (DIALOG I) investigating adjuvant treatment with a medicinal preparation containing an apple pectin-chamomile extract (Diarrhoesan) indicated a significantly reduced duration of diarrhea in children. OBJECTIVES: The objective of the present clinical placebo-controlled, double-blind study (DIALOG II) was to assess the clinical efficacy and tolerability of the apple pectin-chamomile extract on a larger number of patients. METHODS: The investigation was designed as a multicenter, randomized, double-blind, placebo-controlled parallel study enrolling 255 patients presenting with acute diarrhea. Patients were aged between 6 months and 6 years and treated on an outpatient basis with either an apple pectin-chamomile preparation or placebo. As a basic medication, each child received a glucose-electrolyte solution on the first day of treatment. RESULTS: The primary outcome (primary efficacy parameter) included a combined analysis of stool frequency, stool The statistical analysis revealed a superior efficacy of the tested preparation over placebo with a significantly reduced stool frequency in the treatment group compared to the control group. The results were corroborated by efficacy assessment performed by investigators and patients. Treatment was well tolerated, with an incidence of adverse effects similar to placebo. CONCLUSION: These findings support the concept of a beneficial influence of the investigated vegetable extract in shortening the course of the disease and relieving associated symptoms.

Antidiarrheals↗

Identification of a factor IX binding site on the third apple domain of activated factor XI.

Activated factor XI (factor XIa) participates in blood coagulation by activating factor IX. Previous work has demonstrated that a binding site for factor IX is present on the noncatalytic heavy chain of factor XIa (Sinha, D., Seaman, F. S., and Walsh, P. N. (1987) Biochemistry 26, 3768-3775). Recombinant factor XI proteins were expressed in which each of the four apple domains of the heavy chain (designated A1 through A4) were individually replaced with the corresponding domain from the homologous but functionally distinct protease prekallikrein (PK). To identify the site of factor IX binding, the chimeric proteins were activated with factor XIIa and tested for their capacity to activate factor IX in plasma coagulation and purified protein assays. The chimera with the substitution in the third apple domain (factor XI/PKA3) had <1% of the coagulant activity of wild type factor XIa in a plasma coagulation assay, whereas the chimeras with substitutions in A1, A2, and A4 demonstrated significant activity (68-140% of wild type activity). The Km for activation of factor IX by factor XIa/PKA3 (12. 7 microM) is more than 30-fold higher than the Km for activation by wild type factor XIa or the other factor XI/PK chimeras (0.11-0.37 microM). Two monoclonal antibodies (2A12 and 11AE) that recognize epitopes on the factor XI A3 domain were potent inhibitors of factor IX activation by factor XIa, whereas antibodies against the A2 (1A6) and A4 (3G4) domains were poor inhibitors. The data indicate that a binding site for factor IX is present on the third apple domain of factor XIa.

Antibodies, Monoclonal↗

Mapping of the discontinuous H-kininogen binding site of plasma prekallikrein. Evidence for a critical role of apple domain-2.

Plasma prekallikrein, a zymogen of the contact phase system, circulates in plasma as heterodimeric complex with H-kininogen. The binding is mediated by the prekallikrein heavy chain consisting of four apple domains, A1 to A4, to which H-kininogen binds with high specificity and affinity (K(D) = 1.2 x 10(-8) M). Previous work had demonstrated that a discontinuous kininogen-binding site is formed by a proximal part located in A1, a distal part exposed by A4, and other yet unidentified portion(s) of the kallikrein heavy chain. To detect relevant binding segment(s) we recombinantly expressed single apple domains and found a rank order of binding affinity for kininogen of A2 > A4 approximately A1 > A3. Removal of single apple domains in prekallikrein deletion mutants reduced kininogen binding by 21 (A1), 64 (A2), and 24% (A4), respectively, whereas deletion of A3 was without effect. Transposition of homologous A2 domain from prekallikrein to factor XI conferred high-affinity kininogen binding from the former to the latter. The principal role of A2 for H-kininogen docking to the prekallikrein heavy chain was further substantiated by the finding that cleavage of a single peptide bond in A2 drastically diminished the H-kininogen binding affinity. Furthermore, the epitope of monoclonal antibody PKH6 which blocks kallikrein-kininogen complex formation with an IC(50) of 8 nM mapped to the center portion of domain A2. Our data indicate that domain A2 and two flanking sequence segments of A1 and A4 form a discontinuous binding platform for H-kininogen on the prekallikrein heavy chain. Domain-specific antibodies directed to these critical sites efficiently interfered with contact phase-induced bradykinin release from H-kininogen.

Binding Sites↗

The toxoplasma micronemal protein MIC4 is an adhesin composed of six conserved apple domains.

The initial stage of invasion by apicomplexan parasites involves the exocytosis of the micronemes-containing molecules that contribute to host cell attachment and penetration. MIC4 was previously described as a protein secreted by Toxoplasma gondii tachyzoites upon stimulation of micronemes exocytosis. We have microsequenced the mature protein, purified after discharge from micronemes and cloned the corresponding gene. The deduced amino acid sequence of MIC4 predicts a 61-kDa protein that contains 6 conserved apple domains. Apple domains are composed of six spacely conserved cysteine residues which form disulfide bridges and are also present in micronemal proteins from two closely related apicomplexan parasites, Sarcocystis muris and Eimeria species, and several mammalian serum proteins, including kallikrein. Here we show that MIC4 localizes in the micronemes of all the invasive forms of T. gondii, tachyzoites, bradyzoites, sporozoites, and merozoites. The protein is proteolytically processed both at the N and the C terminus only upon release from the organelle. MIC4 binds efficiently to host cells, and the adhesive motif maps in the most C-terminal apple domain.

Amino Acid Sequence↗

Influence of grapefruit-, orange- and apple-juice consumption on urinary variables and risk of crystallization.

Alkalizing beverages are highly effective in preventing the recurrence of calcium oxalate (Ox), uric acid and cystine lithiasis. The aim of the present study was to evaluate the influence of grapefruit-juice and apple-juice consumption on the excretion of urinary variables and the risk of crystallization in comparison with orange juice. All investigations were carried out on nine healthy female subjects without any history of stone formation and aged 26-35 years. Each juice was tested in a 5 d study. During the study, the subjects received a standardized diet. Fluid intake of 2.75 litres was composed of 2.25 litres neutral mineral water, 0.4 litre coffee and 0.1 litre milk. On the fourth and fifth day 0.5 litre mineral water was partly substituted by 0.5 or 1.0 litre juice under investigation respectively. The influence on urinary variables was evaluated in 24 h urine samples. In addition, the BONN risk index of CaOx, relative supersaturation (RS)CaOx crystallization was determined. Due to an increased pH value and an increased citric acid excretion after consumption of each juice, the RSCaOx decreased statistically significantly (P<0.05) for grapefruit juice, but not significantly for orange and apple juice. The BONN risk index yielded a distinct decrease in the crystallization risk. We showed that both grapefruit juice and apple juice reduce the risk of CaOx stone formation at a magnitude comparable with the effects obtained from orange juice.

Adult↗

Co-occurrence of patulin and citrinin in Portuguese apples with rotten spots.

Patulin and citrinin are mycotoxins produced by certain fungi mainly belonging to Penicillium and Aspergillus and may be detected in mouldy fruits and fruit products. The data presented here refer to the simultaneous occurrence of patulin and citrinin in 351 samples of seven different varieties of apples with small rotten areas (Casanova, Golden Delicious, Red Delicious, Reineta, Richared, Rome Beauty, Starking). A rapid multidetection thin layer chromatography (TLC) method was used. The minimum detectable concentrations of patulin and citrinin were 120-130 and 15-20 microg kg(-1) respectively. The percentage contamination with patulin only was higher (68.6%) than that with citrinin only (3.9%). Patulin and citrinin (19.6%) were also detected simultaneously. The highest mean patulin content was 80.50 mg kg(-1) for the Richared variety, but the mean level of citrinin was lower. The lowest mean contaminations of patulin were found in Rome Beauty, Red Delicious and Reineta, ranging from 3.06 to 5.37 mg kg(-1). All analysed apples varieties had low citrinin contamination, ranging from 0.32 to 0.92 mg kg(-1). These findings indicate that there may be a risk of human exposure to patulin through the consumption of juices and jams manufactured with apples with small rotten areas.

Chromatography, Thin Layer↗

Effect of surface waxes on the persistence of chlorpyrifos-methyl in apples, strawberries and grapefruits.

The effects of the cuticle and epicuticular waxes of grapefruit, strawberry and apple on the photodegradation and penetration of chlorpyrifos-methyl were studied. Photodegradation experiments were conducted by exposing the insecticide to the light of a xenon lamp in the presence of a film of wax extracted from the fruit surface. The half-life of chlorpyrifos-methyl irradiated in absence of waxes was 9.6 min. The half-lives of pesticide irradiated in the presence of wax extracts of apple, grapefruit and strawberry were 83, 34 and 26 min, respectively. In penetration studies, fruit with and without wax layers were treated with an aqueous suspension of pesticide. The penetration of the pesticide from the cuticle to the pulp was measured after 24 h. Samples without wax contained a higher total amount of insecticide than those with wax. No pesticide was detected in samples of apple and grapefruit pulp. Residues were detected in all fractions of strawberry. The waxes and cuticle appear to have some effect on the photodegradation and penetration of chlorpyrifos-methyl in fruit samples.

Chlorpyrifos↗

A limited survey of retail apple and grape juices for the mycotoxin patulin.

A retail survey (38 samples) of apple, grape and other juice-based products for the mycotoxin patulin has been carried out using HPLC analysis. Positive results were confirmed by GC-MS of the TMS derivative using chemical ionization selected ion monitoring. No grape juice or mixed juice product contained detectable patulin (limit 5 micrograms/l), nor did the majority (58%) of the apple juices. Low levels of patulin (in the range 5-10 micrograms/l) were present in six apple juices and the remaining four positive samples contained 16, 18, 30 and 56 micrograms/l.

Beverages↗

The influence of deck storage and initial processing on patulin levels in apple juice.

Patulin, a secondary metabolite produced by Penicillium expansum and some other fungal species, is a common contaminant of ripened apples used for the production of apple juice concentrates. The limited availability of suitable storage facilities may result in fruit being subjected to storage in the open ('deck storage') for extended periods of time, prior to processing. A study was conducted to determine the influence that deck storage and subsequent initial processing practices had on patulin levels in freshly pressed juice. Over the study period, triplicate samples were collected at four strategic processing points from individual consignments of Granny Smith apples deck-stored for 7, 15 and 33 days, respectively. Over the study period, mean patulin levels in non-processed fruit increased from 90 to 2445 ng/g, respectively, but decreased to between 75 and 695 ng/g, respectively, following a water wash step. Subsequent removal of rotten/damaged fruit decreased patulin levels further (to between 55 and 405 ng/g, respectively), although the numerical decreases between sampling points were not shown to be statistically significant (P > 0.05). However, patulin levels were significantly higher (P < 0.05) in the rejected rotten/damaged fruit (mean levels ranged from 1120 to 6235 ng/g, respectively). P. expansum was the major patulin-producing fungus isolated from the juice samples. The mycological analyses tended to support the chemical data, in that removal of the rotten/damaged fractions significantly reduced total fungal counts in the juice samples.

Beverages↗

Physicochemical, nutritional and sensory quality of stirred 'dwarf' golden apple (Spondias cytherea Sonn) yoghurts.

The dwarf golden apple (Spondias cytherea Sonn) is an exotic fruit which is mainly consumed in its fresh form but could be processed. The objectives of the study were to investigate the effects of adding golden apple nectar on the physicochemical and sensory quality of stirred yoghurts from cow's milk. Yoghurts with 15% and 20% golden apple nectar were more (P>0.01) liked than the control (0% nectar) yoghurt in all sensory attributes. The appearance and body attribute differed (P<0.01) between yoghurts with 15% and 20% nectar. The overall sensory quality of these yoghurts was rated very good to excellent. There were significant (P>0.05) changes, in pH, lactic acid, consistency, colour, lactic acid bacteria and yeasts and moulds on storage at 4 degrees C for 4 weeks. By week 4 of storage, yoghurts developed a buttery smell and were less dark and yellow. A 226 g yoghurt serving provided an excellent source of phosphorus and was good in protein.

Adult↗

Review of apple flavor--state of the art.

The retention flavor is of utmost importance during the harvesting, handling and/or further processing of apples. The complexity of the natural flavor of the apple or essence derived therefrom is attributed in part to the variety, post-harvest treatment, storage, and process manipulation. Some 266 volatile components isolated from apples include alcohols, esters, aldehydes, ketones, ethers, acids, bases, acetals, and hydrocarbons. This review is a survey of the literature published during the last 10 to 15 years and includes the contribution of peel to flavor, postharvest quality indices related to flavor, storage and dehydration effects on flavor, juice extraction methods, sensory evaluation, and volatile separation and identification techniques.

Acetals↗