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At least 451 records · Page 25Linked to original sources

Identification and molecular properties of a 306 nucleotide viroid associated with apple dimple fruit disease.

A new viroid associated with an apple fruit disorder similar to, but more severe than, the dapple apple disease induced in some varieties by apple scar skin viroid (ASSVd) has been found. The new viroid, tentatively termed apple dimple fruit viroid (ADFVd), is a circular RNA of 306 nucleotides which adopts a quasi-rod-like conformation of minimum free energy. It contains the core nucleotides of the central conserved region (CCR) of the ASSVd group as well as the terminal conserved region (TCR) present in all members of the ASSVd and potato spindle tuber viroid (PSTVd) monophyletic groups. ADFVd has the highest sequence similarity with ASSVd and the 294 nucleotide citrus viroid CVd-IIIb sharing with the latter an almost identical left terminal domain. The right- and left-hand termini of ADFVd are formed by almost perfect duplications of sequences found in the CCR upper and lower strands, respectively, of PSTVd and closely related viroids.

Base Sequence↗

Overexpression of polygalacturonase in transgenic apple trees leads to a range of novel phenotypes involving changes in cell adhesion.

Polygalacturonases (PGs) cleave runs of unesterified GalUA that form homogalacturonan regions along the backbone of pectin. Homogalacturonan-rich pectin is commonly found in the middle lamella region of the wall where two adjacent cells abut and its integrity is important for cell adhesion. Transgenic apple (Malus domestica Borkh. cv Royal Gala) trees were produced that contained additional copies of a fruit-specific apple PG gene under a constitutive promoter. In contrast to previous studies in transgenic tobacco (Nicotiana tabacum) where PG overexpression had no effect on the plant (K.W. Osteryoung, K. Toenjes, B. Hall, V. Winkler, A.B. Bennett [1990] Plant Cell 2: 1239-1248), PG overexpression in transgenic apple led to a range of novel phenotypes. These phenotypes included silvery colored leaves and premature leaf shedding due to reduced cell adhesion in leaf abscission zones. Mature leaves had malformed and malfunctioning stomata that perturbed water relations and contributed to a brittle leaf phenotype. Chemical and ultrastructural analyses were used to relate the phenotypic changes to pectin changes in the leaf cell walls. The modification of apple trees by a single PG gene has offered a new and unexpected perspective on the role of pectin and cell wall adhesion in leaf morphology and stomatal development.

Cell Adhesion↗

An allele of the ripening-specific 1-aminocyclopropane-1-carboxylic acid synthase gene (ACS1) in apple fruit with a long storage life.

An allele of the 1-aminocyclopropane-1-carboxylic acid (ACC) synthase gene (Md-ACS1), the transcript and translated product of which have been identified in ripening apples (Malus domestica), was isolated from a genomic library of the apple cultivar, Golden Delicious. The predicted coding region of this allele (ACS1-2) showed that seven nucleotide substitutions in the corresponding region of ACS1-1 resulted in just one amino acid transition. A 162-bp sequence characterized as a short interspersed repetitive element retrotransposon was inserted in the 5'-flanking region of ACS1-2 corresponding to position -781 in ACS1-1. The XhoI site located near the 3' end of the predicted coding region of ACS1-2 was absent from the reverse transcriptase-polymerase chain reaction product, revealing that exclusive transcription from ACS1-1 occurs during ripening of cv Golden Delicious fruit. DNA gel-blot and polymerase chain reaction analyses of genomic DNAs showed clearly that apple cultivars were either heterozygous for ACS1-1 and ACS1-2 or homozygous for each type. RNA gel-blot analysis of the ACS1-2 homozygous Fuji apple, which produces little ethylene and has a long storage life, demonstrated that the level of transcription from ACS1-2 during the ripening stage was very low.

Alleles↗

A study of ethylene in apple, red raspberry, and cherry.

High ethylene levels were associated with flower abscission in apple (Malus sylvestris) and cherry (Prunus avium and Prunus cerasus), "June drop" of immature cherries, and harvest drop of apple and red raspberry (Rubus idaeus). However, an increase in ethylene content was not associated with June drop of apples and harvest drop of cherries. During the period of fruit ripening on the plant, the largest increases in ethylene occurred in apple flesh and red raspberry receptacular tissue. Ethylene remained low throughout the period of sweet and tart cherry ripening. The data obtained indicated marked ethylene gradients between adjacent tissues. Increases of ethylene in some tissues may have resulted from ethylene diffusion from adjacent tissues containing high levels of ethylene.

Journal Article↗

Ethylene-forming Systems in Etiolated Pea Seedling and Apple Tissue.

Auxin-induced ethylene formation in etiolated pea (Pisum sativum L. var. Alaska) stem segments was inhibited by inhibitors of RNA and protein synthesis. Kinetics of the inhibitions is described for actinomycin D, cordycepin, alpha-amanitin, and cycloheximide. alpha-Amanitin was the most potent and fast-acting inhibitor, when added before induction or 6 hours after induction of the ethylene-forming system. The ethylene-forming system of postclimacteric apple (Malus sylvestris L.) tissue, which is already massively induced, was not further stimulated by auxin. Ethylene production in apples was inhibited least by alpha-amanitin and most by actinomycin D. The relative responses of the ethylene system in apples to RNA inhibitors were different from the ethylene system of pea stems. However, the protein synthesis inhibitor, cycloheximide, appeared to act equally in both tissue systems. The effect of cycloheximide on ethylene production in postclimacteric apple tissue, already producing large quantities of ethylene, suggests a dynamic regulating system for the synthesis and degradation of the ethylene-forming system.

Journal Article↗

Ultrastructural changes in the cell walls of ripening apple and pear fruit.

Ultrastructural changes in the cell walls of "Calville de San Sauveur" apples (Malus sylvestris Mill) and "Spadona" pear (Pyrus communis L.) fruit were followed during ripening. In apple, structural alterations in cell walls became apparent at advanced stages of softening and showed predominantly dissolution of the middle lamella. In pears softening was also associated with the dissolution of the middle lamella, and in addition a gradual disintegration of fibrillar material throughout the cell wall. In fully ripe fruit almost all of the fibrillar arrangement in the cell wall was lost. Application of enzyme solutions containing polygalacturonase and cellulase to tissue discs from firm pear fruit led to ultrastructural changes observed in naturally ripening pears. In apple polygalacturonase alone was sufficient to dissolve the middle lamella region of the cell walls, as was also found to occur in naturally ripening fruit. In both apple and pear the cell wall areas containing plasmodesmata maintained their structural integrity throughout the ripening process. At advanced stages of ripening vesicles appeared in the vicinity of plasmodesmata.

Journal Article↗

Purification and characterization of 1-aminocyclopropane-1-carboxylate synthase from apple fruits.

1-Aminocyclopropane-1-carboxylate (ACC) synthase, a key enzyme in ethylene biosynthesis, was isolated and partially purified from apple (Malus sylvestris Mill.) fruits. Unlike ACC synthase isolated from other sources, apple ACC synthase is associated with the pellet fraction and can be solubilized in active form with Triton X-100. Following five purification steps, the solubilized enzyme was purified over 5000-fold to a specific activity of 100 micromoles per milligram protein per hour, and its purity was estimated to be 20 to 30%. Using this preparation, specific monoclonal antibodies were raised. Monoclonal antibodies against ACC synthase immunoglobulin were coupled to protein-A agarose to make an immunoaffinity column, which effectively purified the enzyme from a relatively crude enzyme preparation (100 units per milligram protein). As with the tomato enzyme, apple ACC synthase was inactivated and radiolabeled by its substrate S-adenosyl-l-methionine. Apple ACC synthase was identified to be a 48-kilodalton protein based on the observation that it was specifically bound to immunoaffinity column and it was specifically radiolabeled by its substrate S-adenosyl-l-methionine.

Journal Article↗

Role of lipopeptides produced by Bacillus subtilis GA1 in the reduction of grey mould disease caused by Botrytis cinerea on apple.

AIM: Test of Bacillus subtilis strain GA1 for its potential to control grey mould disease of apple caused by Botrytis cinerea. METHODS AND RESULTS: GA1 was first tested for its ability to antagonize in vitro the growth of a wide variety of plant pathogenic fungi responsible for diseases of economical importance. The potential of strain GA1 to reduce post-harvest infection caused by B. cinerea was tested on apples by treating artificially wounded fruits with endospore suspensions. Strain GA1 was very effective at reducing disease incidence during the first 5 days following pathogen inoculation and a 80% protection level was maintained over the next 10 days. Treatment of fruits with an extract of GA1 culture supernatant also exerted a strong preventive effect on the development of grey mould. Further analysis of this extract revealed that strain GA1 produces a wide variety of antifungal lipopeptide isomers from the iturin, fengycin and surfactin families. A strong evidence for the involvement of such compounds in disease reduction arose from the recovery of fengycins from protected fruit sites colonized by bacterial cells. CONCLUSIONS: The results presented here demonstrate that, despite unfavourable pH, B. subtilis endospores inoculated on apple pulp can readily germinate allowing significant cell populations to establish and efficient in vivo synthesis of lipopeptides which could be related to grey mould reduction. SIGNIFICANCE AND IMPACT OF THE STUDY: This work enables for the first time to correlate the strong protective effect of a particular B. subtilis strain against grey mould with in situ production of fengycins in infected sites of apple fruits.

Antifungal Agents↗

Hypersensitivity to apple and carrot can be reliably detected with fresh material.

Hypersensitivity to birch pollen was linked with allergy to apple and carrot, apple seeds and carrot seeds and also with their fractions. Skin reactions among other atopics were rare, and non-atopic patients showed no reactions to fresh fruits and vegetables. Apple and carrot as well as apple seeds and carrot seeds were fractionated using column chromatography. Two major fractions were obtained, one with sugar and proteins, and one with proteins, the latter giving skin reactions more often than the former. The correlation between clinical symptoms and skin test results with the fractionated material was somewhat lower than that obtained with the non-fractionated fresh material. The allergens of fruits and vegetables might be proteins (enzymes).

Adolescent↗

Immunological investigation of possible structural similarities between pollen antigens and antigens in apple, carrot and celery tuber.

Crossed line immunoelectrophoresis (CLIE) was used to show that a minor allergen present in birch pollen and another present in timothy pollen shared common epitopes with antigens in apple, carrot and celery tuber. Major pollen allergens were not involved. Structural similarities were also noticed in some mugwort pollen antigens and antigens in apple, carrot and celery, but none of these mugwort antigens acted as an allergen. In crossed immunoelectrophoresis (CIE) of samples of apple, carrot or celery extracts towards antibodies against birch pollen, grass pollen or mugwort pollen, four to 14 distinct precipitates were observed and shown to be specific. In crossed radioimmunoelectrophoresis (CRIE) none of the precipitates could be shown to act as an allergen. Using allergen detection on nitrocellulose paper, the apple, carrot and celery extracts were shown to possess IgE-binding capacity.

Allergens↗

An investigation of the possible immunological relationship between allergen extracts from birch pollen, hazelnut, potato and apple.

In a retrospective study on selected group of patients, the coincidence of birch pollen allergy and a clinically relevant positive prick test reaction to apples and potatoes was confirmed. Immunochemical comparison using the crossed line immunoelectrophoresis technique (CLIE) confirmed partial identity between birch pollen and hazelnut. By the same method no partial immunological identity between birch pollen and extracts and fresh peel from apples and potatoes was found. However, both apples and potatoes gave rise to non-immunological affinity precipitates. On this basis it is discussed, whether the active agents from apples and potatoes were extremely labile allergens, plant lectins, or low molecular weight substances with a direct or indirect histamine release effect.

Dermatitis, Contact↗

Biocontrol of the food-borne pathogens Listeria monocytogenes and Salmonella enterica serovar Poona on fresh-cut apples with naturally occurring bacterial and yeast antagonists.

Fresh-cut apples contaminated with either Listeria monocytogenes or Salmonella enterica serovar Poona, using strains implicated in outbreaks, were treated with one of 17 antagonists originally selected for their ability to inhibit fungal postharvest decay on fruit. While most of the antagonists increased the growth of the food-borne pathogens, four of them, including Gluconobacter asaii (T1-D1), a Candida sp. (T4-E4), Discosphaerina fagi (ST1-C9), and Metschnikowia pulcherrima (T1-E2), proved effective in preventing the growth or survival of food-borne human pathogens on fresh-cut apple tissue. The contaminated apple tissue plugs were stored for up to 7 days at two different temperatures. The four antagonists survived or grew on the apple tissue at 10 or 25 degrees C. These four antagonists reduced the Listeria monocytogenes populations and except for the Candida sp. (T4-E4), also reduced the S. enterica serovar Poona populations. The reduction was higher at 25 degrees C than at 10 degrees C, and the growth of the antagonists, as well as pathogens, increased at the higher temperature.

Ascomycota↗

Effect of an apple brandy from Normandy and of ethanol on epithelial cell proliferation in the esophagus of rats.

The effects on cell population kinetics in the distal esophagus of an apple brandy from Normandy (Calvados), of ethanol diluted at a similar concentration, and of saline were investigated in rats. Animals were killed at 0, 6, 12, 18 and 24 h after a single intraesophageal instillation, and at 3 weeks after chronic administration of these solutions. In vivo labeling with [3H]-thymidine and autoradiography were carried out to determine the proliferative parameters. Vincristine sulfate was used to increase metaphase figures during the chronic experiment. Instillation of saline did not modify the parameters under study. Single instillations of both apple brandy and 56 degrees ethanol were followed by a significant increase in labeling (p less than 0.01) and mitotic indices (p less than 0.01). Chronic administration of both apple brandy and 56 degrees ethanol increased the labeling index (p less than 0.01) as well as the metaphase index (p less than 0.01). The proliferative responses after the apple brandy and after 56 degrees ethanol were similar in amplitude. These data suggest that ethanol alone may explain the mitogenic effect of Calvados in the esophagus.

Alcoholic Beverages↗

Characterization of the 18-kDa apple allergen by two-dimensional immunoblotting and microsequencing.

A low-temperature extract taken from Golden Delicious apples was separated by two-dimensional polyacrylamide gel electrophoresis. By means of two-dimensional immunoblotting with patients' serum containing IgE specific to Bet v I, a rabbit polyclonal antiserum raised against Bet v I, and two Bet v I specific monoclonal antibodies, epitopes cross-reactive to Bet v I were identified on an apple allergen with a molecular mass of 18 kDa and pI 5.5. Furthermore, certain antibody reactivities with 4 isoproteins of a molecular mass of 16 kDa and pIs ranging from 4.9 to 5.5 were observed, which may indicate the presence of Bet v I related epitopes on these proteins. Based on 26 amino acid residues, N-terminal sequencing of the 18-kDa apple allergen revealed 62% sequence identity between Bet v I from birch pollen and the apple allergen. Our results therefore support the view that both proteins express common as well as non-related IgE-reactive epitopes.

Allergens↗

Carvacrol and p-cymene inactivate Escherichia coli O157:H7 in apple juice.

BACKGROUND: Outbreaks of food poisoning associated with drinking un-pasteurised apple juice contaminated with enterohaemorrhagic Escherichia coli O157:H7 are a cause of serious illness and occasionally death. Whilst a well-established heat process (pasteurisation) will readily eliminate the pathogen, some consumers are demanding more fresh-like foods that have not been subjected to processing methods that are perceived as severe and may lead to loss of flavour and vitamins. Therefore, alternative methods are being investigated to replace pasteurisation and improve the safety of minimally-processed juices. The addition of natural antimicrobial substances such as the phenolic substances carvacrol and p-cymene (derived from the essential oils of herbs and spices) provides a potential new route to assure safety and extend the shelf-life of raw fruit juices. The aim of this study was to evaluate the addition of very low concentrations (0.25-1.25 mM) of carvacrol and p-cymene both individually and in combination as a novel means of controlling Escherichia coli O157:H7 in un-pasteurised apple juice. RESULTS: When inoculated at a level of 4 log CFU/ml into un-pasteurised apple juice (pH 3.20 +/- 0.06), Escherichia coli O157:H7 survived for up to 3 and 19 days at 25 degrees and 4 degrees C, respectively. Treatment of the juice with 1.25 mM carvacrol or p-cymene reduced the numbers of E. coli O157:H7 to undetectable levels within 1-2 days at both storage temperatures. The effective concentrations of carvacrol could be reduced even further by combining it at 0.5 mM with cymene at 0.25 mM. The phenolic compounds were biocidal against both spoilage yeasts and E. coli O157:H7 thereby increasing the shelf-life and improving the safety of un-pasteurised apple juice, particularly when stored at chill temperatures. CONCLUSION: The results showed that the natural antimicrobial compounds carvacrol and p-cymene could potentially be used to extend the shelf life and improve the safety margins in un-pasteurised chilled fruit juices.

Anti-Bacterial Agents↗

Subcellular location of polyphenol oxidase in apples.

The location of polyphenol oxidase (PPO) in cells of apple fruit was examined by immunohistochemistry and subcellular fractionation. In mature apple fruits, where vacuoles occupy most of the cells, PPO was detected immunochemically near the cell walls with use of anti-apple PPO antibodies. In cells of immature fruits and tissue culture, PPO was detected in organelles other than the vacuoles, probably in plastids. The plastid fraction was purified by density gradient ultracentrifugation, and the activities of PPO and marker enzymes of plastids were the highest in the plastids. Most apple PPO was in plastids, as are other plant PPOs, and some of the protein was solubilized and proteolyzed during ripening and storage.

Blotting, Western↗

Inhibitory effects of apple polyphenol on induced histamine release from RBL-2H3 cells and rat mast cells.

The anti-allergic activities of polyphenol fractions extracted from immature fruits of apple (Rosaceae, Malus sp.) were evaluated by in vitro assays. A crude apple polyphenol (CAP) fraction, which had been obtained from the juice of immature apples by reverse-phase column chromatography, was further purified by LH-20 column chromatography to obtain an apple condensed tannin (ACT) fraction consisting of linear oligomeric epicatechins from the dimer to pentadecamer. ACT strongly inhibited the release of histamine from rat basophilic leukemia (RBL-2H3) cells stimulated by the antigen-stimulation and from rat peritoneal mast cells stimulated by compound 48/80. The IC50 values for histamine release were 30 micrograms/ml and 25 micrograms/ml, respectively. ACT also inhibited hyaluronidase activity and the increase in intracellular free calcium concentration in RBL-2H3 cells stimulated with the antigen. These results suggest that ACT affected early signal transduction including the calcium influx.

Animals↗

Automated detection of fecal contamination of apples by multispectral laser-induced fluorescence imaging.

Animal feces are a suspected source of contamination of apples by disease-causing organisms such as Echerichia coli O157. Laser-induced fluorescence was used to detect different amounts of feces from dairy cows, deer, and a dairy pasture applied to Red Delicious apples. One day after application, detection for 1:2 and 1:20 dilutions was nearly 100%, and for 1:200 dilutions (<15 ng of dry matter) detection was >80%. Detection after apples had been washed and brushed was lowest for pasture feces; detection for 1:2, 1:20, and 1:200 dilutions of feces was 100%, 30%, and 0%, respectively. This technology may encourage development of commercial systems for detecting fecal contamination of apples.

Animals↗