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Salinity Induced Limitations on Photosynthesis in Prunus salicina, a Deciduous Tree Species.

The response of photosynthetic CO(2) assimilation to salinization in 19 year old Prunus salicina was evaluated under field conditions for a 3 year period. The observed decline in CO(2) assimilation capacity was apparently related to increasing leaf chloride (Cl(-)) content, and independent of changes in leaf carbohydrate status. The response of net CO(2) assimilation (A) to leaf intercellular CO(2) partial pressure (C(i)) indicated that the reduction in the capacity for A with Cl(-) was not the result of decreased stomatal conductance but a consequence of nonstomatal inhibition. The nonstomatal limitations to CO(2) assimilation capacity, as determined by the response of A to C(i) and biochemical assay, were related to a decline in the activity of ribulose 1,5-bisphosphate carboxylase (Rubpcase) and the pool size of triose phosphate, ribulose 1,5-bisphosphate (Rubp) and phosphoglycerate with increasing salinity. Lack of agreement between the initial slope of the A to C(i) response curve and Rubpcase activity suggests the occurrence of heterogeneous stomatal apertures with the high salinity treatment (28 millimolar). Prolonged exposure to chloride salts appeared to increase the Rubp or Pi regeneration limitation, decrease Rubpcase activity and reduce leaf chlorophyll content. Observed changes in the biochemical components of CO(2) fixation may, in turn, affect total leaf carbohydrates, which also declined with time and salinity. The reduction in Rubpcase activity was apparently a consequence of a reduced Rubpcase protein level rather than either a regulatory or inhibitory effect.

Journal Article↗

Pattern of the Cyanide-Potential in Developing Fruits : Implications for Plants Accumulating Cyanogenic Monoglucosides (Phaseolus lunatus) or Cyanogenic Diglucosides in Their Seeds (Linum usitatissimum, Prunus amygdalus).

The absolute cyanide content of developing fruits was determined in Costa Rican wild lima beans (Phaseolus lunatus), oil flax (Linum usitatissimum), and bitter almonds (Prunus amygdalus). The cyanide potential (HCN-p) of the lima bean and the almond fruit began to increase shortly after anthesis and then stopped before fruit maturity. In contrast, the flax inflorescence had a higher HCN-p in absolute terms than the mature flax fruit. At all times of its development the bean fruit contained the monoglucosides linamarin and lotaustralin. The almond and the flax fruits contained, at anthesis, the monoglucosides prunasin, and linamarin and lotaustralin, respectively, while, at maturity, only the corresponding diglucosides amygdalin, and linustatin and neolinustatin, respectively, were present.

Journal Article↗

Immunocytochemical Localization of Mandelonitrile Lyase in Mature Black Cherry (Prunus serotina Ehrh.) Seeds.

Mandelonitrile lyase (MDL, EC 4.1.2.10), which catalyzes the reversible dissociation of (R)-(+)-mandelonitrile to benzaldehyde and hydrogen cyanide, was purified to apparent homogeneity from mature black cherry (Prunus serotina Ehrh.) seeds by conventional protein purification techniques. This flavoprotein is monomeric with a subunit molecular mass of 57 kilodaltons. Glycoprotein character was shown by its binding to the affinity matrix concanavalin A-Sepharose 4B with subsequent elution by alpha-methyl-d-glucoside. Upon chemical deglycosylation by trifluoromethanesulfonic acid, the molecular mass was reduced to 50.9 kilodaltons. Two-dimensional gel analysis of deglycosylated MDL revealed the presence of several subunit isoforms of similar molecular mass but differing slightly in isoelectric point. Polyclonal antibodies were raised in New Zealand white rabbits against deglycosylated and untreated MDL. Antibody titers were determined by enzyme linked immunosorbent and dot immunobinding assays, while their specificities were assessed by Western immunoblot analysis. Antibodies raised against untreated lyase recognized several proteins in addition to MDL. In contrast, antisera raised against deglycosylated MDL were monospecific and were utilized for developmental and immunocytochemical localization studies. SDS-PAGE and immunoblotting analysis of seed proteins during fruit maturation showed that MDL first appeared in seeds shortly after cotyledons began development. In cotyledon cells of mature seeds, MDL was localized primarily in the cell wall with lesser amounts in the protein bodies, whereas in endosperm cells, this labeling pattern was reversed. N-terminal sequence data was gathered for future molecular approaches to the question of MDL microheterogeneity.

Journal Article↗

Level of Abscisic Acid in Integuments, Nucellus, Endosperm, and Embryo of Peach Seeds (Prunus persica L. cv Springcrest) during Development.

Free abscisic acid (ABA) in integuments, nucellus, endosperm, and embryo was determined throughout seed development of peach (Prunus persica L. cv Springcrest). Quantification of ABA was performed using combined high performance liquid chromatography-radioimmunoassay based on a monoclonal antibody raised against free (S)-ABA. In the integuments and endosperm, ABA concentration remained constant during the first 100 days after anthesis and rose in the following days when fresh weight was rapidly decreasing. In the nucellus, the ABA concentration variation pattern paralleled that of tissue growth. ABA concentration in the embryo increased constantly with the growth of the tissues to reach a maximum at the last growth stage. The role of ABA in peach seeds is discussed in relation to the development of the different seed tissues.

Journal Article↗

Development of the Potential for Cyanogenesis in Maturing Black Cherry (Prunus serotina Ehrh.) Fruits.

Biochemical changes related to cyanogenesis (hydrogen cyanide production) were monitored during maturation of black cherry (Prunus serotina Ehrh.) fruits. At weekly intervals from flowering until maturity, fruits (or selected parts thereof) were analyzed for (a) fresh and dry weights, (b) prunasin and amygdalin levels, and (c) levels of the catabolic enzymes amygdalin hydrolase, prunasin hydrolase, and mandelonitrile lyase. During phase I (0-28 days after flowering [DAF]), immature fruits accumulated prunasin (mean: 3 micromoles/fruit) but were acyanogenic because they lacked the above enzymes. Concomitant with cotyledon development during mid-phase II, the seeds began accumulating both amygdalin (mean: 3 micromoles/seed) and the catabolic enzymes and were highly cyanogenic upon tissue disruption. Meanwhile, prunasin levels rapidly declined and were negligible by maturity. During phases II (29-65 DAF) and III (66-81 DAF), the pericarp also accumulated amygdalin, whereas its prunasin content declined toward maturity. Lacking the catabolic enzymes, the pericarp remained acyanogenic throughout all developmental stages.

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Cold Acclimation in Genetically Related (Sibling) Deciduous and Evergreen Peach (Prunus persica [L.] Batsch): I. Seasonal Changes in Cold Hardiness and Polypeptides of Bark and Xylem Tissues.

Seasonal patterns of proteins and of cold hardiness were characterized in bark and xylem tissues of genetically related (sibling) deciduous and evergreen peach (Prunus persica [L.] Batsch). In contrast with deciduous trees, which entered endodormancy and abscised leaves in the fall, evergreen trees retained their leaves and exhibited shoot elongation under favorable environmental conditions. A successive increase in the cold hardiness of bark and xylem was observed during the fall in both genotypes. This was followed by a subsequent decrease from midwinter to spring. Xylem tissue in both genotypes exhibited deep supercooling and a significant correlation (r = 0.99) between the midpoint of the low-temperature exotherm and the subzero temperature at which 50% injury occurred (assessed by electrolyte leakage) was noted. The maximum hardiness level attained in deciduous trees was more than twofold that of evergreens. Seasonal pattern of proteins from bark and xylem of the sibling genotypes was characterized by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Among other qualitative and quantitative changes, accumulation of a 19-kilodalton polypeptide in the bark of both genotypes was observed during fall followed by a decrease in spring. This polypeptide accumulated to higher levels in the deciduous peach compared with the evergreen. Additionally, a 16-kilodalton protein exhibited the same pattern in deciduous trees but not in the evergreen trees. Both the 19- and a 16-kilodalton bark proteins conform to the criteria of a bark storage protein. The relationship of seasonal changes in protein to cold hardiness and dormancy in these genetically related peach genotypes is discussed.

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Influence of phosphate compounds on certain fungi and their preservative effects on fresh cherry fruit (Prunus cerasus, L.).

Studies were conducted to ascertain the retarding effects of four phosphate compounds (sodium hexametaphosphate, sodium tripolyphosphate, sodium tetraphosphate, and tetrasodium pyrophosphate) on molding of fresh cherries (Prunus cerasus, L.). In vitro studies on their antimycotic effects against the most common fungal spoilers, Penicillium expansum, Rhizopus nigricans, and Botrytis sp., were also carried out. Sodium tetraphosphate appeared to be the most effective compound in preserving cherries and also had the greatest antimycotic effects in the in vitro studies. A 10% concentration, when applied as a dip, inhibited fungal growth on fresh cherries for up to 30 days of storage at 1.1 C (34 F) and a relative humidity of 94%, whereas untreated controls showed fungal growth at 14 days. Following in order of effectiveness were sodium hexametaphosphate, sodium tripolyphosphate, and tetrasodium pyrophosphate.

Antifungal Agents↗

Hanseniaspora nodinigri, a new yeast species found in black knots (Dibotryon morbosum) of Prunus virginiana.

The new yeast species Hanseniaspora nodinigri is described to accommodate members of the genus Hanseniaspora that are unable to assimilate glucono-sigma-lactone and isolated from stromatal tissue of black knots (Dobotryon morbosum) of chokecherry, Prunus virginiana. The newly described taxon shows much resemblance, by other criteria, to H. vineae van der Walt et Tscheuschner and H. osmophila (Niehaus) Phaff, Miller et Shifrine.

Ascomycota↗

Sour Cherry (Prunus cerasus L) Anthocyanins as Ingredients for Functional Foods.

In the recent years many studies on anthocyanins have revealed their strong antioxidant activity and their possible use as chemotherapeutics. The finding that sour cherries (Prunus cerasus L) (also called tart cherries) contain high levels of anthocyanins that possess strong antioxidant and anti-inflammatory properties has attracted much attention to this species. Here we report the preliminary results of the induction of anthocyanin biosynthesis in sour cherry callus cell cultures. The evaluation and characterization of the in vitro produced pigments are compared to those of the anthocyanins found in vivo in fruits of several sour cherry cultivars. Interestingly, the anthocyanin profiles found in whole fruit extracts were similar in all tested genotypes but were different with respect to the callus extract. The evaluation of antioxidant activity, performed by ORAC and TEAC assays, revealed a relatively high antioxidant capacity for the fruit extracts (from 1145 to 2592 $\mu $ mol TE/100 g FW) and a lower one for the callus extract (688 $\mu $ mol TE/100 g FW).

Journal Article↗

Biochemical characterization of Pru a 2, a 23-kD thaumatin-like protein representing a potential major allergen in cherry (Prunus avium).

BACKGROUND: The prevalence of allergy to fruits and vegetables increased with pollinosis over the last 10 years. So far, clusters of hypersensitivity have been established and corroborated by the molecular characterization of individual cross-reacting allergens. Several case studies demonstrated the existence of allergic reactions to fruits of the subfamily Prunoideae (apricots, cherries, plums and peaches). Here, we present the characterization of a major allergen in cherry. METHODS: Characterization was performed using IgE immunoblotting and immunoblot inhibition, N-terminal sequencing, mass spectroscopy analysis and PCR-based cDNA cloning. RESULTS: A 23-kD protein was identified as IgE-binding component. As all cherry-extract-reactive sera displayed IgE-binding to this band, it was designated a major allergen from Prunus avium (Pru a 2). Sequencing the corresponding cDNA identified Pru a 2 as a thaumatin-like protein belonging to the group 5 of pathogenesis-related proteins. CONCLUSIONS: A thaumatin-like protein in cherry has been identified as a major allergen (Pru a 2). Homologous proteins from the thaumatin family share sequence similarities and should therefore be checked for the capability to elicit an IgE-mediated allergic reaction.

Allergens↗

Biological activity and structural characterization of alkaline-soluble polysaccharides from the kernels of Prunus mume Sieb. et Zacc.

The fruits of Prunus mume Sieb. et Zacc. (Japanese name, ume) have been used as a traditional drug and health food. In order to study the active components of P. mume, the polysaccharide fractions were extracted with cold water, hot water and aqueous sodium hydroxide from the kernels of P. mume. We found that some of the polysaccharide fractions exhibited various types of biological activities such as mitogenesis, activation of the alternative pathway of complement and activation of clot formation in human plasma. A polysaccharide, P-1, obtained from the cold 0.5 M NaOH extract was purified by ion-exchange chromatography and gel-filtration, P-1 contained 62.0% neutral sugar as glucose and 38.4% uronic acid (as galacturonic acid), and was free from protein. The neutral sugars of P-1 were arabinose, xylose, rhamnose and galactose in a molar ratio of 9.4:3.4:1.1:1.0, following analysis by gas-liquid chromatography. In addition, galacturonic acid was identified by thin-layer chromatography. The molecular weight of P-1 was found to be more than 2,000,000 by gel-filtration on Toyopearl HW 65F. P-1 showed mitogenic activity towards spleen cells of both C3H/HeN and C3H/HeJ, suggesting that it was free from bacterial endotoxic lipopolysaccharides.

Animals↗

Structural analysis of alkaline-soluble polysaccharide, P-1, from the kernels of Prunus mume Sieb. et Zucc.

A polysaccharide fraction extracted with cold 0.5 M NaOH from the kernels of Prunus mume exhibited some biological activities. A polysaccharide, P-1, was purified from the 0.5M NaOH extract by ion-exchange chromatography and gel-filtration. The results of the structural analysis of P-1 to determine the relationship between the activities and the structure are described in this paper. In the mild acid hydrolysis of P-1, the nondialyzable hydrolysate (I-3) believed to be its core portion was obtained. The yield of I-3 was 26.0% and contained 59.8% uronic acid as galacturonic acid (GalA). The neutral sugars of I-3 were composed of rhamnose, xylose and galactose in a molar ratio of 1.0:3.4:0.3 following analysis by gas-liquid chromatography. The molecular weight of I-3 was estimated to be ca. 14000 by gel-filtration on Toyopearl HW55F. I-3 exhibited the mitogenic activity toward spleen cells as well as P-1. These facts appeared to confirm that I-3 was the core part of P-1 and important for its biological activity. I-3 was successfully reduced by the Taylor and Conrad method to avoid so much repetition. Methylation analysis of the reduced hydrolysate by gas-liquid chromatography and gas chromatography-mass spectroscopy showed that the ratio of 1,4-linked galactopyranosyl and 1,3,4-linked galactopyranosyl residues were significantly increased in comparison with native I-3. These results suggested that I-3 was composed of 1,4- and 1,3,4-linked galacturonic acid residues in the main chain.

Animals↗

The screening of Chinese crude drugs for Ca2+ antagonist activity: identification of active principles from the aerial part of Pogostemon cablin and the fruits of Prunus mume.

Hot aqueous extracts of 134 Chinese crude drugs were subjected to screening for inhibitory activity on K+ contracture of guinea pig taenia coli, and significant activity was observed in 17 crude drugs. Chemical investigations of two crude drugs, Kakko and Ubai, which originate from Pogostemon cablin and Prunus mume, respectively, were undertaken, and patchouli alcohol (I) and 5-(hydroxymethyl)-2-furaldehyde (II) were identified as their active principles, respectively.

Animals↗

DNA fingerprinting study on the intraspecific variation and the origin of Prunus yedoensis (Someiyoshino).

In order to investigate the intraspecific variation of Prunus yedoensis (Someiyoshino) and interspecific relationship among P. yedoensis, P. lannesiana (Oshimazakura) and P. pendula (Edohigan), DNA fingerprinting study was conducted by using two different kinds of probes, M13 repeat sequence and (GACA)4 synthetic oligonucleotide. In this study, 68 plants of P. yedoensis grown in 46 prefectures in Japan were investigated. All the P. yedoensis individuals investigated showed the completely same banding pattern, indicating their clonal origin from a single plant. On the other hand, each of P. lannesiana and P. pendula individuals investigated showed a unique banding pattern, suggesting a considerable amount of genetic variation in these two species. About 90% of bands in DNA fingerprints of P. yedoensis were detected in either P. lannesiana or P. pendula. This result supports the hypothesis that P. yedoensis is an interspecific hybrid between P. lannesiana and P. pendula. From those results, it is concluded that P. yedoensis was produced only once through hybridization between P. lannesiana and P. pendula, and that this particular hybrid plant has been spread vegetatively all over Japan.

Base Sequence↗

Immunochemical characterization of alkaline-soluble polysaccharide, P-1, from the kernels of Prunus mume Sieb. et Zucc.

Polyclonal antibodies against P-1, a pectic polysaccharide fraction extracted with 0.5 M NaOH from the kernels of Prunus mume and consisted of arabino-galacturonan, and I-3, the partial acid (0.1 M trifluoroacetic acid) hydrolysate of P-1, were prepared in Japanese white rabbits. Competitive ELISA experiments strongly suggested that anti P-1 and anti I-3 antibodies were different but P-1 and I-3 cross-reacted with each other to recognize a partly similar epitope structure. The reactivities of polysaccharide fractions from the raw flesh of P. mume, and the kernels of apricot and peach extracted with either water or sodium hydroxide were examined using both antisera by the indirect competitive ELISA method. The polysaccharide fractions extracted with sodium hydroxide solutions had the reactivities but not those extracted with cold and hot water. These facts suggested that the similar structure of polysaccharides to P-1 was present in the flesh of P. mume and the kernels of apricot and peach. However, neither pectin of apple nor citrus had reactivity with each antiserum. P-1 would be different in chemical structure from a commercially available pectin, a water-soluble polysaccharide from apple and citrus.

Animals↗

Complete amino acid sequence determination of the major allergen of peach (Prunus persica) Pru p 1.

The major protein allergen of peach (Prunus persica), Pru p 1, has recently been identified as a lipid transfer protein (LTP). The complete primary structure of Pru p 1, obtained by direct amino acid sequence and liquid chromatography-mass spectrometry (LC-MS) analyses with the purified protein, is described here. The protein consists of 91 amino acids with a calculated molecular mass of 9178 Da. The amino acid sequence contains eight strictly conserved cysteines, as do all known LTPs, but secondary structure predictions failed to classify the peach 9 kDa protein as an 'all-alpha type', due to the high frequency of amino acids (nine prolines) disrupting alpha helices. Although the sequence similarity with maize LTP is only 63%, out of the 25 amino acids forming the inner surface of the tunnel-like hydrophobic cavity in maize ns-LTP 16 are identical and 7 similar in the peach homolog, supporting the hypothesis of a similar function.

Allergens↗

Cytological comparison of leaves and stems of Prunus avium L. shoots cultured on a solid medium with agar or gelrite.

An axillary proliferating clone of Prunus avium L. was subcultured every four weeks on solid MS medium with agar as the gelling agent. Vitrification (hyperhydricity) of shoots was induced in one four week cycle with the same medium except that agar was replaced by gelrite. During culture on the vitrifying medium, the water content of the shoots progressively increased with a parallel decrease in chlorophyll content. Cytological differences between the leaves and stems of the vitrified and normal shoots were detected by light and electron (both transmission and scanning) microscopy. Leaves of vitrified shoots were characterized by lower number of chloroplasts in the palisade parenchyma and by a defective cuticle. The stems of vitrified shoots had a less developed and lignifled xylem tissue, lacked sclerenchymatic areas and showed hypertrophy of the cortical parenchyma. More intense vacuolar activity with evaginations of the chloroplast envelope into the vacuole was noted in cells of vitrified leaves.

Agar↗

Relative concentration of plum pox virus in leaves and flowers of some Prunus species and cultivars.

The relative concentration of plum pox virus (PPV) in leaves and flowers of plum, damson, myrobalan, blackthorn, apricot and peach trees was determined by enzyme-linked immunosorbent assay (ELISA) and expressed as the lowest dilution with positive reaction. Significant differences in relative PPV concentration were found in leaves among individual Prunus species naturally or artificially infected with the virus. The highest relative PPV concentration was found in blackthorns (7.81 x 10(-4)), common plum and apricot (1.56 x 10(-3) for the both latters). Wild growing PPV-infected plums and blackthorns can be considered equally important source of sharka infection as PPV-susceptible cultivars of plums, apricots and peaches. High PPV concentration in flowers is of diagnostical value. High variability of relative PPV concentration was observed inside the species among individual cultivars. Susceptible cultivars were characteristic by high relative PPV concentration, e.g. apricot cvs. Vegama (9.8 x 10(-5)) and Velkopavlovická (1.95 x 10(-4)), and peach cvs. Maria Emilia (7.81 x 10(-4)) and Harbinger (1.56 x 10(-3)). On the other hand, cultivars resistant to PPV were characteristic by very low relative PPV concentration, e.g. apricot cv. Stark Early Orange (5 x 10(-2)) and peach cvs. Envoy (5 x 10(-2)) and Favorita Morettini (2.5 x 10(-2)). The highest relative PPV concentration was found in young trees newly infected with PPV.

Enzyme-Linked Immunosorbent Assay↗