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PubMed1991

Improvement of hyperglycemia and hyperlipemia in streptozotocin-diabetic rats by a methanolic extract of Prunus davidiana stems and its main component, prunin.

Blood glucose and total lipid levels in rats with streptozotocin-induced diabetes were determined after intraperitoneal administration of a methanolic extract of Prunus davidiana Fr. stems and its main component, prunin (= naringenin 7-O-beta-D-glucoside). From the data obtained it was concluded that intraperitoneal administration of the methanolic extract and prunin produced a significant hypoglycemic effect. Total blood lipids were also decreased by these substances. Thus, it is suggested that this methanolic extract contains one or more hypoglycemic principles including the main flavanone glycoside, prunin, which can significantly reduce the levels of blood glucose and total lipids in streptozotocin-diabetic rats.

Animals↗

PubMed1997

Identification of stylar RNases associated with gametophytic self-incompatibility in almond (Prunus dulcis).

Stylar proteins of 13 almond (Prunus dulcis) cultivars with known S-genotypes were surveyed by IEF and 2D-PAGE combined with immunoblot and N-terminal amino acid sequence analyses to identify S-RNases associated with gametophytic self-incompatibility (SI) in this plant species. RNase activities corresponding to Sa and Sb, two of the four S-alleles tested, were identified by IEF and RNase activity staining. The Sa-RNase band reacted with the anti-S4-serum prepared from Japanese pear (Pyrus serotina); no reaction with the antiserum was observed with the Sb-RNase band. When the Sa-RNase band was excised from an IEF gel stained for RNase activity, subjected to SDS-PAGE, and detected by immunoblotting, it appeared that this band consisted of a single protein that reacted with the anti-S4-serum with M(r) of about 28 kDa. With 2D-PAGE and silver staining of the stylar extracts, all four S-proteins could be successfully distinguished from each other in the highly basic zone of the gel. Although Sb-, Sc-, and Sd-proteins had roughly the same M(r) of about 30 kDa, the Sc-protein seemed to be slightly smaller than the Sb-protein and slightly larger than the Sd-protein. In 2D-PAGE profiles as well, the Sa-protein had M(r) of about 28 kDa, apparently smaller than the other three proteins. A bud sport, in which one of the two S-alleles of the original cultivar is impaired, was visualized as a loss of Sc-protein, which is consistent with the previous pollination study. All four S-proteins reacted with the anti-S4-serum, probably because of the differing conformations of these S-proteins in the IEF and 2D-PAGE gels. The Sa-protein in 2D-PAGE appeared to be identical to Sa-RNase in IEF; both had the same M(r) and were reactive with the anti-S4-serum. N-terminal amino acid sequence analysis of the four S-proteins revealed that they were highly homologous to each other and similar to the S-RNases of Malus, Pyrus, Scrophulariaceae, and Solanaceae. Taken together, RNases in the style are strongly suggested to be associated with the gametophytic SI of almond. This is the first report identifying and characterizing S-RNase in almond.

Amino Acid Sequence↗

PubMed2000

Isolation and characterization of a 60 kDa 2,4-D-binding protein from the shoot apices of peach trees (Prunus persica L.); it is a homologue of protein disulfide isomerase.

To obtain a candidate auxin-binding protein (ABP), a soluble 60 kDa protein was isolated from an extract of shoot apices of peach trees (Prunus persica L.) by affinity chromatography on a 2,4-dichlorophenoxyacetic acid (2,4-D)-linked Sepharose4B column. The 60 kDa polypeptide, designated Pp60, was purified as a single band on SDS-PAGE by column chromatography. Its dissociation constant (Kd) for [14C]-2,4-D was calculated to be 3.5 x 10(-5) M. The binding of Pp60 for [14C]-2,4-D was inhibited by naphthalene-1-acetic acid (NAA) and p-chlorophenoxyisobutyric acid (PCIB) as well as 2,4-D. Indole-3-acetic acid (IAA) had little effect on the binding. These results suggested that Pp60 is a protein that has an affinity for 2,4-D, NAA, and PCIB in vitro. The partial amino acid sequences of Pp60 showed high homology to those of protein disulfide isomerase (EC 5.3.4.1). Immunoblot analysis demonstrated that Pp60 exists ubiquitously in shoots and leaves. In fruit, expression of Pp60 is restricted at an early stage of development.

2,4-Dichlorophenoxyacetic Acid↗

PubMed1993

Leaf area and net photosynthesis during development of Prunus serotina seedlings.

We used the plastochron index to study the relationship between plant age, leaf age and development, and net photosynthesis of black cherry (Prunus serotina Ehrh.) seedlings. Leaf area and net photosynthesis were measured on all leaves >/= 75 mm of plants ranging in age from 7 to 20 plastochrons. Effects of plant developmental stage on leaf area and net photosynthesis were evaluated for leaves of differing age (horizontal series), leaves on plants of constant age (vertical series), and leaves of constant age (oblique series). Regression techniques were used to estimate leaf area from leaf blade dimensions. The best equations for predicting leaf area had R(2) values of 0.991-0.992 and used linear or logarithmic functions of both leaf length and width. Suitable, but less precise, equations with R(2) values of 0.946-0.962 were developed from either leaf length or leaf width. Leaf area development in black cherry seedlings was similar to that in other indeterminate species. Leaves of young plants reached full expansion at a lower leaf plastochron age than leaves of older plants. Maximum net photosynthesis per unit leaf area occurred 2-3 plastochrons before full leaf expansion. There was strong ontogenetic drift in net photosynthesis with leaf age; net photosynthesis decreased as plant age increased in leaves of the same plastochron age. Plots of the oblique series were particularly useful in providing information about interaction effects.

Journal Article↗

PubMedDate not supplied

Shade, leaf growth and crown development of Quercus rubra, Quercus velutina, Prunus serotina and Acer rubrum seedlings.

The study was conducted in an open field to determine the optimum irradiance for establishment and growth of two oak species and two major associated woody species. Half-sib seedlings of black cherry (Prunus serotina Ehrh.), red maple (Acer rubrum L.), northern red oak (Quercus rubra L.) and black oak (Q. velutina Lam.) were grown for two years under shade-cloth tents. Eight shade treatments (94, 70, 57, 45, 37, 27, 20 and 8% of full sunlight) with three replications each were used. Measurements were made on seedlings harvested at the end of the first and second growing seasons. In the second year, shading significantly decreased the number of leaves for all species except black cherry, but only significantly decreased leaf area in northern red oak. Shading significantly decreased average leaf size of red maple. Average leaf size of black cherry was largest in the intermediate shade treatments and decreased significantly with increased and decreased shade. Leaf weight/leaf area (mg cm(-2)) increased significantly in a quadratic pattern with decreasing shade for all four species. Leaf area ratio (cm(2) g(-1)) decreased significantly with decreasing shade for all species except red maple in the first year and black oak in the second year. Total branch development increased significantly with decreasing shade in red maple and northern red oak, whereas indeterminate branches increased significantly with decreasing shade only in black cherry, and short branches increased significantly with decreasing shade only in red maple.

Journal Article↗

PubMedDate not supplied

Effects of elevated CO(2) on growth and chloroplast proteins in Prunus avium.

To predict the future carbon sequestering capacity of trees, we need information about the possible acclimatory mechanisms of plant growth and photosynthesis in rising atmospheric CO(2) under a variety of environmental conditions. We have, therefore, studied the growth response of a tree species (Prunus avium L. Stella (wild cherry)) to elevated CO(2) and characterized the associated changes in photosynthetic machinery of the leaf tissue. Self-pollinated seedlings and mature cuttings (clones) from the same parent plant of P. avium were grown for two consecutive growing seasons (about 60 days each) in ambient CO(2) (350 micro mol mol(-1) CO(2)) or elevated CO(2) (700 micro mol mol(-1) CO(2)) with a high or low nutrient supply. The degree of acclimation of leaf biochemistry and growth response to elevated CO(2) was dependent on the plant material (seedling or mature cutting) and nutrient supply. There was little or no growth response to elevated CO(2) in seedlings or cuttings in the low nutrient supply treatments, whereas, in both seasons, there was a strongly positive growth response to elevated CO(2) in seedlings and cuttings in the high nutrient supply regimes, resulting in increases in the root/shoot ratio and in carbon allocation to the roots. In contrast, the protein content and activity of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco, EC 4.1.1.39) were down regulated in elevated CO(2). The loss of Rubisco on an area basis in plants in the elevated CO(2) treatments was compensated for at the canopy level by increased leaf area. The loss of Rubisco protein was accompanied by decreases in the contents of chlorophyll and the thylakoid membrane proteins D(1), D(2) and cytochrome f, which are involved in light harvesting and photo-electron transport. We conclude that, in the medium- to long-term, the initial stimulation of biomass production by elevated CO(2) may be increasingly offset by a lower photosynthetic capacity per unit leaf area in perennial plants.

Journal Article↗

PubMed1995

Arbuscular mycorrhizal induced changes to plant growth and root system morphology in Prunus cerasifera.

We compared root system morphogenesis of micropropogated transplants of Prunus cerasifera L. inoculated with either of the arbuscular mycorrhizal (AM) fungi Glomus mosseae or Glomus intraradices or with the ericoid mycorrhizal species Hymenoscyphus ericae. All plants were grown in sand culture, irrigated with a nutrient solution that included a soluble source of phosphorus, for 75 days after transplanting. Arbuscular mycorrhizal colonization increased both the survival and growth (by over 100%) of transplants compared with either uninoculated controls or transplants inoculated with H. ericae. Arbuscular mycorrhizal colonization increased root, stem and leaf weights, leaf area, root length and specific leaf area, and it decreased root length/leaf area ratio, root/shoot weight ratio and specific root length. Both uptake of phosphorus and its concentration in leaves were increased by AM infection, although the time course of the relationships between intensity of AM infection and P nutrition were complex and suggested a role for factors other than nutrition. The time course for the development of infection varied. It was most rapid with G. mosseae, but it was ultimately higher with G. intraradices. None of the treatments significantly affected the lengths of adventitious roots or the first-, second- or third-order laterals that developed from them. Arbuscular mycorrhizal colonization increased the intensity of branching in all root orders with the effect being most obvious on first-order lateral roots where the number of branches increased from under 100 to over 300 brances m(-1). As a result, although first-order laterals made up 55% of the root systems of control plants, the comparable value was 36% in AM-infected plants. In contrast, second-order laterals represented 25% of control root systems, but 50% of AM-colonized root systems. Glomus intraradices but not G. mosseae increased root diameter. Anatomical studies revealed no changes in the overall form of the root tip, although there were changes in the diameter of the root cap, cell numbers and cell size. Hymenoscyphus ericae increased the duration of the metaphase index. Both AM fungal treatments increased the concentrations of soluble proteins in root extracts and modified the protein profiles by the elimination and addition of protein bands detected by PAGE analysis. We conclude that AM fungal inoculation influenced processes in the root system at different levels, but not all effects were due to improved P nutrition or increased physiological age.

Journal Article↗

PubMed1996

Water stress and crop load effects on fruit fresh and dry weights in peach (Prunus persica).

Effects of water stress on fruit fresh and dry weights were investigated in peach trees, Prunus persica (L.) Batsch., with varying crop loads: light, moderate and heavy. In well-watered controls, tree water status was independent of crop load. In trees receiving reduced irrigation, the degree of water stress increased with increasing crop load. Water stress induced fruit fresh weight reductions at all crop loads. Fruit dry weight was not reduced by water stress in trees having light to moderate crop loads, indicating that the degree of water stress imposed did not affect the dry weight sink strength of fruit. Water-stressed trees with heavy crop loads had significantly reduced fruit dry weights, which were likely due to carbohydrate source limitations resulting from large crop carbon demands and water stress limitations on photosynthesis.

Journal Article↗

PubMed1996

Photoperiod effect on bud burst in Prunus is phase dependent: significance for early photosynthetic development.

A 16-h photoperiod stimulated bud burst in mature rooted cuttings of Prunus avium cv. Stella, but not in selfed Stella seedlings. However, in a 12-h photoperiod, bud burst occurred earlier in the seedlings than in the mature cuttings. In the 12-h photoperiod, production of polypeptides involved in carbon dioxide fixation and photosynthetic electron transport was higher in seedlings than in mature cuttings, whereas in the 16-h photoperiod, shoot development and polypeptide production were similar in seedlings and mature cuttings. In both photoperiods, the amount of large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase that was associated with thylakoid membranes in young leaves was higher in seedlings than in mature cuttings. Transcript levels of rbcL mRNA were influenced by photoperiod in mature cuttings but not in seedlings. In seedlings, early bud burst and development of the light harvesting apparatus would be an advantage at the start of the growing season, before the mature tree canopy reduces irradiances near the woodland floor.

Journal Article↗

PubMed1997

Crop load and water stress effects on daily stem growth in peach (Prunus persica).

We investigated crop load and water stress effects on diurnal stem extension growth of field-grown peach (Prunus persica (L.) Batsch) trees. Neither the presence of fruit nor reduced irrigation significantly altered the timing of diurnal fluctuations in stem growth rate. Stems with subtending fruit had significantly reduced growth compared to stems with no subtending fruit. Crop load had no significant effect on relative stem extension rates and the majority of the reduction in absolute growth was the result of a smaller zone of elongation in fruit-bearing stems than in stems with no subtending fruit. Fruit removal did not increase growth rates within 24 h. When irrigation was reduced, the length of the stem elongation zone and total daily stem growth were significantly decreased relative to well-irrigated controls and the decreases were highly correlated with stem water potential. Compared with well-irrigated controls, relative stem extension rates of water-stressed trees were reduced at several times during the 24-h period, but the degree of reduction was not proportional to the difference in stem water potentials between the treatments.

Journal Article↗

PubMed1998

Storage of foliar-absorbed nitrogen and remobilization for spring growth in young nectarine (Prunus persica var. nectarina) trees.

The effectiveness of spraying foliage with urea to provide nitrogen (N) to augment the seasonal internal cycling of N in young nectarine trees (Prunus persica (L.) Batsch var. nectarina (Ait. f. Maxim.), cv. Stark Red Gold) was studied. One-year-old trees were grown with contrasting N supplies during the summer and foliage was sprayed with a 2% urea solution labeled with (15)N just before leaf senescence started. After leaf abscission had finished, the trees were repotted in sand and given no further N. Remobilization of both labeled and unlabeled N for leaf growth the following spring was quantified. Leaves absorbed between 58 and 69% of the (15)N intercepted by the canopy irrespective of tree N status. During leaf senescence, the majority of (15)N was withdrawn from the leaves into the shoot and roots. Remobilization of (15)N the following spring was also unaffected by tree N status. About 38-46% of (15)N in the trees was recovered in the new growth. More unlabeled N (derived from root uptake) was remobilized for leaf growth in the spring than was withdrawn from leaves during canopy senescence the previous autumn. Therefore, soil-applied N augmented N storage pools directly, and contributed more to N remobilization the following spring than did foliar-absorbed (15)N.

Journal Article↗

PubMed1989

Selection for salt and drought tolerance in protoplast- and explant-derived tissue cultures of Colt cherry (Prunus avium x pseudocerasus).

Colt cherry (Prunus avium x pseudocerasus) callus cultures were derived from leaf protoplasts, protoplasts of root cell suspension cultures, or by direct culture of leaf and root tissues. Survival of calli cultured on basal proliferation medium containing 25, 50, 100 or 200 mN (millinormal) NaCl, Na(2)SO(4) or KCl, or iso-osmotic (with NaCl) concentrations of mannitol ranged from 1 to 15%. After six transfers on the same medium, surviving cell lines were subjected to three cycles of direct recurrent selection; i.e., in each cycle, they were cultured alternately on basal proliferation medium, and on basal proliferation medium supplemented with NaCl, KCl, Na(2)SO(4) or mannitol. Salt- or mannitol-tolerant cell lines selected in this way had smaller cells than unselected cell lines, and they grew more rapidly and had higher callus and cell survival rates than unselected cell lines when cultured in the presence of salt or mannitol. Cells lines selected for tolerance to one agent (sodium salt, potassium salt or mannitol) showed minimal tolerance to another agent. However, when plants were regenerated from salt- or mannitol-tolerant callus and new cultures derived from them, the new cultures showed tolerance to all of the salts and mannitol. Plant regeneration from the new cultures was not achieved under the conditions that led to the regeneration of the parent plants from callus.

Journal Article↗

PubMed1995

Prunus necrotic ringspot ilarvirus: nucleotide sequence of RNA3 and the relationship to other ilarviruses based on coat protein comparison.

The RNA3 of prunus necrotic ringspot ilarvirus (PNRSV) has been cloned and its entire sequence determined. The RNA3 consists of 1943 nucleotides (nt) and possesses two large open reading frames (ORFs) separated by an intergenic region of 74 nt. The 5' proximal ORF is 855 nt in length and codes for a protein of molecular mass 31.4 kDa which has homologies with the putative movement protein of other members of the Bromoviridae. The 3' proximal ORF of 675 nt is the cistron for the coat protein (CP) and has a predicted molecular mass of 24.9 kDa. The sequence of the 3' non-coding region (NCR) of PNRSV RNA3 showed a high degree of similarity with those of tobacco streak virus (TSV), prune dwarf virus (PDV), apple mosaic virus (ApMV) and also alfalfa mosaic virus (AIMV). In addition it contained potential stem-loop structures with interspersed AUGC motifs characteristic for ilar- and alfamoviruses. This conserved primary and secondary structure in all 3' NCRs may be responsible for the interaction with homologous and heterologous CPs and subsequent activation of genome replication. The CP gene of an ApMV isolate (ApMV-G) of 657 nt has also been cloned and sequenced. Although ApMV and PNRSV have a distant serological relationship, the deduced amino acid sequences of their CPs have an identity of only 51.8%. The N termini of PNRSV and ApMV CPs have in common a zinc-finger motif and the potential to form an amphipathic helix.

Amino Acid Sequence↗

PubMed1997

Replication of alfalfa mosaic virus RNA 3 with movement and coat protein genes replaced by corresponding genes of Prunus necrotic ringspot ilarvirus.

Alfalfa mosaic virus (AMV) and Prunus necrotic ringspot virus (PNRSV) are tripartite positive-strand RNA plant viruses that encode functionally similar translation products. Although the two viruses are phylogenetically closely related, they infect a very different range of natural hosts. The coat protein (CP) gene, the movement protein (MP) gene or both genes in AMV RNA 3 were replaced by the corresponding genes of PNRSV. The chimeric viruses were tested for heterologous encapsidation, replication in protoplasts from plants transformed with AMV replicase genes P1 and P2 (P12 plants) and for cell-to-cell transport in P12 plants. The chimeric viruses exhibited basic competence for encapsidation and replication in P12 protoplasts and for a low level of cell-to-cell movement in P12 plants. The potential involvement of the MP gene in determining host specificity in ilarviruses is discussed.

Alfalfa mosaic virus↗

PubMed1998

Virulence and molecular polymorphism of Prunus necrotic ringspot virus isolates.

Prunus necrotic ringspot virus (PNRSV) occurs as numerous strains or isolates that vary widely in their pathogenic, biophysical and serological properties. Prior attempts to distinguish pathotypes based upon physical properties have not been successful; our approach was to examine the molecular properties that may distinguish these isolates. The nucleic acid sequence was determined from 1.65 kbp RT-PCR products derived from RNA 3 of seven distinct isolates of PNRSV that differ serologically and in pathology on sweet cherry. Sequence comparisons of ORF 3a (putative movement protein) and ORF 3b (coat protein) revealed single nucleotide and amino acid differences with strong correlations to serology and symptom types (pathotypes). Sequence differences between serotypes and pathotypes were also reflected in the overall phylogenetic relationships between the isolates.

Amino Acid Sequence↗

PubMed2001

Recognition of cis-acting sequences in RNA 3 of Prunus necrotic ringspot virus by the replicase of Alfalfa mosaic virus.

Alfalfa mosaic virus (AMV) and Prunus necrotic ringspot virus (PNRSV) belong to the genera ALFAMOVIRUS: and ILARVIRUS:, respectively, of the family BROMOVIRIDAE: Initiation of infection by AMV and PNRSV requires binding of a few molecules of coat protein (CP) to the 3' termini of the inoculum RNAs and the CPs of the two viruses are interchangeable in this early step of the replication cycle. CIS:-acting sequences in PNRSV RNA 3 that are recognized by the AMV replicase were studied in in vitro replicase assays and by inoculation of AMV-PNRSV RNA 3 chimeras to tobacco plants and protoplasts transformed with the AMV replicase genes (P12 plants). The results showed that the AMV replicase recognized the promoter for minus-strand RNA synthesis in PNRSV RNA 3 but not the promoter for plus-strand RNA synthesis. A chimeric RNA with PNRSV movement protein and CP genes accumulated in tobacco, which is a non-host for PNRSV.

3' Untranslated Regions↗

PubMed2004

RNA-binding properties and mapping of the RNA-binding domain from the movement protein of Prunus necrotic ringspot virus.

The movement protein (MP) of Prunus necrotic ringspot virus (PNRSV) is involved in intercellular virus transport. In this study, putative RNA-binding properties of the PNRSV MP were studied. The PNRSV MP was produced in Escherichia coli using an expression vector. Electrophoretic mobility shift assays (EMSAs) using DIG-labelled riboprobes demonstrated that PNRSV MP bound ssRNA cooperatively without sequence specificity. Two different ribonucleoprotein complexes were found to be formed depending on the molar MP : PNRSV RNA ratio. The different responses of the complexes to urea treatment strongly suggested that they have different structural properties. Deletion mutagenesis followed by Northwestern analysis allowed location of a nucleic acid binding domain to aa 56-88. This 33 aa RNA-binding motif is the smallest region delineated among members of the family Bromoviridae for which RNA-binding properties have been demonstrated. This domain is highly conserved within all phylogenetic subgroups previously described for PNRSV isolates. Interestingly, the RNA-binding domain described here and the one described for Alfamovirus are located at the N terminus of their corresponding MPs, whereas similar domains previously characterized in members of the genera Bromovirus and Cucumovirus are present at the C terminus, strongly reflecting their corresponding phylogenetic relationships. The evolutionary implications of this observation are discussed.

Base Sequence↗

PubMed1992

Prunus serotina Amygdalin Hydrolase and Prunasin Hydrolase : Purification, N-Terminal Sequencing, and Antibody Production.

In black cherry (Prunus serotina Ehrh.) seed homogenates, amygdalin hydrolase (AH) participates with prunasin hydrolase (PH) and mandelonitrile lyase in the sequential degradation of (R)-amygdalin to HCN, benzaldehyde, and glucose. Four isozymes of AH (designated AH I, I', II, II') were purified from mature cherry seeds by concanavalin A-Sepharose 4B chromatography, ion-exchange chromatography, and chromatofocusing. All isozymes were monomeric glycoproteins with native molecular masses of 52 kD. They showed similar kinetic properties (pH optima, K(m), V(max)) but differed in their isoelectric points and N-terminal amino acid sequences. Analytical isoelectric focusing revealed the presence of subisozymes of each isozyme. The relative abundance of these isozymes and/or subisozymes varied from seed to seed. Three isozymes of PH (designated PH I, IIa, and IIb) were purified to apparent homogeneity by affinity, ion-exchange, and hydroxyapatite chromatography and by nondenaturing polyacrylamide gel electrophoresis. PH I and PH IIb are 68-kD monomeric glycoproteins, whereas PH IIa is dimeric (140 kD). The N-terminal sequences of all PH and AH isozymes showed considerable similarity. Polyclonal antisera raised in rabbits against deglycosylated AH I or a mixture of the three deglycosylated PH isozymes were not monospecific as judged by immunoblotting analysis, but also cross-reacted with the opposing glucosidase. Monospecific antisera deemed suitable for immunocytochemistry and screening of expression libraries were obtained by affinity chromatography. Each antiserum recognized all known isozymes of the specific glucosidase used as antigen.

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