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Crystal macropattern development in Prunus serotina (Rosaceae, Prunoideae) leaves.

BACKGROUND AND AIMS: Prunus, subgenus Padus, exhibits two completely different calcium oxalate crystal macropatterns in mature leaves. Foliar macropattern development has been described previously in P. virginiana, representing one version. Prunus serotina, in the group exhibiting the second macropattern, is described here. The goal was to describe developmental details for comparison with P. virginiana, and to extend the sparse current knowledge of crystal macropatterns. METHODS: Leaves at various developmental stages were removed from local trees and from herbarium specimens. Early leaf stages and freehand leaf and stem sections were mounted directly in aqueous glycerine; larger leaves were processed whole or in representative pieces in household bleach, dehydrated in alcohol/xylol, and mounted in Permount. Crystals were detected microscopically between crossed polarizers. KEY RESULTS: Bud scales have a dense druse population. Druses appear first at the stipule tip and proliferate basipetally but soon stop forming; growing stipules therefore have a declining density of druses. Druses appear at the tip of leaves <1 mm long, then proliferate basipetally in the midrib. Lamina druses appear in the distal marginal teeth of leaves 3 cm long; from here they proliferate basipetally and towards midrib along major veins. In about two-thirds-grown leaves (6-9 cm length) druses are all adaxial to veins of most orders; a shift occurs then to formation of prisms, which appear first abaxial to, then all around, veins. Mature leaves have virtually all prisms encrusting all major veins, more sparsely along smaller minor veins. Late season leaves form epitactic crystals on existing prismatics. CONCLUSIONS: The developing and mature macropattern of P. serotina is almost the reverse of the pattern described previously in P. virginiana, and shows that two closely related species can develop radically different modes of crystallization. The few detailed macropattern studies to date reveal striking variations that indicate a new level of organization that must be integrated with the anatomical, physiological and molecular approaches that have been dominant so far.

Calcium Oxalate↗

Isolation and quantitation of amygdalin in Apricot-kernel and Prunus Tomentosa Thunb. by HPLC with solid-phase extraction.

Apricot-kernel and Prunus Tomentosa Thunb. are traditional Chinese herb medicines that contain amygdalin as their major effective ingredient. In this report, three methods for the extraction of amygdalin from the medicinal materials are compared: ultrasonic extraction by methanol, Soxhlet extraction by methanol, and reflux extraction by water. The results show that reflux extraction water containing 0.1% citric acid is the best option. The optimal reflux is 2.5 h and water bath temperature is 60 degrees C. The solid-phase extraction method using C18 and multiwalled carbon nanotube as adsorbents is established the pretreatment of reflux extract, and the result shows that the two adsorbents have greater adsorptive capacity for amygdalin and good separation effect. In order to quantitate amygdalin in Apricot-kernel and Prunus Tomentosa Thunb., a reversed-phase high-performance liquid chromatography method using methanol-water (15:85, for 30 min and pure methanol after 30 min) as mobile phase is developed and a good result is obtained.

Amygdalin↗

Ethylene-responsive genes are differentially regulated during abscission, organ senescence and wounding in peach (Prunus persica).

Ethylene-responsive genes from peach (Prunus persica, L. Batsch) were isolated by differential screening of a cDNA library constructed from abscission zones in which cell separation had been evoked by treatment with the ethylene analogue propylene. DNA and deduced protein sequences of four selected clones, termed Prunus persica Abscission zone (PpAz), revealed homology to thaumatin-like proteins (PpAz8 and PpAz44), to proteins belonging to the PR4 class of pathogenesis-related (PR) proteins (PpAz89), and to fungal and plant beta-D-xylosidases (PpAz152). Expression analyses conducted on embrioctomized and CEPA-treated fruitlets as well as on fruit explants have shown that PpAz8, PpAz44 and PpAz89 are preferentially transcribed in the cells of the fruit abscission zone rather than in the non-zone tissues. The PpAz152 transcript showed a different accumulation pattern being consistently and promptly induced by wounding and only slightly stimulated by propylene. By contrast, a complex pattern of transcript accumulation was found for the four genes in response to the wounding of leaves and during organ development and senescence. Based on this evidence, the existence of multiple regulatory pathways underlying the differential expression of the four PpAz genes in the different tissues and physiological processes is hypothesized.

Alkenes↗

Self-incompatibility (S) locus region of the mutated S6-haplotype of sour cherry (Prunus cerasus) contains a functional pollen S allele and a non-functional pistil S allele.

This study characterizes the S6m-haplotype, a mutated S6-haplotype with an altered HindIII cut site, of sour cherry (Prunus cerasus). Inheritance and pollination studies of S-haplotypes from reciprocal crosses between 'Erdi Botermo' (EB; S4S6mSa) and 'Rheinische Schattenmorelle' (RS; S6SaSbSc) revealed that the S6m-haplotype conferred unilateral incompatibility with a non-functional pistil component and a functional pollen component. Expression analyses of S6-RNase and SFB6, a candidate gene for pollen-S, in the S6m-haplotype showed that SFB6 was transcribed in EB pollen, but S6-RNase was not transcribed in EB styles. These results were consistent with data from the inheritance and pollination studies. Inverse PCR for the flanking regions of S6-RNase in the S6- and S6m-haplotypes revealed an approximately 2600 bp insertion present at approximately 800 bp upstream of the S6-RNase in the S6m-haplotype, which is responsible for the alternation of the HindIII cut site and a possible cause of inhibition of the transcription of S6-RNase. SFB6 was present downstream of S6-RNase in both the S6- and S6m-haplotypes and expressed in the same way, supporting the idea that SFB is a good candidate for pollen-S in Prunus.

Alleles↗

The role of polar auxin transport through pedicels of Prunus avium L. in relation to fruit development and retention.

It was investigated whether premature fruit abscission in Prunus avium L. was triggered by a reduction in polar auxin transport (PAT). The capacity of pedicels to transport tritiated IAA ([3H]-IAA) via the PAT pathway was measured at intervals throughout flower and fruit development. The extent of passive diffusion, assessed by concurrent applications of [14C]-benzoic acid ([14C]-BA), was negligible. Transported radioactivity recovered from agar blocks eluted at the same retention time as authentic [3H]-IAA during HPLC fractionation. The capacity for PAT was already high 7 d before anthesis and increased further following the fertilization of flowers at anthesis. PAT intensity was greatest immediately following fertilization and at the beginning of the cell expansion phase of fruit growth; the transport intensity in fruitlets destined to abscind was negligible. The amount of endogenous IAA moving through the PAT pathway was greatest during the first 3 weeks after fertilization and was again high at the beginning of the fruit expansion stage. IAA export in the phloem increased following fertilization then declined below detectable levels. ABA export in the phloem increased markedly during stone formation and at the onset of fruit expansion. TIBA applied to pedicels of fruit in situ promoted fruitlet abscission in 2000 but not in 2001, despite PAT capacity being reduced by over 98% in the treated pedicels. The application of TIBA to pedicels did not affect fruit expansion. The role of PAT and IAA in relation to the development and retention of Prunus avium fruit is discussed.

Abscisic Acid↗

Simulating genotypic variation of fruit quality in an advanced peach x Prunus davidiana cross.

Ecophysiological models are increasingly expected to describe genotypic variation within breeding populations. Accordingly, the ability of an ecophysiological model of peach to explain variation in fruit quality among 100 genotypes of a second backcross progeny derived from a clone of wild peach (Prunus davidiana) crossed with two commercial nectarine (Prunus persica) varieties was explored. Experimental measurements were carried out to calibrate the model for each genotype. The predictive quality of the model was tested on several independent datasets. The genotypic variation in dry and fresh growth of the fruit and the stone were effectively described by the model. Prediction of the amount of total sugar in flesh at maturity was accurate, whereas prediction of flesh dry matter content and total sugar concentration was suitable but less accurate. This approach and the results have allowed physiological processes to be ranked according to their contribution to the variation in fruit quality between genotypes. Fruit growth demand and the hydraulic conductance in the fruit were the main processes that explained the fruit quality variation. Shortcomings and further potential uses of the model are discussed.

Agriculture↗

A pollen-expressed gene for a novel protein with an F-box motif that is very tightly linked to a gene for S-RNase in two species of cherry, Prunus cerasus and P. avium.

This study describes a novel F-box protein gene in the S-locus of sour cherry (Prunus cerasus) and sweet cherry (P. avium). The gene showed an S-haplotype-specific sequence polymorphism and the expression was specific to pollen. Genomic DNA blot analysis of eight sweet cherry cultivars with the probe for the F-box protein gene under low stringency conditions yielded RFLP bands specific to the S-haplotypes of each cultivar. We discuss the possibility of the gene for the F-box protein being a candidate for the male determinant of gametophytic self-incompatibility in PRUNUS:

Amino Acid Sequence↗

High-throughput targeted SSR marker development in peach (Prunus persica).

Simple sequence repeats (SSRs) have proven to be highly polymorphic, easily reproducible, codominant markers. However, developing an SSR map is very time consuming and expensive, and most SSRs are not specifically linked to gene loci of immediate interest. The ideal situation would be to combine a high-throughput, relatively inexpensive mapping technique with rapid identification of SSR loci in mapped regions of interest. For this reason, we coupled the high-throughput technique of AFLP mapping with subsequent direct targeting of SSRs identified in AFLP-marked regions of interest. This approach relied on the availability of peach bacterial artificial chromosome (BAC) library resources. We present examples of using this strategy to rapidly identify SSR loci tightly linked to two important, simply inherited traits in peach (Prunus persica (L.) Batsch): root-knot nematode resistance and control of the evergrowing trait. SSRs developed in this study were also tested for their transportability in other Prunus species and in apricots.

Alleles↗

Prunus spp. intoxication in ruminants: a case in a goat and diagnosis by identification of leaf fragments in rumen contents.

Prunus serotina Ehrh. (black cherry) intoxication was diagnosed on postmortem examination of a goat. The clinical signs were weakness, depression, seizure-like activity, and lateral recumbency. Natural cases of black cherry intoxication have not been reported in goats in the United States. In the absence of a history of access to black cherry or the ability to detect cyanide or cyanogenic glycosides in blood or tissues, black cherry intoxication may be diagnosed in ruminants by the identification of black cherry leaves in rumen contents. Three distinctive features facilitate identification of black cherry leaves or leaf fragments: 1) a pair of small glands that protrude from the sides of the petiole just below the base of the blade, 2) incurved, gland-tipped (callous) teeth along the margins of the leaf, and 3) a band of hairs to each side of the lower half of the midvein on the surface of the leaf. Shape of the marginal teeth, presence or absence of glands at the tips of these teeth, the morphology of these glands, and presence or absence of petiolar glands and their morphology may allow identification and differentiation of small fragments of leaves from the 6 most important cyanogenic Prunus spp. in eastern North America: black cherry, Carolina laurel cherry, peach, English laurel cherry, choke cherry, and fire cherry.

Animals↗

Mapping with a few plants: using selective mapping for microsatellite saturation of the Prunus reference map.

The concept of selective (or bin) mapping is used here for the first time, using as an example the Prunus reference map constructed with an almond x peach F2 population. On the basis of this map, a set of six plants that jointly defined 65 possible different genotypes for the codominant markers mapped on it was selected. Sixty-three of these joint genotypes corresponded to a single chromosomal region (a bin) of the Prunus genome, and the two remaining corresponded to two bins each. The 67 bins defined by these six plants had a 7.8-cM average length and a maximum individual length of 24.7 cM. Using a unit of analysis composed of these six plants, their F1 hybrid parent, and one of the parents of the hybrid, we mapped 264 microsatellite (or simple-sequence repeat, SSR) markers from 401 different microsatellite primer pairs. Bin mapping proved to be a fast and economic strategy that could be used for further map saturation, the addition of valuable markers (such as those based on microsatellites or ESTs), and giving a wider scope to, and a more efficient use of, reference mapping populations.

Chromosome Mapping↗

Flavonoids from Prunus serotina Ehrh.

In the course of chemotaxonomic study of the genus Prunus, seven flavonol glycosides were isolated from the leaves of Prunus serotina Ehrh., characterized by UV and NMR spectroscopy, and identified finally as three quercetin monosides: hyperoside, avicularin, reynoutrin, three quercetin biosides: 3-O-(6"-O-alpha-L-rhamnopyranosyl)-beta-D-glucopyranoside, 3-O-(2"-O-alpha-L-rhamnopyranosyl)-beta-D-glucopyranoside and 3-O-(2"-O-alpha-L-rhamnopyranosyl)-beta-D-galactopyranoside as well isorhamnetin 3-O-(6"-O-alpha-L-rhamnopyranosyl)-beta-D-glucopyranoside. The presence of determined flavonoids in the flowers was confirmed by TLC.

Chromatography, Thin Layer↗

A highly conserved alpha-tubulin sequence from Prunus amygdalus.

The sequence of an alpha-tubulin from Prunus amygdalus has been obtained by cDNA cloning. When this sequence is compared to that of the Tub alpha 1 gene from maize it shows a very high degree of similarity, much higher than any of the alpha-tubulin sequences reported so far from plants. The expression of this gene is high in the stages of seed development where a high divisional activity is present. It is preferentially expressed in the radicular tissues as it is gene Tub alpha 1 in maize. Southern analysis indicates that this gene may form a subfamily of alpha-tubulin genes having similar sequence and tissue specificity and existing at least in maize and in Prunus.

Amino Acid Sequence↗

Candidate genes and QTLs for sugar and organic acid content in peach [ Prunus persica (L.) Batsch].

The identification of genes involved in variation of peach fruit quality would assist breeders in creating new cultivars with improved fruit quality. Major genes and quantitative trait loci (QTLs) for physical and chemical components of fruit quality have already been detected, based on the peach [ Prunus persica (L.) Batsch] cv. Ferjalou Jalousia((R)) (low-acid peach) x cv. Fantasia (normally-acid nectarine) F(2) intraspecific cross. Our aim was to associate these QTLs to structural genes using a candidate gene/QTL approach. Eighteen cDNAs encoding key proteins in soluble sugar and organic acid metabolic pathways as well as in cell expansion were isolated from peach fruit. A single-strand conformation polymorphism strategy based on specific cDNA-based primers was used to map the corresponding genes. Since no polymorphism could be detected in the Ferjalou Jalousia((R)) x Fantasia population, gene mapping was performed on the almond [ Prunus amygdalus ( P. dulcis)] cv. Texas x peach cv. Earlygold F(2) interspecific cross from which a saturated map was available. Twelve candidate genes were assigned to four linkage groups of the peach genome. In a second step, the previous QTL detection was enhanced by integrating anchor loci between the Ferjalou Jalousia((R)) x Fantasia and Texas x Earlygold maps and data from a third year of trait assessment on the Ferjalou Jalousia((R)) x Fantasia population. Comparative mapping allowed us to detect a candidate gene/QTL co-location. It involved a cDNA encoding a vacuolar H(+)-pyrophosphatase ( PRUpe;Vp2) that energises solute accumulation, and QTLs for sucrose and soluble solid content. This preliminary result may be the first step in the future development of marker-assisted selection for peach fruit sucrose and soluble solid content.

Journal Article↗

Ecophysiological and morphological responses to shade and drought in two contrasting ecotypes of Prunus serotina.

Photosynthesis (A), water relations and stomatal reactivity during drought, and leaf morphology were evaluated on 2-year-old, sun- and shade-grown Prunus serotina Ehrh. seedlings of a mesic Pennsylvania seed source and a more xeric Wisconsin source. Wisconsin plants maintained higher A and leaf conductance (g(wv)) than Pennsylvania plants during the entire drought under sun conditions, and during the mid stages of drought under shade conditions. Compared to shade plants, sun plants of both sources exhibited a more rapid decrease in A or % A(max) with decreasing leaf water potential (Psi). Tissue water relations parameters were generally not significantly different between seed sources. However, osmotic potentials were lower in sun than shade plants under well-watered conditions. Following drought, shade plants, but not sun plants, exhibited significant osmotic adjustment. Sun leaves had greater thickness, specific mass, area and stomatal density and lower guard cell length than shade leaves in one or both sources. Wisconsin sun leaves were seemingly more xerophytic with greater thickness, specific mass, and guard cell length than Pennsylvania sun leaves. No source differences in leaf structure were exhibited in shade plants. Stomatal reactivity to sun-shade cycles was similar between ecotypes. However, well-watered and droughted plants differed in stomatal reactivity within and between multiple sun-shade cycles. The observed ecotypic and phenotypic variations in ecophysiology and morphology are consistent with the ability of Prunus serotina to survive in greatly contrasting environments.

Journal Article↗

Comparison of drought resistance among Prunus species from divergent habitats.

Root and shoot characteristics related to drought resistance were compared among cultivated peach (Prunus persica (L.) Batsch.), P. andersonii (Nevada Desert almond), P. besseyi (western sand cherry), P. maritima (beach plum), P. subcordata (Sierra or Pacific plum), and P. tomentosa (Nanking cherry). In all species, shoot characteristics were more closely associated with drought adaptation than root characteristics. The most xeric species, P. andersonii, had the lowest specific leaf area, smallest leaves, highest stomatal conductance (before stress), highest rate of carbon assimilation (A), high root length/leaf area and root weight/leaf area ratios, and the highest leaf nitrogen content on an area basis. Root hydraulic conductivity was similar for all species, indicating a lack of importance of this parameter for drought resistance. During a 5-7 day drought, water use efficiency (WUE) increased as shoot water potentials (Psi) declined to -3.0 to -4.0 MPa for the xeric P. andersonii and P. subcordata, whereas after an initial increase, WUE decreased with declining Psi in the -1.5 to -3.0 MPa range for the more mesic P. maritima, P. persica and P. tomentosa as a result of non-stomatal limitations to A. Carbon assimilation rate decreased linearly with Psi during drought in all species, but the Psi at which A reached zero was not associated with drought adaptation. We conclude that the variation in leaf characteristics among Prunus species could be exploited to improve the drought resistance of commercial cultivars.

Journal Article↗

Sodium and chloride distribution in salt-stressed Prunus salicina, a deciduous tree species.

Measurements were made over four growing seasons of the Na(+) and Cl(-) content of leaves and woody tissues (twigs, branches, trunk and roots) of mature, fruit-bearing Prunus salicina Lindl. (on Marianna 2624 rootstock) trees irrigated during the growing season with water containing 3, 14 or 28 mM salt (2/1 molar ratio of NaCl and CaCl(2)). At the beginning of the study, the trees were 19 years old. Woody tissues of trees irrigated with water containing 14 or 28 mM salt accumulated Na(+) and Cl(-). Leaves of trees irrigated with water containing 14 or 28 mM salt accumulated Cl(-), but not Na(+), unless they had visible symptoms of salt injury. X-Ray microanalysis of leaf mesophyll cells indicated some ability of the cells to sequester Cl(-) in the vacuole. The data demonstrate a capacity for ion compartmentation among tissues and cell organelles in mature Prunus salicina, which may explain the ability of the species to survive low levels of salinity for several years in the field.

Journal Article↗

Development, distribution, and characteristics of intrinsic, nonbacterial ice nuclei in prunus wood.

Ice nuclei active at approximately -2 degrees C and intrinsic to woody tissues of Prunus spp. were shown to have properties distinct from bacterial ice nuclei. Soaking 5-centimeter peach stem sections in water for 4 hours lowered the mean ice nucleation temperature to below -4 degrees C, nearly 2 degrees C lower than stems inoculated with ice nucleation-active Pseudomonas syringae strain B301D. Ice nucleation activity in peach was fully restored by air-drying woody stem sections for a few hours. The ice nuclei in woody tissue were inactivated between 40 and 50 degrees C, but unaffected by treatment with bacterial ice nucleation inhibitors (i.e. NaOCl, tartaric acid, Triton XQS-20), sulfhydryl reagents (i.e. p-hydroxymercuribenzoate and iodine) and Pronase. Ice nuclei could not be dislodged from stems by sonication and were shown to be equally distributed in peach bud and internodal stem tissue on a per unit mass basis; outer and inner stem tissues were also indistinguishable in ice nucleation activity. Development of ice nuclei in immature peach and sweet cherry stems did not occur until midsummer and their formation was essentially complete by late August. Once formed the ice nuclei intrinsic to woody stems were stable and unaffected by seasonal changes in growth. The apparent physiological function of the ice nuclei is discussed in relation to supercooling and mechanisms of cold hardiness in Prunus spp.

Journal Article↗

Immunochemical and biological properties of a mouse monoclonal antibody reactive to prunus necrotic ringspot ilarvirus.

A monoclonal antibody reacting with prunus necrotic ringspot ilarvirus was tested in immunochemical studies, neutralization of infectivity assays, and by immuno-electron microscopy. The antibody was able to detect the 27,000 Mr coat protein of prunus necrotic ringspot ilarvirus in western blots and also detected all polypeptide fragments generated after incubation of whole virus with proteolytic enzymes. In neutralization of infectivity studies, the antibody blocked virus infectivity, although it did not precipitate the antigen in agar gel Ouchterlony double diffusion tests. Immuno-electron microscopy confirmed that the antibody coats virions but does not cause clumping. The antibody may be a useful tool for investigating coat protein-dependent initiation of ilarvirus infection.

Animals↗