Phytochemical composition and pigment stability of Açai (Euterpe oleracea Mart.)

Anthocyanin and polyphenolic compounds present in a?ai (Euterpe oleracea Mart.) were determined and their respective contribution to the overall antioxidant capacity established. Color stability of a?ai anthocyanins against hydrogen peroxide (0 and 30 mmol/L) over a range of temperatures (10-30 degrees C) was also determined and compared to common anthocyanin sources. Additionally, stability in a model beverage system was evaluated in the presence of ascorbic acid and naturally occurring polyphenolic cofactors. Cyanidin 3-glucoside (1040 mg/L) was the predominant anthocyanin in a?ai and correlated to antioxidant content, while 16 other polyphenolics were detected from 4 to 212 mg/L. Red grape anthocyanins were most stable in the presence of hydrogen peroxide, while a?ai and pigments rich in acylated anthocyanins displayed lower color stability in a temperature-dependent manner. In the presence of ascorbic acid, acylated anthocyanin sources generally had increased color stability. A?ai was recognized for its functional properties for use in food and nutraceutical products.     

Absorption and biological activity of phytochemical-rich extracts from açai (Euterpe oleracea Mart.) pulp and oil in vitro

Polyphenolic extracts from various fruits and vegetables have been shown to exert growth inhibitory effects in cell culture studies. Whereas individual polyphenolic compounds have been extensively evaluated, understanding of the biological activity of polyphenolic extracts from natural sources is limited and critical to the understanding of their potential effects on the human body. This study investigated the absorption and antiproliferative effects of phytochemical extracts from acai pulp and a polyphenolic-enriched acai oil obtained from the fruit pulp of the acai berry ( Euterpe oleracea Mart.). Chemical composition, antioxidant properties, and polyphenolic absorption of phytochemical fractions in a Caco-2 monolayer were determined, along with their cytotoxicity in HT-29 human colon adenocarcinoma cells. Standardized extracts were characterized by their predominance of hydroxybenzoic acids, monomeric flavan-3-ols, and procyanidin dimers and trimers. Polyphenolic mixtures (0-12 microg of gallic acid equiv/mL) from both acai pulp and acai oil extracts inhibited cell proliferation by up to 90.7%, which was accompanied by an increase of up to 2.1-fold in reactive oxygen species. Absorption experiments using a Caco-2 intestinal cell monolayer demonstrated that phenolic acids such as p-hydroxybenzoic, vanillic, syringic, and ferulic acids, in the presence of DMSO, were readily transported from the apical to the basolateral side along with monomeric flavanols such as (+)-catechin and (-)-epicatechin. Results from this study provide further evidence for the bioactive properties of acai polyphenolics and offer new insight on their composition and cellular absorption.     

Açai (Euterpe oleracea Mart.) polyphenolics in their glycoside and aglycone forms induce apoptosis of HL-60 leukemia cells

The effects of a?ai polyphenolics on the antiproliferation and induction of apoptosis in HL-60 human leukemia cells were investigated. Interactions between anthocyanins and non-anthocyanin-polyphenolics in both their glycosidic and their aglycone forms were also investigated to determine additive or nonadditive responses. Polyphenolic fractions at 0.17-10.7 microM were found to reduce cell proliferation from 56 to 86% likely due to caspase-3 activation (apoptosis). Anthocyanin and polyphenolic fractions were nonadditive in their contribution to the cell antiproliferation activity. At equimolar concentrations, the glycosidic forms of phenolic acids and flavonoids induced a higher magnitude of change in cell parameters (proliferation and apoptosis) than their respective aglycone forms, while the opposite trend was observed for anthocyanin aglycones. This study demonstrated that a?ai offers a rich source of bioactive polyphenolics and confirmed the importance of investigating whole food systems when evaluating the potential health benefits of individual phytochemical compounds.     

Anthocyanin-rich açai (Euterpe oleracea Mart.) fruit pulp fractions attenuate inflammatory stress signaling in mouse brain BV-2 microglial cells

Age-related diseases of the brain compromise memory, learning, and movement and are directly linked with increases in oxidative stress and inflammation. Previous research has shown that supplementation with berries can modulate signaling in primary hippocampal neurons or BV-2 mouse microglial cells. Because of their high polyphenolic content, fruit pulp fractions of a?ai ( Euterpe oleracea Mart.) were explored for their protective effect on BV-2 mouse microglial cells. Freeze-dried a?ai pulp was fractionated using solvents with different polarities and analyzed using HPLC for major anthocyanins and other phenolics. Fractions extracted using methanol (MEOH) and ethanol (ETOH) were particularly rich in anthocyanins such as cyanidin, delphinidin, malvidin, pelargonidin, and peonidin, whereas the fraction extracted using acetone (ACE) was rich in other phenolics such as catechin, ferulic acid, quercetin, resveratrol, and synergic and vanillic acids. Studies were conducted to investigate the mitigating effects of a?ai pulp extracts on lipopolysaccharide (LPS, 100 ng/mL) induced oxidative stress and inflammation; treatment of BV-2 cells with acai fractions resulted in significant (p < 0.05) decreases in nitrite production, accompanied by a reduction in inducible nitric oxide synthase (iNOS) expression. The inhibition pattern was emulated with the ferulic acid content among the fractions. The protection of microglial cells by a?ai pulp extracts, particularly that of MEOH, ETOH, and ACE fractions, was also accompanied by a significant concentration-dependent reduction in cyclooxygenase-2 (COX-2), p38 mitogen-activated protein kinase (p38-MAPK), tumor necrosis factor-α (TNFα), and nuclear factor κB (NF-κB). The current study offers valuable insights into the protective effects of a?ai pulp fractions on brain cells, which could have implications for improved cognitive and motor functions.     

Polyphenolics from açaí ( Euterpe oleracea Mart.) and red muscadine grape (Vitis rotundifolia ) protect human umbilical vascular Endothelial cells (HUVEC) from glucose- and lipopolysaccharide (LPS)-induced inflammation and target microRNA-126

Endothelial anti-inflammatory effects of ac?ai? (Ac) and red muscadine grape (Gp) polyphenolics have not been extensively investigated. It was hypothesized that polyphenolics from Ac and Gp exert comparable protective effects in human vascular endothelial cells (HUVEC) upon inflammatory stress. Furthermore, this study investigated whether microRNAs relevant to endothelial function might be regulated by Ac and Gp. Results showed that Ac and Gp (5-20 mg gallic acid equivalent/L) protected HUVEC against glucose-induced oxidative stress and inflammation. Glucose-induced expression of interleukin-6 and -8 was down-regulated by Ac and Gp at mRNA and protein levels. Upon lipopolysaccharide (LPS; 1 μg/L)-induced inflammation, Ac and Gp inhibited gene expression of adhesion molecules and NF-κB activation to similar extents, although Gp was more effective in decreasing PECAM-1 and ICAM-1 protein. Of the screened microRNAs, only microRNA-126 expression was found to be modulated by Ac and Gp as the underlying mechanism to inhibit gene and protein expression of VCAM-1.     

Effects of polyphenolic compounds on tumor necrosis factor-α (TNF-α)-induced changes of adipokines and oxidative stress in 3T3-L1 adipocytes

Over the last few decades, obesity has become a global epidemic in both developed and developing countries. Recent studies have indicated that obesity is closely associated with chronic inflammation characterized by abnormal levels of adipocytokines and inflammatory cytokines in adipocytes. The aim of this work was to study the effects of 21 polyphenolic compounds on tumor necrosis factor-α (TNF-α)-induced changes of adipokines and oxidative stress in 3T3-L1 adipocytes. The results showed that p-coumaric acid, quercetin, and resveratrol have greater inhibition (p < 0.05) of a TNF-α-induced increase in the production of interleukin-6 (IL-6) among 21 tested polyphenolic compounds. p-Coumaric acid, quercetin, and resveratrol demonstrated inhibitions of TNF-α-induced changes in levels of monocyte chemoattractant protein-1 (MCP-1), plasminogen activator inhibitor-1 (PAI-1), and intracellular reactive oxygen species (ROS) in 3T3-L1 adipocytes. Furthermore, p-coumaric acid, quercetin, and resveratrol increased levels (p < 0.05) of secreted adiponectin, superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GPx), and glutathione S-transferase (GST) in TNF-α-treated 3T3-L1 adipocytes. These results indicate that the inhibition of TNF-α-induced changes of adipokines and oxidative stress by some polyphenolic compounds might have further implications in preventing obesity-related pathologies.     

Triacylglycerols, glycerophospholipids, tocopherols, and tocotrienols in berries and seeds of two subspecies (ssp. sinensis and mongolica) of Sea Buckthorn (Hippophaë rhamnoides)

Berries and seeds of two subspecies (ssp. sinensis and mongolica) of sea buckthorn (Hippopha? rhamnoides L.) were compared in terms of triacylglycerols, glycerophospholipids, tocopherols, and tocotrienols. The berries of ssp. mongolica contained less oleic acid (4.6 vs 20.2%, p < 0.001) and more palmitic (33.9 vs 27.4%, p < 0.01) and palmitoleic (32.8 vs 21.9%, p < 0.05) acids in triacylglycerols than those of ssp. sinensis. The proportions of linoleic acid (32.1 vs 22.2%, p < 0.01, in berries; 47.7 vs 42.7%, p < 0.05, in seeds) and palmitic acid (21.1 vs 16.4%, p < 0.001, in berries; 17.0 vs 14.1%, p < 0.05, in seeds) in glycerolphospholipids were higher in ssp. mongolica than in ssp. sinensis, and vice versa with oleic acid (4.3 vs 18.5% in berries, 10.0 vs 22.2% in seeds, p < 0.001). A higher proportion of alpha-linolenic acid was also found in the glycerophospholipids of ssp. sinensis berries (16.2 vs 10.1%, p < 0.001). alpha-, beta-, gamma-, and delta-tocopherols constituted 93-98% of total tocopherols and tocotrienols in seeds, and alpha-tocopherol alone constituted 76-89% in berries. The total contents of tocopherols and tocotrienols varied within the ranges of 84-318 and 56-140 mg kg(-1) in seeds and whole berries, respectively. The seeds of ssp. mongolica were a better source of tocopherols and tocotrienols than those of ssp. sinensis (287 vs 122 mg kg(-1), p < 0.001). The compositional differences between the two subspecies should be considered when the berries are bred and exploited for nutritional purposes.     

Metabolic profiling of strawberry grape ( Vitis × labruscana cv. 'Isabella') components by nuclear magnetic resonance (NMR) and evaluation of their antioxidant and antiproliferative properties

In the assessment of the antioxidant properties of edible plants, the widely consumed Vitis × labruscana cv. 'Isabella', known in Italy as "fragola" (strawberry) grape, was of interest. Phenol and flavonoid contents of the methanolic extracts of peel, pulp, seed, leaf, and stalk components of the plant were determined. The metabolic profile of the extracts was performed by 1D and 2D NMR. Quantitative analysis, obtained in the presence of 0.01% of internal standard trimethylsilyl propionate, evidenced the presence of catechins in both stalk and seed extracts, whereas caffeic acid and quercetin were the main metabolites of the leaf extract. Furthermore, the extracts were tested for their radical scavenging and reducing capacities by measuring their capacity to scavenge DPPH(?) and ABTS(?+) and to reduce Fe(III) and Mo(VI) salts. The antioxidant efficacy of the extracts in cell-free systems and their antiproliferative activity toward HepG2 and A549 cells were also evaluated. Seed and stalk components are able to reduce by 39.6 and 40.6%, respectively, the amount of the metabolically active HepG2 cells after only 24 h of exposure.     

Origin and cytology of a novel cytotype of Allium tuberosum Rottl. ex Spreng. (2n = 48)

Naturally occurring/spontaneously produced polyploids with six/more genomes are rarely found in Alliums. A hexaploid form of Allium tuberosum with 2n = 48 chromosomes has been isolated for the first time amongst the open-pollinated seedlings of a hypotetraploid plant (2n = 4x = 31); latter being the seed-derived product of a normal tetraploid stock (2n = 4x = 32) growing in Jammu University Botanical Garden. Except for the guard cells and pollen grains that are of increased size, this form compared to its progenitor is dwarf, has smaller leaves and bears inflorescences with few flowers. This plant is also different from its progenitor in having nearly one-fourth (27.8 %) of its pollen mother cells (PMCs) with varying chromosome number viz. 27–64, with the remaining cells having somatic or double the somatic number of chromosomes. To assess the nature of hexaploid form, its chromosomes were studied for morphological details, putative grouping and pairing properties during reduction division. Morphological similarity in the chromosomes of the present cytotype and its progenitor, arrangement of 48 chromosomes in eight groups of six chromosomes each and presence of 21.88 % euploid cells with eight hexavalents pointed towards the autopolyploid nature of the present strain. Regarding the origin of this strain, observation made on the meiosis in the two sex mother cells of the progenitor provides some clues. In the later plant, presence of most of embryo-sac mother cells with 62 chromosomes that showed 31:31 segregations and existence of majority of the PMCs with 31 chromosomes exhibiting erratic segregations indicate that the hexaploid strain has probably originated as a result of the fusion of reduced male (n = 17) and unreduced female gamete (2n = 31).     

Photochemical oxidation of polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) in soils — a tool to assess their degradability?

To determine the degradability of PAHs and PCBs for soil remediation or ecotoxicological risk assessment, a simple method is needed. We tested the suitability of photocatalytic oxidation for this purpose. We determined the concentrations of 20 PAHs and 12 PCBs in four mineral topsoil horizons, six organic horizons, and four particle‐size fractions of each of three soils before and after UV irradiation with TiO₂ as a catalyst in suspension. Preliminary experiments showed that in dry soil no photooxidation occurred, but after 48 h of irradiation in suspension the PCB concentrations decreased by up to 40—50 %, while the PAH concentrations did not change significantly. In contrast to this, 95—100 % of PAH and PCB standards spiked on quartz sand were degraded within 8 h, indicating that sorption to organic matter limited degradation of PAHs and PCBs in soil suspensions. There was no difference in the degradation among different individual PAHs and PCBs, respectively, indicating that the degradation did not occur in dissolved state, but in association with soil organic matter. In all samples except one, the degradation of PCBs (10—80 % loss of initial concentrations) was higher than those of the PAHs (0—40 % loss). This suggests that the accessibility of PCBs for OH· radicals generated during irradiation was higher, or the oxidation of PAHs was limited by the properties of the sorbing organic matter. Thus, the tested method was not suitable to predict biodegradability, because it did not reflect the differences in degradability of individual compounds.     

Effects of potassium silicate and nanosilica on quantitative and qualitative characteristics of a commercial strawberry (fragaria × ananassa cv. ‘camarosa’)

Silicon (Si) is considered as a beneficial element to higher plants especially under stress conditions. A factorial experiment, in a completely randomized design with four replications, was used to investigate the effects of two application methods (spraying and soil drenching) and eight levels of Si compounds including 0, 5, 10 and 15 mM of potassium silicate (K₂SiO₃) and 0, 5, 10 and 15 mM of nanosilica (SiO₂). The results indicated that Si application (all or some treatments) decreased transpiration, specific leaf area, petiole length, and promoted the flowering, fruit firmness, leaf/crown number, fresh and dry weight of shoot and root, water use efficiency. They also showed it did not affect the fruit set percentage, yield, chlorophyll index, total soluble solid, leaf area, fresh and dry weight of crown, photosynthesis, stomatal conductance, internal carbon dioxide (CO₂) concentration and mesophyll efficiency of strawberry. The results suggest the beneficial effects of Si on growth and development of strawberry.     

Studies of O,O-dimethyl α-(2,4-dichlorophenoxyacetoxy)ethylphosphonate (HW02) as a new herbicide. 1. Synthesis and herbicidal activity of HW02 and analogues as novel inhibitors of pyruvate dehydrogenase complex

On the basis of the previous work for optimization of O,O-diethyl α-(substituted phenoxyacetoxy)alkylphosphonates, further extensive synthetic modifications were made to the substituents in alkylphosphonate and phenoxy moieties of the title compounds. New O,O-dimethyl α-(substituted phenoxyacetoxy)alkylphosphonates were synthesized as potential inhibitors of pyruvate dehydorogenase complex (PDHc). Their herbicidal activity and efficacy in vitro against PDHc were examined. Some of these compounds exhibited significant herbicidal activity and were demonstrated to be effective inhibitors of PDHc from three different plants. The structure-activity relationships of these compounds including previously reported analogous compounds were studied by examining their herbicidal activities. Both inhibitory potency against PDHc and herbicidal activity of title compounds could be increased greatly by optimizing substituent groups of the title compounds. O,O-Dimethyl α-(2,4-dichlorophenoxyacetoxy)ethylphosphonate (I-5), which acted as a competitive inhibitor of PDHc with much higher inhibitory potency against PDHc from Pisum sativum and Phaseolus radiatus than from Oryza sativa , was found to be the most effective compound against broadleaf weeds and showed potential utility as herbicide.     

High-performance liquid chromatography with photodiode array detection (HPLC-DAD)/HPLC-mass spectrometry (MS) profiling of anthocyanins from Andean Mashua Tubers (Tropaeolum tuberosum Ruíz and Pavón) and their contribution to the overall antioxidant activity

Mashua (Tropaeolum tuberosum Ruíz and Pavón), an Andean tuber with high antioxidant activity, has sparked interest because of its traditional medicinal use. In this study, we evaluated the anthocyanin composition for three purple mashua genotypes and their contribution to the overall antioxidant activity of the tuber. Mashua anthocyanins, total phenolics, and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) antioxidant activity ranged from 45.5 to 131.9 mg of cyanidin 3-glucoside equivalents/100 g fresh weight (FW), 174.9 to 275.5 mg of gallic acid equivalents/100 g of FW, and 16.2 to 45.7 micromol of Trolox equivalents/g of FW, respectively. The high-performance liquid chromatography with photodiode array detection (HPLC-DAD) and HPLC-electrospray ionization tandem mass spectrometry (ESI/MS-MS) profiles revealed the presence of 11 different anthocyanins. The two major pigments (56.4-73.0% total area range at 520 nm) were identified as delphinidin 3-glucoside-5-acetylrhamnoside and delphinidin 3-sophoroside-5-acetylrhamnoside. Other pigments were delphinidin 3-glucoside-5-rhamnoside, delphinidin 3-sophoroside-5-rhamnoside, delphinidin 3-glucoside, cyanidin 3-sophoroside, and cyanidin 3-sophoroside-5-rhamnoside. Cyanidin 3-glucoside and cyanidin 3-rutinoside were only found in two genotypes, while pelargonidin 3-sophoroside and pelargonidin 3-sophoroside-5-rhamnoside were only found in the third one. Anthocyanins from mashua were the major contributors to the total ABTS values for only one of the three genotypes, suggesting that other phenolics present are playing a major role in the antioxidant power of mashua tubers. Results from this study provide important information for the Nutraceutical and Functional Food Market for the use of mashua anthocyanins not only as a source of natural colorants but also as a source of phytonutrients.     

Specific synthesis of 5,5'-dicapsaicin by cell suspension cultures of capsicum annuum var. annuum (chili Jalapeño chigol) and their soluble and NaCl-extracted cell wall protein fractions

HPLC-UV, (1)H NMR, (13)C NMR, and (1)H-(1)H COSY analyses revealed that exogenous capsaicin was specifically converted into 5,5'-dicapsaicin by both cell suspension cultures of Capsicum annuum var. annuum (chili Jalape?o chigol) and their soluble and NaCl-extracted cell wall protein fractions under oxidative conditions. In cell suspension cultures 5,5'-dicapsaicin was found only in biomass of capsaicin-fed cultures. This compound has not been detected before either in fresh fruits or in in vitro cultures of Capsicum. The transformation of capsaicin by different protein fractions revealed that most of the enzymatic activity was located in the NaCl-extracted, or ionic cell wall bound, protein, and that it was strictly dependent on H(2)O(2). These results might in part explain some previously described features of capsaicin production by in vitro cultures of Capsicum. The implications of the results regarding the catabolism of capsaicinoids are discussed.     

Effect of long‐term cultivation on zinc and copper contents in soils from the bahía blanca horticultural belt (Argentina)

Abstract

The objectives of this work were to quantify the total and available copper (Cu) and zinc (Zn) contents in soils which have been used for intensive agricultural production and to investigate the influence of the soil's properties on the absorption and migration of these metals. Total and available Zn and Cu contents in the topsoil and their variation with depth in Mollisols from the Bahía Blanca (Argentina) horticultural belt have been studied. Total contents of Cu and Zn were extracted by means of acid digestion in Teflon bombs placed in a microwave oven. The available Cu and Zn contents were extracted using the DTPA method and the analysis of Cu and Zn was carried out by atomic absorption spectrophotometry. High contents of Cu and Zn were found in the surface layers of all the plots studied and lower contents were noted in plots recently used for horticulture. The lowest contents were found in control farms. The results showed that 70% of the Zn in the surface layers were found in unavailable form, whereas almost 50% of the Cu was in available form. No relationship was noted between each kind of cultivation and Cu content. Most of the Zn in the different plots was adsorbed or complexed by organic ligands and clay. The variation of Cu and Zn content with the depth indicated a greater mobility of Cu. Most of the Zn was detected in the surface soil, whereas no differences were noted in Cu content between surface and subsoil. The results obtained confirmed the anthropic origin of these metals in the cultivated plots. The correlation found between the different forms of Zn and Cu as well as between their total and available contents suggested that these metals have been applied together via agrochemicals and waste.     

Calibration of the Ångström–Prescott coefficients (a,b) under different time scales and their impacts in estimating global solar radiation in the Yellow River basin

The Ångström–Prescott (A–P) equation relating in its current form the incident top-of-atmosphere solar radiation to the solar radiation received at the surface, is one of the most accurate and widely used sunshine-based methods estimating global solar radiation (R_s). The key in its application is the calibration of the locally specific coefficients. Although the coefficients have been extensively studied and calibrated in many places over the world, their relations with time scale are much less investigated. This paper addressed the variation in these coefficients caused by time scale and how this variation results on the accuracy of R_s predictions. This was done using long-term data at 31 sites from the Yellow River basin in China by parallel calibration at three time scales: daily, monthly mean daily and yearly mean daily. We found that the A–P coefficients obtained using monthly data generally had higher a and lower b and larger variations over those using daily data. At yearly time scale, the sunshine–radiation relationship can no longer be described by the linear A–P formula. The difference in coefficients between daily and monthly calibration was rather large accounting for 71% of the differences in a and 61% in b that in turn were greater than 0.03, corresponding to 81% for a and 49% for b being greater than 10%. Time scale had a larger effect on a than on b, and it caused a maximum variation of 82% in a and 43% in b in the basin, equaling half of the variation caused by geographical location. However, the large effect of time scales on a and b produced no significant impact on the estimation accuracy of R_s because of the conservative response of the sum a + b to time scale. In this sense, the coefficients calibrated at daily scale are interchangeable with those calibrated at monthly scale, indicating the high flexibility of the A–P formula. Nevertheless, calibration made at daily scale has two important advantages over monthly scale in that it requires fewer years’ data to obtain stabilized coefficients, and that it is easily predicted more accurately with common site information. Our findings have two implications. Firstly, they provide an additional guidance on the explanation of the large variability of the coefficients found in the literature for the same geographical location. Secondly, they facilitate the choice of the coefficients in practical applications by proving their interchangeability in the estimation of R_s.     

Genetics of Wild and Managed Populations of <Emphasis Type="Italic">Leucaena esculenta</Emphasis> Subsp. <Emphasis Type="Italic">esculenta</Emphasis> (Fabaceae; Mimosoideae) in La Montaña of Guerrero,Mexico

Genetic consequences of silvicultural management of Leucaena esculenta subsp. esculenta were analyzed from eight allozyme loci in half-sib families of one wild and one managed in situ (selectively cleared) population from La Montaña de Guerrero region, Central Mexico. A reference sample (including wild, feral and cultivated individual plants) from the states of Morelos, Puebla and Guerrero, Mexico was also analyzed. Genetic variation, population structure and mating system were analyzed. All loci showed high variation (75–87.5% polymorphic loci at 95% level; 2.4–2.8 mean number of alleles per locus). All progenies showed heterozygous deficiency, but both wild and managed parental inbreeding coefficients were negative, suggesting heterosis. Progenies of managed populations differed from those of the wild and reference samples (Nei’s unbiased identities 0.874–0.934). Biparental inbreeding is suggested by Wright’s-statistics ( f = 0.313), and by outcrossing rate estimates: t_m = 0.644 (SE 0.094), and 0.645 (SE 0.193); t_s = 0.576 (SE 0.189), and 0.523 (SE 0.182), for managed and wild samples respectively. Population differentiation is significant (Θ = 0.210). The species is self-incompatible and deviations from the mixed mating model were found. Indirect estimates of products of effective population size (N_e) by the proportion of migrants (N_m) were moderate, as were the N _evalues. Variation due to ecotypic differentiation (related to altitude), prolonged artificial selection, and introduction from other areas is supported. A model of domestication of seed-propagated trees is suggested, based on extensive and in situ selection of locally adapted populations, and their diffusion to other areas.