Anatomy and cell wall polysaccharides of almond (Prunus dulcis D. A. Webb) seeds

The anatomy of Prunus dulcis was analyzed by applying several differential staining techniques and light microscopy. Prunus dulcis seed has a thin and structurally complex seed coat, with lignified cellulosic tissue. The embryo has two voluminous cotyledons. Cotyledon cells have a high number of protein and lipid bodies, some of which have phytin. The provascular tissue, located in the cotyledons, is oriented in small bundles perpendicular to the transverse embryonic axis. Prunus dulcis cell wall material is very rich in arabinose (45 mol %). Glucose (23%), uronic acids (12%), and xylose (12%) are also major sugar components. The polymers obtained from the imidazole and Na(2)CO(3) extracts contain mainly pectic substances rich in arabinose, but the sugar content of these extracts was very low. The majority of the pectic substances (also rich in arabinose) was recovered with the KOH extracts. These extracts, with high sugar content, yielded also xyloglucans and acidic xylans. The 4 M KOH + H(3)BO(3) extracts yielded polysaccharides rich in uronic acids and xylose and very rich in arabinose, accounting for 27% of the cell wall material.     

Almond (Prunus dulcis (Mill.) D.A. Webb) skins as a potential source of bioactive polyphenols

An exhaustive study of the phenolic composition of almond ( Prunus dulcis (Mill.) D.A. Webb) skins was carried out in order to evaluate their potential application as a functional food ingredient. Using the HPLC-DAD/ESI-MS technique, a total of 33 compounds corresponding to flavanols, flavonols, dihydroflavonols and flavanones, and other nonflavonoid compounds were identified. Peaks corresponding to another 23 structure-related compounds were also detected. MALDI-TOF MS was applied to characterize almond skin proanthocyanidins, revealing the existence of a series of A- and B-type procyanidins and propelargonidins up to heptamers, and A- and B-type prodelphinidins up to hexamers. Flavanols and flavonol glycosides were the most abundant phenolic compounds in almond skins, representing up to 38-57% and 14-35% of the total quantified phenolics, respectively. Due to their antioxidant properties, measured as oxygen-radical absorbance capacity (ORAC) at 0.398-0.500 mmol Trolox/g, almond skins can be considered as a value-added byproduct for elaborating dietary antioxidant ingredients.     

Antiproliferative terpenoids from almond hulls (Prunus dulcis): identification and structure-activity relationships

Bioassay-guided fractionation of the EtOAc crude extract from Sicilian almond hulls, a waste material from Prunus dulcis crop, allowed identification of 10 constituents, isolated as pure compounds (1-5, 7, and 10) or unseparable mixtures (5 + 6 and 8 + 9). All compounds were subjected to spectroscopic analysis and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide bioassay on MCF-7 human breast cancer cells. In addition to the main components oleanolic (1), ursolic (2), and betulinic (3) acids, the 2-hydroxy analogues alphitolic (4), corosolic (5), and maslinic (6) acids, as well as the related aldehydes, namely, betulinic (7), oleanolic (8), and ursolic (9), were identified. From a more polar fraction, the beta-sitosterol 3-O-glucoside (10) was also identified. A sample of commercially available betulin (11) was also included in bioassays as further support to a structure-activity relationship study. Betulinic acid showed antiproliferative activity toward MCF-7 cells (GI50 = 0.27 microM), higher than the anticancer drug 5-fluorouracil.     

Fostering sustainable water use in almond (Prunus dulcis Mill.) orchards in a semiarid Mediterranean environment

This work examines the long-term effects of deficit-irrigation (DI) practices in almond crop (Prunus dulcis Mill.) in agronomical and physiological terms. The trial was conducted during four-year monitoring period (2014–2017), in an experimental orchard (SW Spain), subjected to three irrigation regimes; i) a full-irrigation treatment (F_I), which received 100% of crop evapotranspiration (ET_C); ii) a regulated-deficit irrigation (RDI₅₀), which received 50% of ET_C during the kernel-filling period; and iii) a low-frequency deficit irrigation (LFDI), that was subjected to continuous periods of irrigation-restriction defined in terms of threshold values of leaf-water potential (Ψ_leaf) during the kernel-filling period. During the water stress period, there were monitored Ψ_leaf, stomatal conductance (g_s) and canopy temperature (T_C). Significant improvements in terms of water-use efficiency were found, as no differences in terms of yield between F_I and LFDI were found, leading to the conclusion that significant water savings (between 27 and 40%) can be achieved without compromising the yield. Moreover, threshold values of Ψ_leaf and thermal indicators were defined which will allow establishing future irrigation scheduling without compromising almond yield, especially when DI strategies are being applied.     

New prenylated benzoic acid and other constituents from almond hulls (Prunus amygdalus Batsch).

One new prenylated benzoic acid derivative, 3-prenyl-4-O-beta-D-glucopyranosyloxy-4-hydroxylbenzoic acid, and three known constituents, catechin, protocatechuic acid, and ursolic acid, have been isolated from the hulls of almond (Prunus amygdalus). Complete assignments of the proton and carbon chemical shifts for the new prenylated benzoic acid derivative were accomplished on the basis of high-resolution 1D and 2D nuclear magnetic resonance data. All of these compounds except ursolic acid are being reported from almond hulls (P. amygdalus) for the first time.     

Production and characterization of rabbit polyclonal antibodies to almond (Prunus dulcis L.) major storage protein.

Rabbits were immunized with purified almond major protein (AMP), the primary storage protein in almonds. Rabbit anti-AMP polyclonal antibodies (PA) could detect AMP when as little as 1-10 ng/mL were used to coat microtiter plates in a noncompetitive enzyme linked-immunosorbent assay (ELISA). Competitive inhibition ELISA assays detected the AMP down to 300 ng/mL. PA recognized the AMP in protein extracts from all U.S. major marketing cultivars of almonds (Mission, Neplus, Peerless, Carmel, and Nonpareil) with specific reactivity of 52.6-75% as compared to that of the AMP alone. Immunoreactivity of protein extracts prepared from commercial samples of blanched almonds, roasted almonds, and almond paste was respectively reduced by 50.0%, 56.6%, and 68.4% (noncompetitive ELISA) when compared to the immunoreactivity of the AMP. Moist heat (121 degrees C, 15 min) pretreatment of the AMP reduced the PA reactivity by 87% (noncompetitive ELISA). Exposing AMP to pH extremes (12.5 and 1.5-2.5) caused a 53% and 57% reduction in PA reactivity, respectively (noncompetitive ELISA). PA showed some cross-reactivity with the cashew major globulin, and to a lesser extent, the Tepary and Great Northern bean major storage protein (7S or phaseolin). The presence of almonds in a commercial food was detected using PA in a competitive ELISA.     

Biochemical characterization of amandin,the major storage protein in almond (Prunus dulcis L.)

The almond major storage protein, amandin, was prepared by column chromatography (amandin-1), cryoprecipitation (amandin-2), and isoelectric precipitation (amandin-3) methods. Amandin is a legumin type protein characterized by a sedimentation value of 14S. Amandin is composed of two major types of polypeptides with estimated molecular weights of 42-46 and 20-22 kDa linked via disulfide bonds. Several additional minor polypeptides were also present in amandin. Amandin is a storage protein with an estimated molecular weight of 427,300 +/- 47,600 Da (n = 7) and a Stokes radius of 65.88 +/- 3.21 A (n = 7). Amandin is not a glycoprotein. Amandin-1, amandin-2, and amandin-3 are antigenically related and have similar biochemical properties. Amandin-3 is more negatively charged than either amandin-1 or amandin-2. Methionine is the first essential limiting amino acid in amandin followed by lysine and threonine.     

Antioxidant activity of prune (Prunus domestica L.) constituents and a new synergist

Ethanol extract of prune was separated into hexane-soluble and H(2)O-soluble fractions, and the H(2)O-soluble fraction was further separated into a methanol (MeOH) eluate and an H(2)O eluate by Diaion HP-20 column chromatography. The MeOH eluate exhibited the strongest antioxidant activity among the separated fractions evaluated by oxygen radical absorbance capacity (ORAC). Further purification of the MeOH eluate led to isolation of a novel compound, 4-amino-4-carboxychroman-2-one, together with four known compounds (p-coumaric acid, vanillic acid beta-glucoside, protocatechuic acid, and caffeic acid), structures of which were identified by NMR and MS analyses. The ORAC values of these isolated compounds showed 0.15-1.43 units (micromol of Trolox equiv)/micromol, and the new compound showed a remarkable synergistic effect on caffeoylquinic acid isomers. The antioxidant activity of the MeOH eluate was highly dependent on the major prune components, caffeoylquinic acid isomers, with a contribution from the new synergist.     

Effects of roasting,blanching, autoclaving,and microwave heating on antigenicity of almond (Prunus dulcis L.) proteins

Whole, unprocessed Nonpareil almonds were subjected to a variety of heat processing methods that included roasting (280, 300, and 320 degrees F for 20 and 30 min each; and 335 and 350 degrees F for 8, 10, and 12 min each), autoclaving (121 degrees C, 15 psi, for 5, 10, 15, 20, 25, and 30 min), blanching (100 degrees C for 1, 2, 3, 4, 5, and 10 min), and microwave heating (1, 2, and 3 min). Proteins were extracted from defatted almond flour in borate saline buffer, and immunoreactivity of the soluble proteins (normalized to 1 mg protein/mL for all samples) was determined using enzyme linked immunosorbent assay (ELISA). Antigenic stability of the almond major protein (amandin) in the heat-processed samples was determined by competitive inhibition ELISA using rabbit polyclonal antibodies raised against amandin. Processed samples were also assessed for heat stability of total antigenic proteins by sandwich ELISA using goat and rabbit polyclonal antibodies raised against unprocessed Nonpareil almond total protein extract. ELISA assays and Western blotting experiments that used both rabbit polyclonal antibodies and human IgE from pooled sera indicated antigenic stability of almond proteins when compared with that of the unprocessed counterpart.     

Sphingolipid and other constituents from almond nuts (Prunus amygdalus Batsch)

One sphingolipid, 1-O-beta-D-glucopyranosyl-(2S,3R,4E,8Z)-2-[(2R)-2-hydroxyhexadecanoylamino]-4,8-octadecadiene-1,3-diol, and four other constituents, beta-sitosterol, daucosterol, uridine, and adenosine, have been isolated from the nuts of almond (Prunus amygdalus). Complete assignments of the proton and carbon chemical shifts for the sphingolipid were accomplished on the basis of high-resolution 1D and 2D NMR data. All of these compounds are being reported from almond nuts (P. amygdalus)for the first time.     

Population structure and phylogenetic relationship of Peach [Prunus persica (L.) Batsch] and Nectarine [Prunus persica var. nucipersica (L.) C.K. Schneid.] based on retrotransposon markers

For effective varietal improvement of horticultural crops peach (Prunus persica) and nectarine (Prunus persica var. nucipersica), information about their population structure and genetic relatedness plays an important role. In this study we used retrotransposon-based markers (iPBS) to estimate the genetic diversity and population structure of 48 peach and nectarine genotypes from various distinct geographical regions of Punjab and Khyber Pakhtunkhwa, Pakistan. A total of 461 alleles were identified from PCR amplicons derived from nine iPBS primer pairs with an average of 8.5 alleles/locus. Among all four groups the genotypes collected from Swat and Hunza had the highest and the lowest expected heterozygosity, unbiased expected heterozygosity and Shannon’s information index, respectively. We constructed a Neighbour-Joining dendrogram and performed principal coordinate analysis based on the distance matrices, and both forms of analysis grouped the 48 genotypes into two distinct clusters. The STRUCTURE software distributed the forty-eight genotypes into two main populations (k?=?2) indicating a low diversity between genotypes collected from Chakwal, Swat, Mansehra and Hunza.

     

Molecular Characterization of Local Spanish Peach [Prunus persica (L.) Batsch] Germplasm

A collection of 85 local Spanish peach cultivars has been studied with SSR markers. To carry out this work 42 SSRs previously developed in peach were screened to select the microsatellite loci that presented high polymorphism and that were easier to score in high resolution agarose gels. Ten SSRs were selected to identify the peach cultivars and to estimate the genetic similarity among them. The selected microsatellites resulted in the amplification of 35 fragments in the 85 genotypes studied, differentiating 46 genotypic profiles, from which 31 correspond to unique cultivars. The rest of the profiles correspond to groups of two to five cultivars and one group of 17 undistinguishable cultivars. The genetic similarity estimated from the molecular data clearly separated the flat and most of the white-fleshed peaches from the rest of genotypes that are mostly yellow non-melting local peaches. The results obtained in this work will help to improve the conservation and management of local Spanish peach germplasm allowing the establishment of core collections that represent most of the diversity conserved.     

Purification, crystallization and preliminary X-ray characterization of prunin-1, a major component of the almond (Prunus dulcis) allergen amandin

The 11S globulins from plant seeds account for a number of major food allergens. Because of the interest in the structural basis underlying the allergenicity of food allergens, we sought to crystallize the main 11S seed storage protein from almond ( Prunus dulcis). Prunin-1 (Pru1) was purified from defatted almond flour by water extraction, cryoprecipitation, followed by sequential anion exchange, hydrophobic interaction, and size exclusion chromatography. Single crystals of Pru1 were obtained in a screening with a crystal screen kit, using the hanging-drop vapor diffusion method. Diffraction quality crystals were grown after optimization. The Pru1 crystals diffracted to at least 3.0 A and belong to the tetragonal space group P4(1)22, with unit cell parameters of a = b = 150.912 A, c = 165.248 A. Self-rotation functions and molecular replacement calculations showed that there are three molecules in the asymmetry unit with water content of 51.41%. The three Pru1 protomers are related by a noncrystallographic 3-fold axis and they form a doughnut-shaped trimer. Two prunin trimers form a homohexamer. Elucidation of prunin structure will allow further characterization of the allergenic features of the 11S protein allergens at the molecular level.     

Antioxidant constituents in the fruits of Luffa cylindrica (L.) Roem

Hydrophilic antioxidant constituents in the fruits of the vegetable Luffa cylindrica (L.) Roem (sponge gourds) were separated by an antioxidant-guided assay to yield eight compounds: p-coumaric acid (1), 1-O-feruloyl-beta-D-glucose (2), 1-O-p-coumaroyl-beta-D-glucose (3), 1-O-caffeoyl-beta-D-glucose (4), 1-O-(4-hydroxybenzoyl)glucose (5), diosmetin-7-O-beta-D-glucuronide methyl ester (6), apigenin-7-O-beta-D-glucuronide methyl ester (7), and luteolin-7-O-beta-D-glucuronide methyl ester (8). The eight compounds were isolated by high-speed countercurrent chromatography and identified by electrospray ionization-mass spectrometry and NMR analysis, and the antioxidant activity was evaluated by the radical scavenging effect on the 1,1-diphenyl-2-picrylhydrazyl radical. High-performance liquid chromatography analysis showed that a total amount of the eight compounds in the dried gourds without skin was about 1%. The results demonstrate that the consumption of sponge gourds can supply some antioxidant constituents to human body.     

Isolation and characterization of endogenous inhibitors of nitrate reductase of peach [Prunus persica (l.) Batsch] and their distribution in the genus Prunus 1

Endogenous inhibitors of nitrate reductase (NR) of peach [Prunus persica (L.) Batsch] seedlings were identified as mandelonitrile and cyanide. The compounds gave greater than 90% inhibition at 0.019 μM and were noncompetitive with respect to nitrate when preincubated with the enzyme in the absence of nitrate. Competitive inhibition in respect to nitrate occurred when nitrate was present prior to the addition of enzyme. Concentration and distribution of NR inhibitors were: leaf > stem > roots as determined by two independent methods. Nitrate reductase inhibitor was detected in 12 diverse species of Prunus and was tentatively identified as cyanide by high‐performance liquid chromatography (HPLC) and biological activity. The degradation products of the cyanogenic‐glucoside amygalin were tested to determine NR inhibition. Amygdalin, prunasin and benzaldehyde were not inhibitors of NR, whereas mandelonitrile and cyanide were. Prunasin was determined to be the precursor of the NR inhibitors in peach leaf, stem, and root tissue.     

桃果实成熟期主效基因的SSR标记定位

桃作为果树中的模式植物,许多重要的质量性状基因已被定位,呈数量性状遗传的果实成熟期是桃的重要经济性状。本研究以大久保桃(Prunus persica(L.)Okubo)自交后代为群体,通过群体分离分析法(bulk segregation analysis,BSA)法对果实成熟期主效基因进行简单重复序列(SSR)标记,在165对SSR引物中筛选出3对与桃果实成熟期相关,分别为:显性标记UDP96-003和共显性标记BPPCT015、UDP97-402。通过对随机群体中的电泳结果进行FsLinkageMap2.0软件分析和单标记方差分析,得出3个分子标记在一个连锁群上,相对顺序依次为UDP96-003、UDP97-402和BPPCT015,且两两间遗传距离分别为21和13.2cM,且均与果实成熟期主效基因连锁。用FsQtlMap软件对控制桃果实成熟期基因进行区间定位,得出在BPPCT015和UDP97-402之间存在着一个控制桃果实成熟期的主效数量性状基因座(QTL),其LOD值为13.35,效应值达0.623,对果实成熟期的表型平均遗传值分别为qq:84.59d、Qq:100.08d和QQ:115.25d,即该位点单基因效应值约15d。该研究结果对前人的推论提供了实验证据,并将该基因锚定在两个SSR标记之间。     

Zur Gliederung vonPetroselinum crispum (Mill.) Nym.

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美国山核桃总多酚与总黄酮含量及抗氧化活性

为研究不同种质美国山核桃中总多酚与总黄酮含量及抗氧化活性的差异,对29个美国山核桃种质脱脂种仁的总多酚、总黄酮含量及抗氧化活性进行测定和比较。结果表明,29个美国山核桃种质脱脂种仁的总多酚、总黄酮含量及抗氧化活性存在显著差异,其中ZL86号种质总多酚含量最高,为47.96mg GAE·g~(-1),且抗氧化活性最强;ZL65号种质总黄酮含量最高,为24.01 mg RE·g~(-1);总多酚、总黄酮含量均与抗氧化活性呈极显著正相关。聚类分析将29个种质分为2类:Ⅰ类包含18个种质,其总多酚含量、总黄酮含量和抗氧化活性均较低;Ⅱ类包含11个种质,其总多酚含量、总黄酮含量和抗氧化活性均较高。本研究为美国山核桃品种的选育提供了一定的理论依据。     

Mutanten der WildtomateLycopersicon pimpinellifolium (Jusl.) Mill. I

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