Evaluación de la capacidad antioxidante y compuestos bioactivos de tumbo (Passiflora mollissima) y cerezo (Prunus serotina)

Objetivo: Determinar el contenido de compuestos bioactivos y la capacidad antioxidante de los frutos tumbo (Passiflora mollissima) y cerezo (Prunus serotina). Materiales y métodos: Estudio analítico, experimental, longitudinal y prospectivo. Los frutos cerezo y tumbo se recolectaron en las regiones de Cusco, Moquegua y Arequipa. La técnica de Folin-Ciocalteu fue empleada para determinar el contenido de fenoles, y el cloruro de aluminio se utilizó para calcular los flavonoides. La actividad antioxidante se evaluó mediante las técnicas Ferric Reducing Ability of Plasma (FRAP), 2,2-difenil-picril-hidrazil (DPPH) y sustancias reactivas al ácido tiobarbitúrico (TBARS). Para estudiar el efecto hepatoprotector de las frutas, utilizamos ratas albinas que se clasificaron en un grupo control negativo, un grupo control positivo y cuatro grupos experimentales. Resultados: La mayor concentración de fenoles totales y flavonoides se encontró en el tumbo de la región Cusco (Quechua) (fenoles totales: 584,94 ± 134,62 mg EAG / 100 g y flavonoides :445,62 ± 7,94 mg EQ / 100 g). Para el radical DPPH, el valor IC50 del tumbo de la región Arequipa (Yunga) fue 0,41 ± 0,01 mg / mL. El tumbo de la región Cusco (Quechua) mostró el valor FRAP más alto (8,38 ± 0,32 mmol Fe2 + / 100 g). El cerezo de la región de Arequipa (Yunga) presentó la mayor concentración de fenoles totales (181,81 ± 34,1 mg EAG / 100 g) y flavonoides (205,18 ± 77,8 mg EQ / 100 g). El cerezo de Arequipa (Yunga) mostró una actividad antioxidante significativa al DPPH (2,1 ± 0,01 mg / mL), mientras que la capacidad antioxidante del cerezo de la región Cusco (Quechua), evaluada con la técnica FRAP, alcanzó un valor de 1,59 ± 0,2 mmol Fe2+/100 g. Las diferencias observadas fueron estadísticamente significativas. El tumbo mostró un mejor efecto hepatoprotector que el cerezo. Conclusiones: El tumbo de la región Cusco (Quechua) es una fuente importante de compuestos antioxidantes y muestra una elevada capacidad antioxidante (FRAP), mientras que el cerezo de la región Arequipa (Yunga) tiene un alto contenido de compuestos antioxidantes y una mayor capacidad antioxidante (DPPH).

Objective: To determine the content of bioactive compounds and antioxidant capacity of the banana passionfruit (Passiflora mollissima) and black cherry (Prunus serotina). Materials and methods: An analytical, experimental, longitudinal and prospective study. The black cherries and banana passionfruits were collected in the Cusco, Moquegua and Arequipa regions. The content of phenols and flavonoids were determined using the Folin-Ciocalteu reagent and aluminum chloride method, respectively. The antioxidant activity was evaluated using the ferric reducing ability of plasma (FRAP), 2,2-diphenyl-1-picryl-hydrazyl (DPPH) and thiobarbituric acid reactive substance (TBARS) techniques. Albino rats classified into a negative control group, a positive control group and four experimental groups were used to study the hepatoprotective effect of the fruits. Results: Banana passionfruits from the Cusco region (Quechua) showed the highest concentration of total phenols (584.94 ± 134.62 mg GAE/100 g) and flavonoids (445.62 ± 7.94 mg QE/100 g). Concerning the DPPH radical, the IC 50 value of banana passionfruits from the Arequipa region (Yunga) was found to be 0.41 ± 0.01 mg/mL. Banana passionfruits from the Cusco region (Quechua) showed the highest FRAP value (8.38 ± 0.32 mmol Fe2+/100 g). Black cherries from the Arequipa region (Yunga) had the highest concentration of total phenols (181.81 ± 34.1 mg GAE/100 g) and flavonoids (205.18 ± 77.8 mg QE/100 g). They also showed a significant antioxidant activity regarding the DPPH (2.1 ± 0.01 mg/mL), while the antioxidant capacity of black cherries from the Cusco region (Quechua), which was evaluated with the FRAP method, achieved a value of 1.59 ± 0.2 mmol Fe2+/100 g. The observed differences were statistically significant. Banana passionfruits showed a better hepatoprotective effect than black cherries. Conclusions: Banana passionfruits from the Cusco region (Quechua) are an important source of antioxidant compounds and show a high antioxidant capacity (FRAP), while black cherries from the Arequipa region (Yunga) have a high content of antioxidant compounds and a higher antioxidant capacity (DPPH).

Valorization of Almond (Prunus serotina) by Obtaining Bioactive Compounds.

The Capulin almond is a seed of the Prunus serotina (var. capuli) that belongs to the Rosaceae family. In this study, the valorization of the Capulin almond was performed by extracting antioxidants contained in the shell, paste, and oil (extracted by manual cold pressing process) of Prunus serotina treated with methanol, ethanol, acetone, and acidified water (pH 4) in a ratio of 1:5 (w/v). Total phenols were performed using the Folin-Ciocalteu method and expressed as gallic acid equivalents (GAE), antioxidant activity was determined by ABTS and DPPH methods and expressed as Trolox equivalents (TE). Finally, the total flavonoids were determined using a catechin calibration curve and reported as catechin equivalents (CE). The highest extraction of total phenols in shell was obtained with methanol (1.65 mg GAE/g sample) and the lowest using acidified water (0.97 mg GAE/g sample). However, extraction with acidified water favored this process in the paste (1.42 mg GAE/g sample), while the use of solvents did not influence it significantly (0.72 to 0.79 mg GAE/g sample). Regarding the total flavonoids, the values for the shell, paste, and oil were of 0.37, 0.78, and 0.34 mg CE/g sample, respectively, while that corresponding to the antioxidant activity evaluated with ABTS and DPPH were of 1527.78, 1229.17, 18894.44 µM TE/g, and, 568.45, 562.5 and 4369.05 mM TE/g sample, respectively. Finally, our results suggest that by-products such as the shell, paste, and oil obtained from Prunus serotina (var. capuli) represent a potential alternative for the recovery of bioactive compounds with antioxidant activity such as phenolic compounds and flavonoids.

Gametophytic self-incompatibility in Andean capuli (Prunus serotina subsp. capuli): allelic diversity at the S-RNase locus influences normal pollen-tube formation during fertilization.

Capuli (Prunus serotina subsp. capuli) is a tree species that is widely distributed in the northern Andes. In Prunus, fruit set and productivity appears to be limited by gametophytic self-incompatibility (GSI) which is controlled by the S-Locus. For the first time, this research reveals the molecular structure of the capuli S-RNase (a proxy for S-Locus diversity) and documents how S-Locus diversity influences GSI in the species. To this end, the capuli S-RNase gene was amplified and sequenced in order to design a CAPS (Cleaved Amplified Polymorphic Sequence) marker system that could unequivocally detect S-alleles by targeting the highly polymorphic C2-C3 S-RNase intra-genic region. The devised system proved highly effective. When used to assess S-Locus diversity in 15 P. serotina accessions, it could identify 18 S-alleles; 7 more than when using standard methodologies for the identification of S-alleles in Prunus species. CAPS marker information was subsequently used to formulate experimental crosses between compatible and incompatible individuals (as defined by their S-allelic identity). Crosses between heterozygote individuals with contrasting S-alleles resulted in normal pollen tube formation and growth. In crosses between individuals with exactly similar S-allele identities, pollen tubes often showed morphological alterations and arrested development, but for some (suspected) incompatible crosses, pollen tubes could reach the ovary. The latter indicates the possibility of a genotype-specific breakdown of GSI in the species. Overall, this supports the notion that S-Locus diversity influences the reproductive patterns of Andean capuli and that it should be considered in the design of orchards and the production of basic propagation materials.

Phytopharmacological Possibilities of Bird Cherry Prunus padus L. and Prunus serotina L. Species and Their Bioactive Phytochemicals.

Wild cherry is a plant observed in the form of trees or shrubs. This species comprises about twenty kinds of plants and the most popular are two, Prunus padus L. and Prunus serotina L., whose properties and content of phytochemical compounds are subject to studies. Wild cherry contains many active compounds, including tocopherols, vitamins, polyphenols and terpenes, which can have beneficial effects on health. On the other hand, wild cherry contains cyanogenic glycosides. Nevertheless, current research results indicate pro-health properties associated with both P. serotina and P. padus. The aim of this study was to collect and present the current state of knowledge about wild cherry and to review available in vitro and in vivo studies concerning its antioxidant, anti-inflammatory, antibacterial and antidiabetic activity. Moreover, the current work presents and characterizes phytochemical content in the leaves, bark and fruits of P. padus and P. serotina and compiles data that indicate their health-promoting and functional properties and possibilities of using them to improve health. We find that the anatomical parts of P. padus and P. serotina can be a valuable raw material used in the food, pharmaceutical and cosmetic industries as a source of bioactive compounds with multi-directional action.

Decrease in Bat Diversity Points towards a Potential Threshold Density for Black Cherry Management: A Case Study from Germany.

In times of land use changes towards more close-to-nature forestry, the application of bioindicators becomes an interesting tool for effective land-use management schemes. Forest managers are increasingly confronted by alien tree species. Therefore, this case study aimed to investigate the influence of the invasive black cherry (Prunus serotina) on bats (Chiroptera: Verpertilionidae) in pine (Pinus sylvestris) forest ecosystems, in order to identify the potential of bats as bioindicators for a black cherry invasion. In three pre-classified succession stages of the black cherry, the diversity and relative abundance of bats were bioacoustically monitored for a period of 60 nights. From the bat call recordings made during the study period, eight bat species could be identified to species level. Within the succession stages of pine monoculture and light black cherry forest, a comparable bat diversity of eight bat species and three sonotypes with a similar relative abundance were observed. In dense black cherry forest, only four species and one sonotype were detected. Compared to the pine monoculture and light black cherry forest, the overall abundance of the bat community was significantly lower in the dense black cherry forest. Upon evaluation, those bat species associated with the edge and narrow space forager guilds were found to have a high sensitivity to a dense black cherry understory within naturally monocultural pine stands. Their activity patterns indicate that the transition from light to dense black cherry understory can be considered as a potential threshold value for a close-to-nature black cherry understory density in high canopy pine forest stands.

Nitrogen deposition and climate change effects on tree species composition and ecosystem services for a forest cohort.

The composition of forests in the northeastern United States and the ecosystem services they provide to future generations will depend on several factors. In this paper, we isolate the effects of two environmental drivers, nitrogen (N) deposition and climate (temperature and precipitation) change, through an analysis of a single cohort of 24 dominant tree species. We assembled a tree database using data from U.S. Forest Service Forest Inventory and Analysis monitoring plots. Applying observed species-specific growth and survival responses, we simulated how forest stands in a 19-state study area would change from 2005 to 2100 under 12 different future N deposition-climate scenarios. We then estimated implications for three selected forest ecosystem services: merchantable timber, aboveground carbon sequestration, and tree diversity. Total tree biomass (for 24 species combined) was positively associated with both increased N deposition and temperatures; however, due to differences in the direction and magnitude of species-specific responses, forest composition varied across scenarios. For example, red maple (Acer rubrum) trees gained biomass under scenarios with more N deposition and more climate change, whereas biomass of yellow birch (Betula alleghaniensis) and red pine (Pinus resinosa) was negatively affected. Projections for ecosystem services also varied across scenarios. Carbon sequestration, which is positively associated with biomass accumulation, increased with N deposition and increasing climate change. Total timber values also increased with overall biomass; however, scenarios with increasing climate change tended to favor species with lower merchantable value, whereas more N deposition favored species with higher merchantable value. Tree species diversity was projected to decrease with greater changes in climate (warmer temperatures), especially in the northwestern, central, and southeastern portions of the study area. In contrast, the effects of N deposition on diversity varied greatly in magnitude and direction across the study area. This study highlights species-specific and regional effects of N deposition and climate change in northeastern U.S. forests, which can inform management decision for air quality and forests in the region, as well as climate policy. It also provides a foundation for future studies that may incorporate other important factors such as multiple cohorts, sulfur deposition, insects, and diseases.

A rangewide herbarium-derived dataset indicates high levels of gene flow in black cherry (Prunus serotina).

Isolation by Distance (IBD) is a genetic pattern in which populations geographically closer to one another are more genetically similar to each other than populations which are farther apart. Black cherry (Prunus serotina Ehrh.) (Rosaceae) is a forest tree species widespread in eastern North America, and found sporadically in the southwestern United States, Mexico, and Guatemala. IBD has been studied in relatively few North American plant taxa, and no study has rigorously sampled across the range of such a widespread species. In this study, IBD and overall genetic structure were assessed in eastern black cherry (P. serotina Ehrh. var. serotina), the widespread variety of eastern North America. Eastern North America. Prunus serotina Ehrh. var. serotina (Rosaceae). Dense sampling across the entire range of eastern black cherry was made possible by genotyping 15 microsatellite loci in 439 herbarium samples from all portions of the range. Mantel tests and STRUCTURE analyses were performed to evaluate the hypothesis of IBD and genetic structure. Mantel tests demonstrated significant but weak IBD, while STRUCTURE analyses revealed no clear geographic pattern of genetic groups. The modest geographic/genetic structure across the eastern black cherry range suggests widespread gene flow in this taxon. This is consistent with P. serotina's status as a disturbance-associated species. Further studies should similarly evaluate IBD in species characteristic of low-disturbance forests.

The Effect of Size of Black Cherry Stumps on the Composition of Fungal Communities Colonising Stumps.

We investigated fungal communities colonising black cherry stumps. We tested the hypothesis that black cherry stumps of greater diameter should be characterised by more diverse fungal communities than stumps of smaller diameter. The material for analyses came from Podanin Forest District. DNA was extracted using a Plant Genomic DNA purification kit. The results were subjected to bioinformatic analysis and statistical analysis. The OTU sequences were compared using the BLAST algorithm with reference sequences from the UNITE database. In total, 8192 raw sequences were obtained from samples of black cherry stumps applying the Illumina sequencing technique. The results of the statistical analysis indicate a trend towards increased diversity in bigger black cherry stumps. The dominant share of fungi associated with wood decomposition indicates the progressing process of decomposition in stumps. Identification of the role and functions of the individual components of fungal communities colonising stumps may provide insight into the overall ecology of these organisms and provide a basis for improved plant protection, with a view to limiting the occurrence of black cherries in the future in undesirable locations outside their natural range.

Particulate matter on two Prunus spp. decreases survival and performance of the folivorous beetle Gonioctena quinquepunctata.

Woody plants growing along streets and construction sites play an important role in removing harmful particulate matter (PM). Researchers rarely consider the impact of different types and size fractions of PM deposited on the leaves on insect folivores. We determined differences in the accumulation of cement and roadside PM on the leaves of two Prunus species (P. padus and P. serotina) with different leaf surface structures. We also determined the effect of PM on the beetle Gonioctena quinquepunctata, the main pest of these plants. Saplings were artificially dusted in greenhouses and leaves were utilised for larval and adult insect stages feeding in laboratory conditions. Road PM accumulated in greater amounts than did cement PM, regardless of plant species. For both PM sources, P. padus accumulated twofold more than did P. serotina. Insect survival was negatively affected by PM pollution; however, neither Prunus species nor PM source variant significantly affected masses of larvae and pupae, duration of larval and pupal development or relative growth rates. The experiment showed strong negative influences of PM were noted only for adult insects, due to the grazing period being longer than that in larvae. The mass of adult insects and the efficiency of conversion of ingested food (ECI) were lower for insects exposed to PM than those for control insects. Insects compensated for lower ECI by eating a greater total amount of food (TFE). Adult insects gained significantly higher mass when fed with P. serotina than with P. padus. The effect of PM on analysed plant metabolites was insignificant. Only Prunus sp. and date of collection affected the level of condensed tannins and total phenols. Our results indicate that, when investigating the effect of the host plant on folivore performance, the accumulation of PM, as well as its type and quantity, should be taken into account.

In vitro and in vivo cysticidal activity of extracts and isolated flavanone from the bark of Prunus serotina: A bio-guided study.

Currently, neurocysticercosis treatment involves two drugs: albendazole and praziquantel; however, their efficacy is suboptimal and new cysticidal drugs are needed. The present paper reports the cysticidal activity of extracts of the bark from Prunus serotina against Taenia crassiceps cysts and the isolation and identification of the main components of the most active extract. Results showed that all extracts displayed in vitro cysticidal activity (EC50=17.9-88.5µg/mL), being the methanolic the most active and selective. Also, methanolic extract exhibited in vivo efficacy at 300mg/kg which was similar to that obtained with albendazole. Bio-guided fractionation of methanolic extract led the isolation of 2,3-dihydro-5,7-dihydroxy-2-(4-hydroxyphenyl)-4H-1-benzopyran-4-one (naringenin, NGN), 3,4,5-trimethoxybenzoic acid and 1,3,5-trimethoxybenzene. NGN exhibited in vitro activity, in a time-concentration-dependent manner (EC50=89.3µM]. Furthermore, NGN at a dose of 376.1µmol/kg displayed similar in vivo efficacy than those obtained with albendazole at 188.4µmol/kg. NGN also caused a high level of damage in all parasite tissue in a similar manner than that observed with the methanolic extract. This study represents the first report of the cysticidal properties of the bark of P. serotina. NGN was identified as the main active compound of this specie and other studies are required to explore the potential of this flavanone as cysticidal agent.

Anti-inflammatory effect of Capuli cherry against LPS-induced cytotoxic damage in RAW 264.7 macrophages.

Capuli cherry (Prunus serotina Ehr. subsp. capuli (Cav.) McVaugh) fruits from the inter-Andean region of Ecuador were analysed to determine their bioactive compounds content, total antioxidant capacity, radical scavenging activity and their anti-inflammatory and protective effects against the cytotoxic damage mediated by lipopolysaccharide (LPS) in RAW 264.7 macrophages. Capuli fruits proved to be a natural source of bioactive compounds such as anthocyanins, vitamin C and ß-carotene as well as to present an important total antioxidant capacity and radical scavenging activities. RAW 264.7 macrophages were incubated with different concentration of Capuli crude extract and subsequently activated by LPS to determine the markers related to oxidative damage and the proinflammatory cytokine production. The markers of oxidative damage, nitrite levels, the interleukin 1ß messenger RNA levels and the tumor necrosis factor α mRNA levels and secretion were significantly decreased after the pre-incubated with Capuli extract and subsequently stimulated with LPS. In summary, Capuli extract attenuated the LPS-induced damage in RAW 264.7 macrophages due to its antioxidant and anti-inflammatory properties, showing that Capuli fruits may represent a relevant source of bioactive compounds with promising benefits for human health.

Incorporation of an invasive plant into a native insect herbivore food web.

The integration of invasive species into native food webs represent multifarious dynamics of ecological and evolutionary processes. We document incorporation of Prunus serotina (black cherry) into native insect food webs. We find that P. serotina harbours a herbivore community less dense but more diverse than its native relative, P. padus (bird cherry), with similar proportions of specialists and generalists. While herbivory on P. padus remained stable over the past century, that on P. serotina gradually doubled. We show that P. serotina may have evolved changes in investment in cyanogenic glycosides compared with its native range. In the leaf beetle Gonioctena quinquepunctata, recently shifted from native Sorbus aucuparia to P. serotina, we find divergent host preferences on Sorbus- versus Prunus-derived populations, and weak host-specific differentiation among 380 individuals genotyped for 119 SNP loci. We conclude that evolutionary processes may generate a specialized herbivore community on an invasive plant, allowing prognoses of reduced invasiveness over time. On the basis of the results presented here, we would like to caution that manual control might have the adverse effect of a slowing down of processes of adaptation, and a delay in the decline of the invasive character of P. serotina.

Seasonal Changes Affect Root Prunasin Concentration in Prunus serotina and Override Species Interactions between P. serotina and Quercus petraea.

The allocation of resources to chemical defense can decrease plant growth and photosynthesis. Prunasin is a cyanogenic glycoside known for its role in defense against herbivores and other plants. In the present study, fluctuations of prunasin concentrations in roots of Prunus serotina seedlings were hypothesized to be: (1) dependent on light, air temperature, and humidity; (2) affected by competition between Prunus serotina and Quercus petraea seedlings, with mulching with Prunus serotina leaves; (3) connected with optimal allocation of resources. For the first time, we determined prunasin concentration in roots on several occasions during the vegetative season. The results indicate that seasonal changes have more pronounced effects on prunasin concentration than light regime and interspecific competition. Prunus serotina invested more nitrogen in the synthesis of prunasin under highly restricted light conditions than in higher light environments. In full sun, prunasin in roots of Prunus serotina growing in a monoculture was correlated with growth and photosynthesis, whereas these relationships were not found when interspecific competition with mulching was a factor. The study demonstrates that prunasin concentration in Prunus serotina roots is the result of species-specific adaptation, light and temperature conditions, ontogenetic shift, and, to a lesser extent, interspecific plant-plant interactions.

Role of Nitric Oxide and Hydrogen Sulfide in the Vasodilator Effect of Ursolic Acid and Uvaol from Black Cherry Prunus serotina Fruits.

The present research aimed to isolate the non-polar secondary metabolites that produce the vasodilator effects induced by the dichloromethane extract of Prunus serotina (P. serotina) fruits and to determine whether the NO/cGMP and the H2S/KATP channel pathways are involved in their mechanism of action. A bioactivity-directed fractionation of the dichloromethane extract of P. serotina fruits led to the isolation of ursolic acid and uvaol as the main non-polar vasodilator compounds. These compounds showed significant relaxant effect on rat aortic rings in an endothelium- and concentration-dependent manner, which was inhibited by NG-nitro-L-arginine methyl ester (L-NAME), DL-propargylglycine (PAG) and glibenclamide (Gli). Additionally, both triterpenes increased NO and H2S production in aortic tissue. Molecular docking studies showed that ursolic acid and uvaol are able to bind to endothelial NOS and CSE with high affinity for residues that form the oligomeric interface of both enzymes. These results suggest that the vasodilator effect produced by ursolic acid and uvaol contained in P. serotina fruits, involves activation of the NO/cGMP and H2S/KATP channel pathways, possibly through direct activation of NOS and CSE.

Warming affects hatching time and early season survival of eastern tent caterpillars.

Climate change is disrupting species interactions by altering the timing of phenological events such as budburst for plants and hatching for insects. We combined field observations with laboratory manipulations to investigate the consequences of climate warming on the phenology and performance of the eastern tent caterpillar (Malacosoma americanum). We evaluated the effects of warmer winter and spring regimes on caterpillar hatching patterns and starvation endurance, traits likely to be under selection in populations experiencing phenological asynchrony, using individuals from two different populations (Washington, DC, and Roswell, GA). We also quantified the proximate and extended fitness effects of early food deprivation and recorded spring phenology of local caterpillars and their host plants. In addition, we conducted laboratory assays to determine if caterpillars are using plant chemical cues to fine-tune their hatching times. Warmer winter temperatures induced earlier hatching and caterpillars from GA survived starvation for periods that were 30% longer than caterpillars from DC. Warmer spring regimes reduced the starvation endurance of caterpillars overwintering in the wild but not in the laboratory. Early starvation dramatically reduced hatchling survival; however, surviving caterpillars did not show detrimental effects on pupal mass or development time. In the field, hatching preceded budburst in both 2013 and 2014 and the period of optimal foliage quality was 2 weeks shorter in 2013. Hatching time was unaffected by exposure to plant volatiles. Overall, we found that warmer temperatures can trigger late-season asynchrony by accelerating plant phenology and caterpillars from different populations exhibit differential abilities to cope with environmental unreliability.

Simultaneous quantification by HPLC of the phenolic compounds for the crude drug of Prunus serotina subsp. capuli.

CONTEXT: Prunus serotina Ehrenb. subsp. capuli (Cav.) McVaugh (Rosaceae), commonly known as "capulin", is a native North American tree, commercialized and used in folk medicine for the treatment of the hypertension, gastrointestinal illnesses, and cough. OBJECTIVE: This work developed a suitable HPLC method for quantifying the major active constituents of the infusion of P. serotina, the most important preparation consumed by populations around the world. MATERIALS AND METHODS: The analytical method was performed using a Fortis-RP column (150 mm × 4.6 mm; film thickness 5 µm). The mobile phase consisted of an isocratic acetate buffer solution (pH 2.7; A) and methanol (B) (65:35 v/v) at a flow rate of 1.0 mL min(-1). RESULTS: The proposed method was applied to the quantification of 1-3 in several samples of the leaves of P. serotina. The results indicated that amounts of 1-3 in the samples analyzed are uniform, and greater amounts of chlorogenic acid (2; 479.9 ± 33.6 µg g(-1), dry matter) along with hyperoside (1; 185.7 ± 55.3 µg g(-1), dry matter) were present. On the other hand, benzaldehyde (3; 118.2 ± 12.1 µg g(-1) dry matter) was found to be in lower concentration. CONCLUSIONS: A simple, sensitive, precise, and reproducible HPLC method for the simultaneous quantification of 1-3 in P. serotina was developed and validated. This is the first report on the quantification of 1-3 as active principles, and compound 1 was selected as a marker of P. serotina, which could be useful to guarantee the quality of the crude drug and herbal products.

Quantitative analysis of amygdalin and prunasin in Prunus serotina Ehrh. using (1) H-NMR spectroscopy.

INTRODUCTION: Prunus serotina is native to North America but has been invasively introduced in Europe since the seventeenth century. This plant contains cyanogenic glycosides that are believed to be related to its success as an invasive plant. For these compounds, chromatographic- or spectrometric-based (targeting on HCN hydrolysis) methods of analysis have been employed so far. However, the conventional methods require tedious preparation steps and a long measuring time. OBJECTIVE: To develop a fast and simple method to quantify the cyanogenic glycosides, amygdalin and prunasin in dried Prunus serotina leaves without any pre-purification steps using (1) H-NMR spectroscopy. METHODS: Extracts of Prunus serotina leaves using CH3 OH-d4 and KH2 PO4 buffer in D2 O (1:1) were quantitatively analysed for amygdalin and prunasin using (1) H-NMR spectroscopy. Different internal standards were evaluated for accuracy and stability. The purity of quantitated (1) H-NMR signals was evaluated using several two-dimensional NMR experiments. RESULTS: Trimethylsilylpropionic acid sodium salt-d4 proved most suitable as the internal standard for quantitative (1) H-NMR analysis. Two-dimensional J-resolved NMR was shown to be a useful tool to confirm the structures and to check for possible signal overlapping with the target signals for the quantitation. Twenty-two samples of P. serotina were subsequently quantitatively analysed for the cyanogenic glycosides prunasin and amygdalin. CONCLUSION: The NMR method offers a fast, high-throughput analysis of cyanogenic glycosides in dried leaves permitting simultaneous quantification and identification of prunasin and amygdalin in Prunus serotina.

Isolation and characterization of a TERMINAL FLOWER 1 homolog from Prunus serotina Ehrh.

Flowering control is one of the several strategies for gene containment of transgenic plants. TERMINAL FLOWER 1 (TFL1) is known to be involved in the transcriptional repression of genes for inflorescence development. Two TFL1 transcripts with different 3' UTR were cloned from black cherry (Prunus serotina Ehrh.) using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). Corresponding to the two TFL1 transcripts, two PsTFL1 gene sequences, 1248 bp and 1579 bp, were obtained and both contained the same 519 bp coding region which encoded a putative protein of 172 amino acid residues. The phylogenetic analysis of the amino acid sequences showed high identity of PsTFL1 to TFL1 orthologs of other Prunus species, including Yoshino cherry (Prunus × yedoensis Matsum.), peach (Prunus persica (L.) Batsch), apricot (Prunus armeniaca L.) and Japanese apricot (Prunus mume Sieb. et Zucc.). The real-time quantitative PCR detected a single copy of PsTFL1 gene sequences in the black cherry genome with two alleles. The gene expression of PsTFL1 was examined in several tissues including the stems, leaves, shoot tips, and vegetative and floral buds. The highest mRNA level was detected in shoot tips, and the lowest level in the leaves. Transgenic Arabidopsis thaliana (L.) Heynh. plants overexpressing PsTFL1 showed significantly delayed flowering. These plants also showed largely increased vegetative growth, plant height, number of nodes, trichome density, and the conversion of flower to shoot was observed at each node and shoot apex.

Variation in leaf and twig CO2 flux as a function of plant size: a comparison of seedlings, saplings and trees.

Rates of tissue-level function have been hypothesized to decline as trees grow older and larger, but relevant evidence to assess such changes remains limited, especially across a wide range of sizes from saplings to large trees. We measured functional traits of leaves and twigs of three cold-temperate deciduous tree species in Minnesota, USA, to assess how these vary with tree height. Individuals ranging from 0.13 to 20 m in height were sampled in both relatively open and closed canopy environments to minimize light differences as a potential driver of size-related differences in leaf and twig properties. We hypothesized that (H1) gas-exchange rates, tissue N concentration and leaf mass per unit area (LMA) would vary with tree size in a pattern reflecting declining function in taller trees, yet maintaining (H2) bivariate trait relations, common among species as characterized by the leaf economics spectrum. Taking these two ideas together yielded a third, integrated hypothesis that (H3) nitrogen (N) content and gas-exchange rates should decrease monotonically with tree size and LMA should increase. We observed increasing LMA and decreasing leaf and twig Rd with increasing size, which matched predictions from H1 and H3. However, opposite to our predictions, leaf and twig N generally increased with size, and thus had inverse relations with respiration, rather than the predicted positive relations. Two exceptions were area-based leaf N of Prunus serotina Ehrh. in gaps and mass-based leaf N of Quercus ellipsoidalis E. J. Hill in gaps, both of which showed qualitatively hump-shaped patterns. Finally, we observed hump-shaped relationships between photosynthetic capacity and tree height, not mirroring any of the other traits, except in the two cases highlighted above. Bivariate trait relations were weak intra-specifically, but were generally significant and positive for area-based traits using the pooled dataset. Results suggest that different traits vary with tree size in different ways that are not consistent with a universal shift towards a lower 'return on investment' strategy. Instead, species traits vary with size in patterns that likely reflect complex variation in water, light, nitrogen and carbon availability, storage and use.

Species recognition through wing interference patterns (WIPs) in Achrysocharoides Girault (Hymenoptera, Eulophidae) including two new species.

Wing interference patterns (WIPs) are shown to be an important tool for species recognition in the genus Achrysocharoides Girault (Hymenoptera: Eulophidae). This is demonstrated by combining information from two previously published papers, comprising two cases of cryptic species, and by new material including the description of two new species, Achrysocharoides maieri and Achrysocharoides serotinae from North America. The cryptic species were initially separated through their distinct male WIPs. Subsequent analyses of the external morphology uncovered additional morphological differences supporting the original findings through WIPs, and biological data further strengthened the identity of these species. The new species described here also differ in their WIPs but the WIPs are similar in both sexes. Thus they provide a strong link between male and female and demonstrate that WIPs can also be useful for species recognition when the sexes are otherwise difficult to associate. Both new species are from Connecticut, USA, and were reared from Phyllonorycter propinquinella (Braun) (Lepidoptera: Gracillariidae) on black cherry (Prunus serotina); Achrysocharoides maieri has also been reared from Ph. nr crataegella on pin cherry (Prunus pensylvanica). To facilitate the identification of the new species they are included in a previously published key to North American species of Achrysocharoides. As a supplement to colourful WIPs we also demonstrate that grey scale images of uncoated wings from scanning electron microscopy can be used for visualization of the thickness distribution pattern in wing membranes.