Flower power: Floral and resource manipulations reveal how and why reproductive trade‐offs occur for lowbush blueberry (Vaccinium angustifolium)

Plant reproductive trade‐offs are thought to be caused by resource limitations or other constraints, but more empirical support for these hypotheses would be welcome. Additionally, quantitative characterization of these trade‐offs, as well as consideration of whether they are linear, could yield additional insights. We expanded our flower removal research on lowbush blueberry (Vaccinium angustifolium) to explore the nature of and causes of its reproductive trade‐offs. We used fertilization, defoliation, positionally biased flower removal, and multiple flower removal levels to discern why reproductive trade‐offs occur in this taxon and to plot these trade‐offs along two continuous axes. We found evidence through defoliation that vegetative mass per stem may trade off with reproductive effort in lowbush blueberry because the two traits compete for limited carbon. Also, several traits including ripe fruit production per reproductive node and fruit titratable acidity may be “sink‐limited”—they decline with increasing reproductive effort because average reproductive structure quality declines. We found no evidence that reproductive trade‐offs were caused by nitrogen limitation. Use of reproductive nodes remaining per stem as a measure of reproductive effort indicated steeper trade‐offs than use of the proportion of nodes remaining. For five of six traits, we found evidence that the trade‐off could be concave down or up instead of strictly linear. Synthesis. To date, studies have aimed primarily at identifying plant reproductive trade‐offs. However, understanding how and why these trade‐offs occur represent the exciting and necessary next steps for this line of inquiry.     

Inhibition of matrix metalloproteinase activity in DU145 human prostate cancer cells by flavonoids from lowbush blueberry (Vaccinium angustifolium): possible roles for protein kinase C and mitogen-activated protein-kinase-mediated events

Regulation of the matrix metalloproteinases (MMPs) is crucial to regulate extracellular matrix (ECM) proteolysis which is important in metastasis. This study investigated the mechanism(s) by which three flavonoid-enriched fractions from lowbush blueberry (Vaccinium angustifolium) down-regulate MMP activity in DU145 human prostate cancer cells. Metalloproteinase activity was evaluated from cells exposed to "crude," anthocyanin-enriched (AN) and proanthocyanidin-enriched (PAC) fractions. Differential down-regulation of MMPs was observed. The activity of the endogenous tissue inhibitors of metalloproteinases (TIMPs) from these cells was also evaluated. Increases in TIMP-1 and TIMP-2 activity were observed in response to these fractions. The possible involvement of protein kinase C (PKC) and mitogen-activated protein (MAP) kinase pathways in the flavonoid-mediated decreases in MMP activity was observed. These findings indicate that blueberry flavonoids may use multiple mechanisms in down-regulating MMP activity in these cells.     

Safety and whole-body antioxidant potential of a novel anthocyanin-rich formulation of edible berries

Edible berry extracts rich in anthocyanins possess a broad spectrum of therapeutic, pharmacologic and anti-carcinogenic properties. Six berry extracts (wild blueberry, bilberry, cranberry, elderberry, raspberry seeds and strawberry), singly and in combination, were studied in our laboratories for antioxidant efficacy, cytotoxic potential, cellular uptake and anti-angiogenic properties. Combinations of edible berry extracts were evaluated to develop a synergistic formula, OptiBerry, which exhibited high oxygen radical absorbance capacity (ORAC) value, low cytotoxicity and superior anti-angiogenic properties compared to the other combinations tested. The current study sought to determine the broad spectrum safety and antioxidant potential of OptiBerry in vivo. Acute oral LD₅₀ of OptiBerry was greater than 5 g/kg in rats. Acute dermal LD₅₀ of OptiBerry was greater than 2 g/kg. No changes in the body weight or adverse effects were observed following necropsy. Primary skin and eye irritation studies were conducted in New Zealand albino rabbits. OptiBerry was classified as slightly irritating to the skin (primary skin irritation index 0.3) and minimally irritating to the eye (maximum mean total score 6.0). The antioxidant potential of OptiBerry was investigated in rats and mice by assessing GSH redox status in tissues as well as by a unique state-of-the-art electron paramagnetic resonance (EPR) imaging of whole-body redox status. A clinically relevant hyperbaric oxygen (HBO) exposure system (2 atm, 2 h) was employed to study the antioxidant properties of OptiBerry. OptiBerry feeding (8 weeks) significantly prevented HBO-induced GSH oxidation in the lung and liver of vitamin E-deficient Sprague Dawley rats. Furthermore, OptiBerry-fed mice, when exposed to HBO, demonstrated significant protection in whole-body HBO-induced oxidation compared to the unfed controls by EPR imaging. Taken together, these results indicate that OptiBerry is reasonably safe and possess antioxidant properties.     

Wild blueberry (Vaccinium angustifolium) consumption affects the composition and structure of glycosaminoglycans in Sprague-Dawley rat aorta

It has been documented that increased intake of polyphenols may provide protection against coronary heart disease and stroke. Blueberries (Vaccinium angustifolium) are one of the richest sources of antioxidants among fruits and vegetables. Phenolic compounds from berry extracts inhibit human low density lipoprotein and liposome oxidation. Glycosaminoglycans (GAGs) and proteoglycans (PGs) are structural components of aortas with great structural diversity. Their interaction with compounds such as enzymes, cytokines, growth factors, proteins and lipoproteins and their subsequent role in degenerative diseases has been documented. We investigated the effects of a diet rich in blueberries on the content and structure of GAGs. Sprague-Dawley rats were fed either a control (C) or a blueberry (B) diet for 13 weeks. Aortic tissue GAGs were isolated with papain digestion, alkaline borohydride treatment and anion-exchange chromatography. Cellulose acetate electrophoresis and treatment of the fractions with specific lyases revealed the presence of three GAG populations, i.e. hyaluronan (HA), heparan sulfate (HS) and galactosaminoglycans (GalAGs). Disaccharide composition was determined by high-performance capillary electrophoresis following enzymatic degradation. A 13% higher amount of total GAGs in aortas of B-fed rats was attributed to a higher content of GalAGs (67%). Determination of the sulfated disaccharides showed an overall lower concentration of oversulfated disaccharides in both HS and GalAG populations in the aortas of the B group. Our results demonstrate for the first time that a diet rich in blueberries results in structural alterations in rat aortic tissue GAGs. These changes may affect cellular signal transduction pathways and could have major consequences for the biological function of GAG molecules within the vascular environment.     

Seasonal acquisition of mineral nutrients by a chalcid gall on lowbush blueberry

Seasonal accumulation of various mineral nutrients by galls of the chalcidHemadas nubilipennis on lowbush blueberry were examined in various phases of gall development. Levels of copper, nickel iron and zinc were higher in tissues of galls in initiation phase compared to tips of ungalled shoots and in general were higher than galls in growth and maturation phases. Levels of manganese and magnesium in gall tissues decrease throughout the season compared to ungalled shoots. Levels of calcium are less in gall tissue in initiation phase compared to ungalled shoots, and elevated in maturation phase. Patterns of mineral concentration within galls at various phases of development are related to known physiological roles of minerals in plant nutrition; however we conclude that patterns are variable and their significance in gall physiology requires further study.     

Wild blueberry (Vaccinium angustifolium) consumption improves inflammatory status in the obese Zucker rat model of the metabolic syndrome

The metabolic syndrome (MetS) is a major public health problem in the United States. Chronic inflammation is a critical component of the MetS, leading to dramatically increased risk of type II diabetes and cardiovascular disease.This study investigates the ability of a wild-blueberry-enriched diet to improve the proinflammatory status associated with MetS in the obese Zucker rat (OZR). Circulating levels of key inflammatory markers and their expression in the liver and abdominal adipose tissue were examined in OZR and its genetic control, the lean Zucker rat (LZR), after feeding a control or an 8% wild blueberry diet (WB) for 8 weeks from age 8 to 16 weeks.In the OZR, WB consumption resulted in decreased plasma concentrations of tumor necrosis factor (TNF)-α (?25.6%, P<.05), interleukin (IL)-6 (?14.9%, P<.05) and C-reactive protein (CRP) (?13.1%, P<.05) and increased adiponectin concentration (+21.8%, P<.05). Furthermore, expression of IL-6, TNF-α and nuclear factor (NF)-kB was down-regulated in both the liver (?65%, ?59% and ?25%, respectively) and the abdominal adipose tissue (?64%, ?52% and ?65%), while CRP expression was down-regulated only in the liver (?25%). In the abdominal adipose tissue, similar trends were also observed in LZR following WB treatment, with decreased liver expression of NF-kB, CRP, IL-6 and TNF-α (?24%, ?16%, ?21% and ?50%) and increased adiponectin expression (+25%).Results of this study suggest that wild blueberry consumption exerts an overall anti-inflammatory effect in the OZR, a model of the metabolic syndrome.     

The effects of pruning,fertilizers, and organic amendments on lowbush blueberry production

A four-year field study was conducted on a Hebert gravelly sandy loam (pH 4.5) in Nova Scotia to assess the effects of pruning management and seven fertility amendments on lowbush blueberry (Vaccinium augustifolium Ait) production (yield, above ground and root tissue composition) and soil fertility. Pruning by oil burning produced higher fruit yields than flail mowing but burning had the opposite effect on the plant N content (with a lesser influence on above ground Mn and Zn). None of the fertility treatments (chicken manure, dairy manure, swine manure, urea, sawdust, NPK, NPK+S+Lime+Micronutrients) produced fruit yields significantly greater than the control. Treatments provided the equivalent of 50kg total N/ha/2-yr cycle. Treatments influenced tissue N, P, K, Ca, Mg, B, Mn, Cu, Zn and Mo levels. In general, the three manure treatments produced the highest levels of plant macronutrients; the urea treatment produced the lowest levels of plant nutrients. In most cases, extractable levels of soil P, K, Ca and Mg were highly correlated with the plant tissue content of these elements. Overall, the dairy manure treated soils were the highest in soil fertility.     

Occurrence of unreduced pollen in diploid blueberry species,Vaccinium sect. Cyanococcus

Summary A total of 1475 individuals belonging to 43 natural populations of seven diploid (2x) blueberry species (Vaccinium section Cyanococcus) and two natural interspecific 2x hybrid populations were evaluated for unreduced pollen production. Significant differences were found in the frequency of unreduced pollen producers between species and within and between populations of the same species. Individuals with 1% or more unreduced pollen were considered unreduced pollen producers. The average frequency of unreduced pollen producers in these diploid species was 13.5%, ranging from 7.4% (V. corymbosum) to 18.4% (V. darrowii). The frequency of unreduced pollen grains in individual clones varied from <-1% to 28.6%. The production of unreduced pollen was not associated with male fertility. The widespread occurrence of unreduced pollen in the diploid species should allow the introgression of this germ plasm to the tetraploid level via unilateral sexual polyploidization.New Jersey Agricultural Experiment Station, Publication No. D-12163-18-91, supported by State and U.S. Federal funds, USDA-ARS Specific Cooperative Agreement No. 58-3615-9-068     

Development of a novel in vitro rooting culture system for the micropropagation of highbush blueberry (Vaccinium corymbosum) seedlings

Plant Cell, Tissue and Organ Culture (PCTOC) - Here, we established a novel, efficient, and cost-effective rooting culture system for highbush blueberry (Vaccinium corymbosum). This method...     

Protective effect of anthocyanin-rich extract from bilberry (Vaccinium myrtillus L.) against myelotoxicity induced by 5-fluorouracil

The toxicities associated with 5-fluorouracil (5-FU), a potent broad-spectrum chemotherapeutic agent, can not only affect the morbidity and the efficacy of chemotherapy but also limit its clinical use. The objective of this study is to investigate the effects of a commercial anthocyanin-rich extract from bilberry (AREB) against 5-FU-induced myelotoxicity in vivo, and against chemosensitivity to 5-FU in vitro. A single injection of 5-FU at 200 mg/kg induced severe peripheral erythrocytopenia, thrombocytopenia and leucopenia as well as hypocellularity of the spleen and bone marrow in C57BL/6 mice. Oral administration of 500 mg/kg of AREB for 10 days significantly increased the number of red blood cells, neutrophils, and monocytes in peripheral blood to 1.2-fold, 9-fold, and 6-fold, respectively, compared with those seen after treatment with 5-FU alone (p< 0.05-0.001). The hypocellularity of the spleen and bone marrow caused by 5-FU was also distinctly alleviated in the AREB-treated group. Furthermore, AREB treatment with 50 and 100 microg/ml as a monomeric anthocyanin did not interfere with, but rather enhanced the chemotherapeutic efficacy of 5-FU in vitro. These results suggest that AREB may have protective potential against 5-FU-induced myelotoxiciy and/or the ability to enhance the chemotherapeutic effectiveness of 5-FU.     

Griseofulvin-producing Xylaria endophytes of Pinus strobus and Vaccinium angustifolium: evidence for a conifer-understory species endophyte ecology

During three decades of research on conifer endophytes of the Acadian forest, numerous insights have been gained in conifer-fungal ecology and secondary metabolite production. Recently, we have explored endophyte assemblages of understory plants commonly occurring with pine. Here we report for the first time the production of the potent antifungal compound griseofulvin by a fungal endophyte isolated from eastern white pine (Pinus strobus) needles and lowbush blueberry (Vaccinium angustifolium) stems from the Acadian forest of New Brunswick and Nova Scotia, Canada. Maximum likelihood phylogenetic analysis has placed the endophyte strains as an undescribed Xylaria sp. Our study highlights the complexity of endophyte-host lifecycles and points to the existence of a pine-blueberry ecotype. Aside from griseofulvin, piliformic acid was isolated from one of the pine endophytes. This compound has been reported from Xylaria and related species but not from Penicillium species known to produce griseofulvin.     

Hypoglycemic activities of MTP-3631, a novel thiopyranopyrimidine derivative.

MTP-3631 is a novel thiopyranopyrimidine derivative structurally different from any existent hypoglycemic agents. MTP-3631 markedly decreased basal blood glucose and improved glucose intolerance in genetically diabetic C57BL/6J ob/ob mice, which was not affected by tolbutamide. MTP-3631 also caused hypoglycemic effects in normal rats, but no change was observed in plasma insulin level. Unlike buformin, MTP-3631 did not change plasma lactate level in ob/ob mice. These results suggest that the hypoglycemic mechanism of MTP-3631 may be essentially different from those of sulfonylureas and biguanides.     

Contextualising pollination benefits: effect of insecticide and fungicide use on fruit set and weight from bee pollination in lowbush blueberry

Current approaches to determining the value of insect pollinators to crop yield assume that plants are primarily pollen limited. This is particularly relevant in a crop such as lowbush blueberry, Vaccinium angustifolium, where no fruit will set without insect‐mediated cross‐pollination. However, such valuations usually ignore other factors that are necessary to maximise crop yields. We conducted an experiment to test whether yields of lowbush blueberry attributed to pollinator activity increased independently of pest management. The experiment was a 2 × 2 factorial design, incorporating two intensities of pollination (25% or 100% of flowers), and two levels of insect and disease management with recommended fungicide and insecticide sprays (‘full inputs’ or ‘no inputs’). We demonstrated an interaction between these two factors, such that increased fruit set at harvest was only possible if 100% pollination was combined with the ‘full input’ treatment. Furthermore, increases in fruit weight among the remaining treatments were only realised in the ‘full input’ plots. These results suggest that the value accorded to pollinator activity in blueberries is strongly dependent upon pest and disease management of the crop.     

RAPD-based genetic linkage map of blueberry derived from a cross between diploid species (Vaccinium darrowi and V. elliottii)

An initial genetic linkage map for blueberry has been constructed from over 70 random amplified polymorphic DNA (RAPD) markers that segregated 11 in a testcross population of 38 plants. The mapping population was derived from a cross between two diploid blueberry plants: the F₁ interspecific hybrid (Vaccinium darrowi Camp x V. elliottii Chapm.) and another V. darrowi plant. The map currently comprises 12 linkage groups (in agreement with the basic blueberry chromosome number) and covers a total genetic distance of over 950cM, with a range of 3–30cM between adjacent markers. The use of such a map for identifying molecular markers linked to genes controlling chilling requirement and cold hardiness is discussed.     

Preparative separation of chlorogenic acid by centrifugal partition chromatography from highbush blueberry leaves (Vaccinium corymbosum L.)

Introduction – Blueberries (genus Vaccinium) have gained worldwide focus because of the high anthocyanin content of their fruits. In contrast, the leaves of blueberry have not attracted any attention, even though they contain large quantities of chlorogenic acid, a strong antioxidant compound. Objective – The aim of this investigation was the quantification and preparative isolation of chlorogenic acid (5‐caffeoylquinic acid, 5‐CQA) from blueberry leaves using a new separation scheme, centrifugal partition chromatography (CPC). Methodology – A water fraction containing a high concentration of 5‐CQA (14.5% of dry weight extract) was obtained by defatting a crude methanol extract from blueberry leaves. CPC was applied to isolate 5‐CQA from this water fraction using a two‐phase solvent system of ethyl acetate–ethanol–water at a volume ratio 4:1:5 (v/v/v). The flow‐rate of mobile phase was 2 mL/min with the ascending mode while rotating at 1200 rpm. The eluate was monitored at 330 nm. The structure of chlorogenic acid in the CPC fraction was confirmed with HPLC, UV, ESI/MS and NMR spectra. Results – The HPLC chromatogram showed that the fractions collected by CPC contained chlorogenic acid with 96% purity based on peak area percentage. The total amount of chlorogenic acid isolated from 0.5 g of a water fraction was 52.9 mg, corresponding to 10.6% of the water fraction. The isolated compound was identified successively as 5‐CQA with MS (parent ion at m/z 355.1 [M + H]⁺) and ¹H NMR spectra [caffeoyl moiety in the down field (δ 6.0–8.0 ppm) and quinic acid moiety in the up field (δ 2.0–5.5 ppm)]. Conclusion – 5‐CQA was successfully isolated from blueberry leaves by the CPC method in a one‐step procedure, indicating a further potential use for blueberry leaves. Copyright © 2010 John Wiley & Sons, Ltd.     

In vitro antimicrobial activity of a novel propolis formulation (Actichelated propolis)

AIMS: This study compared in vitro activities of Actichelated propolis (a multicomposite material obtained with mechano-chemichal activation) and of a hydroalcoholic extract of propolis. METHODS AND RESULTS: Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC), determined by means of microdilution broth method, against five strains of Staphylococcus aureus, Streptococcus pyogenes, Haemophilus influenzae, Enterococcus spp., Escherichia coli, Proteus mirabilis and Pseudomonas aeruginosa, showed a greater potency of Actichelated propolis (MIC range: 0.016-4 mg flavonoids ml(-1)) in respect to the hydroalcoholic extract (MIC range: 0.08-21.4 mg flavonoids ml(-1)). Concentrations of Actichelated propolis active against adenovirus, influenza virus, parainfluenza virus and herpes virus type 1 were at least 10 times lower than those of the hydroalcoholic extract. Preincubation of Strep. pyogenes and H. influenzae with subinhibitory concentrations of Actichelated propolis (1/4 and 1/8 x MIC) significantly reduced the number of bacteria that adhered to human buccal cells. CONCLUSIONS: Actichelated propolis has proven to possess antibacterial and antiviral activity higher than a hydroalcoholic extract, being also able to interfere on bacterial adhesion to human oral cells. SIGNIFICANCE AND IMPACT OF THE STUDY: This new formulation of propolis showing better antimicrobial and physical characteristics could improve the application of propolis in respiratory tract infections.     

Identification of dehydrin-like proteins responsive to chilling in floral buds of blueberry (Vaccinium, section Cyanococcus).

The level of three major polypeptides of 65, 60, and 14 kD increased in response to chilling unit accumulation in floral buds of a woody perennial, blueberry (Vaccinium, section Cynaococcus). The level of the polypeptides increased most dramatically within 300 h of chilling and decreased to the prechilling level with the initiation of budbreak. Cold-hardiness levels were assessed for dormant buds of Vaccinium corymbosum and Vaccinium ashei after different chilling treatments until the resumption of growth. These levels coincided with the level of the chilling-responsive polypeptides. Like some other previously described cold-induced proteins in annual plants, the level of the chilling-induced polypeptides also increased in leaves in response to cold treatment; the chilling-induced polypeptides were heat stable, resisting aggregation after incubation at 95 degrees C for 15 min. By fractionating bud proteins first by isoelectric point (pI) and then by molecular mass, the pI values of the 65- and 60-kD polypeptides were found to be 7.5 to 8.0 and the pI value of the 14-kD polypeptide was judged to be 8.5. Purification of the 65- and 60-kD polypeptides, followed by digestion with endoproteinase Lys-C and sequencing of selected fragments, revealed similarities in amino acid composition between the 65- and 60-kD polypeptides and dehydrins. Indeed, antiserum to the lysine-rich consensus sequence EKKGIMDKIKEKLPG of dehydrin proteins cross-reacted to all three of the major chilling-responsive polypeptides of blueberry, identifying these as dehydrins or dehydrin-like proteins.     

Paphiopedilum angustifolium,A New Species of Orchidaceae from Vietnam

Paphiopedilum angustifolium R.F.Guo et Z.J.Liu,sp.nov.Fig.1:1-8.Species nova Paphiopedilo puberulo S.P.Lei et J.Y.Zhang affinis,a quo differt staminodio ovato-cordato glabro viridulo praeter centrum vitta una lata alba et ab ejus latere alterutro alia viridi-fusca praeditum.     

狭叶兜兰,越南兰科一新种

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Hypoglycemic effect of 13-membered ring thiocyclitol, a novel alpha-glucosidase inhibitor from Kothala-himbutu (Salacia reticulata)

A novel 13-membered ring thiocyclitol, isolated from an aqueous extract of Kothala-himbutu (Salacia reticulata), inhibited alpha-glucosidase in vitro. The inhibitory activity was investigated by maltose- and sucrose-loading on Wistar rats. This study found significant lowering of postprandial glucose levels, and the potency of 13-membered ring thiocyclitol was confirmed in vivo.