Research Update:Prunus serotina,Cherry Bark,Cherry Fruit.

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   Phytochemical info of Prunus serotina,Cherry Bark,Cherry Fruit.

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   Research Update:Prunus serotina,Cherry Bark,Cherry Fruit.

  Effects of different pretreatments on germination of Prunus serotina seed sources.:J Environ Biol. 2007 Jan;28(1):99-104.Esen D, Yildiz O, Sarginci M, Isik K.Abant Izzet Baysal University, Duzce Forestry Faculty, Konuralp Yerleskesi, Konuralp, Duzce-81620, Turkey. guzelfethiye@yahoo.com

 Establishing intensive plantations of fast growing hardwood tree species that have high market values in the forest industry can narrow the gap between Turkey's demand and the supply of quality hardwood products. Black cherry (P. serotina Ehrh.) is a fast growing hardwood species with a high market value. Introducing and intensively growing black cherry (BC) in Turkey may significantly reduce the country's quality wood shortage. Adequate seed germination constitutes the first essential step for successful establishments. In this paper effects of different pretreatments, including artificial and natural stratification, on the seeds of different BC seed sources (SSs) were studied. Pretreatments had substantial effects on the dormancy breaking and germination behaviours of the SSs. Consecutive periods of complex warm and cold artificial stratification regimes longer than 90 days or natural stratification (where seeds were assumed to be naturally exposed to this complexity) resulted in best dormancy breaking and, in turn, germination among all pretreatments. Deeper dormancy and reduced germination rates of some BC seeds as the altitude of the source increases might suggest an ecological adaptive strategy of the species. BC may have deeper morphophysiological dormancy than is commonly believed. Seed size may have a positive effect on seed germination.

  Ozone uptake (flux) as it relates to ozone-induced foliar symptoms of Prunus serotina and Populus maximowiziixtrichocarpa.:Environ Pollut. 2007 May 22;Orendovici-Best T, Skelly JM, Davis DD, Ferdinand JA, Savage JE, Stevenson RE.School of Forest Resources, The Pennsylvania State University, University Park, PA 16802, USA.

 Field studies were conducted during 2003 and 2004 from early June to the end of August, at 20 sites of lower or higher elevation within north-central Pennsylvania, using seedlings of black cherry (Prunus serotina, Ehrh.) and ramets of hybrid poplar (Populus maximowiziixtrichocarpa). A linear model was developed to estimate the influence of local environmental conditions on stomatal conductance. The most significant factors explaining stomatal variance were tree species, air temperature, leaf vapor pressure deficit, elevation, and time of day. Overall, environmental factors explained less than 35% of the variation in stomatal conductance. Ozone did not affect gas exchange rates in either poplar or cherry. Ozone-induced foliar injury was positively correlated with cumulative ozone exposures, expressed as SUM40. Overall, the amount of foliar injury was better correlated to a flux-based approach rather than to an exposure-based approach. More severe foliar injuries were observed on plants growing at higher elevations.

  Photosynthetic responses to understory shade and elevated carbon dioxide concentration in four northern hardwood tree species.:Tree Physiol. 2006 Dec;26(12):1589-99.Sefcik LT, Zak DR, Ellsworth DS.School of Natural Resources and Environment, 440 Church Street, University of Michigan, Ann Arbor, MI 48109-1041, USA. ltsefcik@umich.edu

 Seedling responses to elevated atmospheric CO(2) concentration ([CO(2)]) and solar irradiance were measured over two growing seasons in shade-tolerant Acer saccharum Marsh. and Fagus grandifolia J.F. Ehrh. and shade-intolerant Prunus serotina, a J.F. Ehrh. and Betula papyrifera Marsh. Seedlings were exposed to a factorial combination of [CO2] (ambient and elevated (658 micromol mol-1)) and understory shade (deep and moderate) in open-top chambers placed in a forest understory. The elevated [CO(2)] treatment increased mean light-saturated net photosynthetic rate by 63% in the shade-tolerant species and 67% in the shade-intolerant species. However, when measured at the elevated [CO(2)], long-term enhancement of photosynthesis was 10% lower than the instantaneous enhancement seen in ambient-[CO(2)]-grown plants (P < 0.021). Overall, growth light environment affected long-term photosynthetic enhancement by elevated [CO(2)]: as the growth irradiance increased, proportional enhancement due to elevated [CO(2)] decreased from 97% for plants grown in deep shade to 47% for plants grown in moderate shade. Results suggest that in N-limited northern temperate forests, trees grown in deep shade may display greater photosynthetic gains from a CO(2)-enriched atmosphere than trees growing in more moderate shade, because of greater downregulation in the latter environment. If photosynthetic gains by deep-shade-grown plants in response to elevated [CO(2)] translate into improved growth and survival of shade-intolerant species, it could alter the future composition and dynamics of successional forest communities.

  High-performance liquid chromatographic identification of flavonoid monoglycosides from Prunus serotina ehrh.:Acta Pol Pharm. 2005 Nov-Dec;62(6):435-41.Olszewska M.Department of Pharmacognosy, Faculty of Pharmacy, Medical University of L¨®d?, 1 Muszy¨½iski St., 90-151 L¨®d?, Poland. molszewska@pharm.am.lodz.pl

 Five minor flavonoid monosides, glycosides of quercetin and kaempferol, together with three previously isolated compounds were identified cochromatographically in P. serotina Ehrh. leaves and flowers (inflorescences) using RP-HPLC and TLC techniques and finally determined as quercetin 3-O-alpha-L-arabinofuranoside (avicularin), 3-O-alpha-L-arabinopyranoside (guaijaverin), 3-O-beta-D-xylopyranoside (reynoutrin), 3-O-beta-D-glucopyranoside (isoquercitrin), 3-O-beta-D-galactopyranoside (hyperoside) followed by kaempferol 3-O-alpha-L-arabinofuranoside (juglanin), 3-O-beta-D-xylopyranoside and 3-O-beta-D-glucopyranoside (astragalin). Moreover, two further minor flavonols were isolated from the leaves, characterized by hydrolysis experiments, UV and 1H NMR spectroscopy, and identified finally as isorhamnetin 3-O-alpha-arabinofuranoside and isorhamnetin 3-O-beta-xylopyranoside, the rare natural products.

  Crystal macropattern development in Prunus serotina (Rosaceae, Prunoideae) leaves.:Ann Bot (Lond). 2006 May;97(5):723-9. Epub 2006 Mar 2.Lersten NR, Horner HT.Department of Ecology, Evolution and Organismal Biology, Iowa State University, Ames, IA 50011, USA.

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


  Field responses of Prunus serotina and Asclepias syriaca to ozone around southern Lake Michigan.:Environ Pollut. 2006 Jul;142(2):354-66. Epub 2005 Dec 15.Bennett JP, Jepsen EA, Roth JA.U.S. Geological Survey and Institute for Environmental Studies, University of Wisconsin, Madison, WI 53706, USA. jpbennet@wisc.edu

 Higher ozone concentrations east of southern Lake Michigan compared to west of the lake were used to test hypotheses about injury and growth effects on two plant species. We measured approximately 1000 black cherry trees and over 3000 milkweed stems from 1999 to 2001 for this purpose. Black cherry branch elongation and milkweed growth and pod formation were significantly higher west of Lake Michigan while ozone injury was greater east of Lake Michigan. Using classification and regression tree (CART) analyses we determined that departures from normal precipitation, soil nitrogen and ozone exposure/peak hourly concentrations were the most important variables affecting cherry branch elongation, and milkweed stem height and pod formation. The effects of ozone were not consistently comparable with the effects of soil nutrients, weather, insect or disease injury, and depended on species. Ozone SUM06 exposures greater than 13 ppm-h decreased cherry branch elongation 18%; peak 1-h exposures greater than 93 ppb reduced milkweed stem height 13%; and peak 1-h concentrations greater than 98 ppb reduced pod formation 11% in milkweed.

  Anti-proliferative effect of horehound leaf and wild cherry bark extracts on human colorectal cancer cells.:Oncol Rep. 2006 Jan;15(1):275-81.Yamaguchi K, Liggett JL, Kim NC, Baek SJ.Laboratory of Environmental Carcinogenesis, Department of Pathobiology, College of Veterinary Medicine, University of Tennessee, Knoxville, 37996, USA.

 Marubium vulgare (horehound) and Prunus serotina (wild cherry) have been traditionally used for the treatment of inflammatory-related symptoms such as cold, fever, and sore throat. In this report, we show that extracts of anti-inflammatory horehound leaves and wild cherry bark exhibit anti-proliferative activity in human colorectal cancer cells. Both horehound and wild cherry extracts cause suppression of cell growth as well as induction of apoptosis. We found that horehound extract up-regulates pro-apoptotic non-steroidal anti-inflammatory drug-activated gene (NAG-1) through transactivation of the NAG-1 promoter. In contrast, wild cherry extract decreased cyclin D1 expression and increased NAG-1 expression in HCT-116 and SW480 cell lines. Treatment with wild cherry extract resulted in the suppression of beta-catenin/T cell factor transcription, as assessed by TOP/FOP reporter constructs, suggesting that suppressed beta-catenin signaling by wild cherry extract leads to the reduction of cyclin D1 expression. Our data suggest the mechanisms by which these extracts suppress cell growth and induce apoptosis involve enhanced NAG-1 expression and/or down-regulation of beta-catenin signaling, followed by reduced cyclin D1 expression in human colorectal cancer cells. These findings may provide mechanisms for traditional anti-inflammatory products as cancer chemopreventive agents.

  Flavonoids from Prunus serotina Ehrh.:Acta Pol Pharm. 2005 Mar-Apr;62(2):127-33.Olszewska M.Department of Pharmacognosy, Faculty of Pharmacy, Medical University of L¨®d?, 1 Muszy¨½ski Str, 90-151 L¨®d?, Poland. molszewska@pharm.am.lodz.pl

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

  Prunus spp. intoxication in ruminants: a case in a goat and diagnosis by identification of leaf fragments in rumen contents.:J Vet Diagn Invest. 2004 Nov;16(6):593-9.Radi ZA, Styer EL, Thompson LJ.Tifton Diagnostic and Investigational Laboratory, College of Veterinary Medicine, The University of Georgia, Tifton, GA 31793-1389, USA.

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

  Element accumulation patterns of deciduous and evergreen tree seedlings on acid soils: implications for sensitivity to manganese toxicity.:Tree Physiol. 2005 Jan;25(1):85-92.St Clair SB, Lynch JP.Intercollegiate Graduate Program in Ecological and Molecular Plant Physiology, The Pennsylvania State University, 102 Tyson Building, University Park, PA 16802, USA.

 Foliar nutrient imbalances, including the hyperaccumulation of manganese (Mn), are correlated with symptoms of declining health in sensitive tree species growing on acidic forest soils. The objectives of this study were to: (1) compare foliar nutrient accumulation patterns of six deciduous (sugar maple (Acer saccharum Marsh.), red maple (Acer rubrum L.), red oak (Quercus rubra L.), white oak (Quercus alba L.), black cherry (Prunus serotina Ehrh.) and white ash (Fraxinus americana L.)) and three evergreen (eastern hemlock (Tsuga canadensis L.), white pine (Pinus strobus L.) and white spruce (Picea glauca (Moench) Voss.)) tree species growing on acidic forest soils; and (2) examine how leaf phenology and other traits that distinguish evergreen and deciduous tree species influence foliar Mn accumulation rates and sensitivity to excess Mn. For the first objective, leaf samples of seedlings from five acidic, non-glaciated field sites on Pennsylvania's Allegheny Plateau were collected and analyzed for leaf element concentrations. In a second study, we examined growth and photosynthetic responses of seedlings exposed to excess Mn in sand culture. In field samples, Mn in deciduous foliage hyperaccumulated to concentrations more than twice as high as those found in evergreen needles. Among species, sugar maple was the most sensitive to excess Mn based on growth and photosynthetic measurements. Photosynthesis in red maple and red oak was also sensitive to excess Mn, whereas white oak, black cherry, white ash and the three evergreen species were tolerant of excess Mn. Among the nine species, relative rates of photosynthesis were negatively correlated with foliar Mn concentrations, suggesting that photosynthetic sensitivity to Mn is a function of its rate of accumulation in seedling foliage.


  Physiological and foliar symptom response in the crowns of Prunus serotina, Fraxinus americana and Acer rubrum canopy trees to ambient ozone under forest conditions.:Environ Pollut. 2005 Feb;133(3):553-67.Schaub M, Skelly JM, Zhang JW, Ferdinand JA, Savage JE, Stevenson RE, Davis DD, Steiner KC.Graduate Ecology Program, Pennsylvania State University, University Park, PA 16802, USA.

 The crowns of five canopy dominant black cherry (Prunus serotina Ehrh.), five white ash (Fraxinus americana L.), and six red maple (Acer rubrum L.) trees on naturally differing environmental conditions were accessed with scaffold towers within a mixed hardwood forest stand in central Pennsylvania. Ambient ozone concentrations, meteorological parameters, leaf gas exchange and leaf water potential were measured at the sites during the growing seasons of 1998 and 1999. Visible ozone-induced foliar injury was assessed on leaves within the upper and lower crown branches of each tree. Ambient ozone exposures were sufficient to induce typical symptoms on cherry (0-5% total affected leaf area, LAA), whereas foliar injury was not observed on ash or maple. There was a positive correlation between increasing cumulative ozone uptake (U) and increasing percent of LAA for cherry grown under drier site conditions. The lower crown leaves of cherry showed more severe foliar injury than the upper crown leaves. No significant differences in predawn leaf water potential (psi(L)) were detected for all three species indicating no differing soil moisture conditions across the sites. Significant variation in stomatal conductance for water vapor (g(wv)) was found among species, soil moisture, time of day and sample date. When comparing cumulative ozone uptake and decreased photosynthetic activity (P(n)), red maple was the only species to show higher gas exchange under mesic vs. drier soil conditions (P < 0.05). The inconsistent differences in gas exchange response within the same crowns of ash and the uncoupling relationship between g(wv) and P(n) demonstrate the strong influence of heterogeneous environmental conditions within forest canopies.

  Foliar response of black cherry (Prunus serotina) clones to ambient ozone exposure in central Pennsylvania.:Environ Pollut. 1999 Jun;105(3):325-31.Lee JC, Skelly JM, Steiner KC, Zhang JW, Savage JE.School of Forest Resources, 220 Forest Resource Laboratory, The Pennsylvania State University, University Park, PA 16802, USA.

 During late summer of 1996 and 1997 we examined ozone-induced foliar injury in a plantation of 111 black cherry trees (ramets) comprising 15 clones originating from wild ortets growing in the Allegheny National Forest, Pennsylvania, and the Monongahela National Forest, West Virginia. The experimental plantation was a clonal seed orchard in Centre County, Pennsylvania, started in 1971 using ortet buds grafted onto seedling rootstocks of mixed origin. Clones differed significantly in severity of foliar injury symptoms (F=31.83, p<0.001). One clone (R-12) had significantly more foliar injury with >50% leaf area affected than other clones during both years. In contrast, clone R-14, which is from the same area in northcentral Pennsylvania as R-12, exhibited significantly less injury (LAA<6%). Although ambient O(3) concentrations were similar in both years, foliar injury was significantly greater (15.7%) in 1996 than in 1997 (9.9%). This is probably explained by lower stomatal conductance in 1997 caused by drier and hotter weather patterns in June and July of that year. Despite very different weather patterns and overall levels of injury in 1996 and 1997, mean clonal injury was significantly correlated between both years of assessment (r=0.92, p<0.001). Within tree crowns, foliage in lower and inner crown positions was significantly more injured than foliage in upper and exterior crown positions. There was no evidence of geographically based population differences in sensitivity to foliar O(3) injury. On the contrary, results demonstrate that wild genotypes of proximal geographic origin may differ greatly in sensitivity.

  Interactive effects of ozone and elevated carbon dioxide on the growth and physiology of black cherry, green ash, and yellow-poplar seedlings.:Environ Pollut. 1999 Aug;106(2):237-48.Loats KV, Rebbeck J.Biology Department, Denison University, Granville, OH 43023, USA.

 Potted seedlings of black cherry (Prunus serotina Ehrh.) (BC), green ash (Fraxinus pennsylvanica Marsh.) (GA), and yellow-poplar (Liriodendron tulipifera L.) (YP) were exposed to one of the four treatments: (1) charcoal-filtered air (CF) at ambient CO(2) (control); (2) twice ambient O(3) (2 x O(3)); (3) twice ambient CO(2) (650 microl l(-1)) plus CF air (2 x CO(2)); or (4) twice ambient CO(2) (650 microl l(-1)) plus twice ambient O(3) (2 x CO(2) + 2 x O(3)). The treatments were duplicated in eight continuously stirred tank reactors for 10 weeks. Gas exchange was measured during the last 3 weeks of treatment and all seedlings were destructively harvested after 10 weeks. Significant interactive effects of O(3) and CO(2) on the gas exchange of all three species were limited. The effects of elevated CO(2) and O(3), singly and combined, on light-saturated net photosynthesis (A(max)) and stomatal conductance (g(s)) were inconsistent across species. In all three species, elevated O(3) had no effect on g(s). Elevated CO(2) significantly increased A(max) in GA and YP foliage, and decreased g(s) in YP foliage. Maximum carbon exchange rates and quantum efficiencies derived from light-response curves increased, while compensation irradiance and dark respiration decreased in all three species when exposed to 2 x CO(2). Elevated O(3) affected few of these parameters but any change that was observed was opposite to that from exposure to 2 x CO(2)-air. Interactive effects of CO(2) and O(3) on light-response parameters were limited. Carboxylation efficiencies, derived from CO(2)-response curves (A/C(i) curves) decreased only in YP foliage exposed to 2 x CO(2)-air. In general, growth was significantly stimulated by 2 x CO(2) in all three species; though there were few significant growth responses following exposure to 2 x O(3) or the combination of 2 x CO(2) plus 2 x O(3). Results indicate that responses to interacting stressors such as O(3) and CO(2) are species specific.

  Foliar injury, leaf gas exchange and biomass responses of black cherry (Prunus serotina Ehrh.) half-sibling families to ozone exposure.:Environ Pollut. 2000 Jan;107(1):117-26.Kouterick KB, Skelly JM, Fredericksen TS, Steiner KC, Kolb TE, Ferdinand JA.Department of Plant Pathology, The Pennsylvania State University, University Park, PA 16802, USA.

 Open pollinated families of black cherry seedlings were studied to determine genotypic differences in foliar ozone injury and leaf gas exchange in 1994 and growth response following three growing seasons. An O(3)-sensitive half-sibling family (R-12) and an O(3)-tolerant half-sibling family (MO-7) planted in natural soil were studied along with generic nursery stock (NS) seedlings. Ozone exposure treatments were provided through open top chambers and consisted of 50, 75, and 97% of ambient ozone, and open plots from May 9 to August 26, 1994. Ambient ozone concentrations reached an hourly peak of 88 ppb with 7-hour averages ranging from 39 to 46 ppb. Seedlings in the 50 and 75% of ambient chambers were never exposed to greater than 80 ppb O(3). Visible foliar ozone injury (stipple) was significantly higher for R-12 seedlings than MO-7 seedlings and increased with increasing ozone exposures. For the chamber treatments averaged over all families, there was no significant difference in stomatal conductance and net photosynthetic rates, but there was a significant decrease in root biomass, and a significant decrease in root/shoot ratio between the 50 and 97% of ambient chambers. Stomatal conductance and net photosynthetic rates were significantly different between families with R-12 seedlings generally greater than MO-7 seedlings. The R-12 seedlings had a 7.5 mmol m(-2) increase in ozone uptake compared to MO-7, and at the same cumulative O(3) exposure R-12 exhibited 40.9% stippled leaf area, whereas MO-7 had 9.2% stippled leaf area. Significant differences were observed in stem volume growth and total final biomass between the open-top chambers and open plots. Although R-12 had the most severe foliar ozone injury, this family had significantly greater stem volume growth and total final biomass than MO-7 and NS seedlings. Root:shoot ratio was not significantly different between MO-7 and R-12 seedlings.

  Leaf morphology and ozone sensitivity of two open pollinated genotypes of black cherry (Prunus serotina) seedlings.:Environ Pollut. 2000 May;108(2):297-302.Ferdinand JA, Fredericksen TS, Kouterick KB, Skelly JM.The Environmental Resource Research Institute, The Pennsylvania State University, University Park, PA 16802, USA. jxf123@psu.edu

 Two black cherry families differing in sensitivity to ozone (O(3)) were used to test the hypothesis that leaf morphology is related to foliar sensitivity to O(3). Two uninjured sections of leaf tissue were sampled from a single leaf collected from each of 12 open-grown O(3)-tolerant and 12 O(3)-sensitive seedlings. Standard histological techniques and light microscopy were used to examine 11 morphological characteristics. The O(3)-sensitive genotype had significantly greater stomatal density, thinner palisade mesophyll layer and thicker spongy mesophyll layer, lower ratio of palisade to spongy mesophyll, greater leaf weight and leaf area. In addition, total leaf thickness, guard cell length, and specific leaf mass were also slightly greater for the O(3)-sensitive genotype.


  Development of negative feedback during successive growth cycles of black cherry.:Proc Biol Sci. 2004 Feb 7;271(1536):317-24.Packer A, Clay K.Department of Biology, Indiana University, Bloomington 47405, USA. packer@susqu.edu

 Negative feedback between plant and soil microbial communities can be a key determinant of vegetation structure and dynamics. Previous research has shown that negative feedback between black cherry (Prunus serotina) and soil pathogens is strongly distance dependent. Here, we investigate the temporal dynamics of negative feedback. To examine short-term changes, we planted successive cycles of seedlings in the same soil. We found that seedling mortality increased steadily with growth cycle when sterile background soil was inoculated with living field soil but not in controls inoculated with sterilized field soil. To examine long-term changes, we quantified negative feedback across successive growth cycles in soil inoculated with living field soil from a mature forest system (more than 70 years old) versus a younger successional site (ca. 25 years old). In both cases negative feedback developed similarly. Our results suggest that negative feedback can develop very quickly in forest systems, at the spatial scale of a single seedling.

  Physiological and foliar injury responses of Prunus serotina, Fraxinus americana, and Acer rubrum seedlings to varying soil moisture and ozone.:Environ Pollut. 2003;124(2):307-20.Schaub M, Skelly JM, Steiner KC, Davis DD, Pennypacker SP, Zhang J, Ferdinand JA, Savage JE, Stevenson RE.Graduate Ecology Program, Pennsylvania State University, University Park, PA 16802, USA.

 Sixteen black cherry (Prunus serotina, Ehrh.), 10 white ash (Fraxinus americana, L.) and 10 red maple (Acer rubrum, L.) 1-year old seedlings were planted per plot in 1997 on a former nursery bed within 12 open-top chambers and six open plots. Seedlings were exposed to three different ozone scenarios (ambient air: 100% O3; non-filtered air: 98% ambient O3; charcoal-filtered air: 50% ambient O3) within each of two different water regimes (nine plots irrigated, nine plots non-irrigated) during three growing seasons.During the 1998 and 1999 growing season, leaf gas exchange, plant water relations, and foliar injury were measured. Climatic data,ambient- and chamber-ozone-concentrations were monitored. We found that seedlings grown under irrigated conditions had similar (in 1998) but significantly higher gas exchange rates (in 1999) than seedlings grown within non-irrigated plots among similar ozone exposures. Cherry and ash had similar ozone uptake but cherry developed more ozone-induced injury (< 34% affected leaf area, LAA) than ash (<5% LAA), while maple rarely showed foliar injury, indicating the species differed in ozone sensitivity. Significantly more severe injury on seedlings grown under irrigated conditions than seedlings grown under non-irrigated conditions demonstrated that soil moisture altered seedling responses to ambient ozone exposures.

  Investigation of the microheterogeneity and aglycone specificity-conferring residues of black cherry prunasin hydrolases.:Plant Physiol. 2002 Jul;129(3):1252-64.Zhou J, Hartmann S, Shepherd BK, Poulton JE.Department of Biological Sciences, University of Iowa, Iowa City, Iowa 52242, USA.

 In black cherry (Prunus serotina Ehrh.) seed homogenates, (R)-amygdalin is degraded to HCN, benzaldehyde, and glucose by the sequential action of amygdalin hydrolase (AH), prunasin hydrolase (PH), and mandelonitrile lyase. Leaves are also highly cyanogenic because they possess (R)-prunasin, PH, and mandelonitrile lyase. Taking both enzymological and molecular approaches, we demonstrate here that black cherry PH is encoded by a putative multigene family of at least five members. Their respective cDNAs (designated Ph1, Ph2, Ph3, Ph4, and Ph5) predict isoforms that share 49% to 92% amino acid identity with members of glycoside hydrolase family 1, including their catalytic asparagine-glutamate-proline and isoleucine-threonine-glutamate-asparagine-glycine motifs. Furthermore, consistent with the vacuolar/protein body location and glycoprotein character of these hydrolases, their open reading frames predict N-terminal signal sequences and multiple potential N-glycosylation sites. Genomic sequences corresponding to the open reading frames of these PHs and of the previously isolated AH1 isoform are interrupted at identical positions by 12 introns. Earlier studies established that native AH and PH display strict specificities toward their respective glucosidic substrates. Such behavior was also shown by recombinant AH1, PH2, and PH4 proteins after expression in Pichia pastoris. Three amino acid moieties that may play a role in conferring such aglycone specificities were predicted by structural modeling and comparative sequence analysis and tested by introducing single and multiple mutations into isoform AH1 by site-directed mutagenesis. The double mutant AH ID (Y200I and G394D) hydrolyzed prunasin at approximately 150% of the rate of amygdalin hydrolysis, whereas the other mutations failed to engender PH activity.

  Hardwood seeding root and nutrient parameters for a model of nutrient uptake.:J Environ Qual. 2001 Mar-Apr;30(2):427-39.Kelly JM, Scarbrough JD, Mays PA.Dep. of Forestry, Iowa State Univ., Ames, IA 50011-1021, USA. jmkelly@iastate.edu

 Use of mechanistic models is an increasingly accepted way to evaluate complex processes. The Barber-Cushman model provides a means to simulate nutrient uptake once information on root system characteristics, nutrient uptake, and soil nutrient supply are developed. Objectives of this study were to determine during a growing season: (i) root growth for 1-yr-old black cherry (Prunus serotina Ehrh.), northern red oak (Quercus rubra L.), and red maple (Acer rubrum L.) seedlings; (ii) net plant increase in N, P, K, Ca, and Mg; (iii) soil solution and solid phase nutrient concentrations; and (iv) the influence of root growth and soil nutrient supply changes on nutrient uptake using the Barber-Cushman model. Seedlings were grown in pots containing A horizon soil from two forest sites. Measurements were made on five occasions during the growing season. Root growth averaged 41.5 cm d-1 for red maple compared with 28.0 and 16.7 cm d-1 for cherry and oak, respectively. Seventy-five percent of root growth occurred at the end of the growing season. Total plant N showed the greatest change (25-58%) due to soil source. Model simulations underestimated observed uptake by 31 to 99%. A clear relationship between soil solution nutrient concentration and plant uptake, an important assumption of the model, was not observed. Results indicate care will need to be exercised in the development and use of root growth and nutrient supply values in mechanistic models.

  Molecular analysis of (R)-(+)-mandelonitrile lyase microheterogeneity in black cherry.:Plant Physiol. 1999 Apr;119(4):1535-46.Hu Z, Poulton JE.Department of Biological Sciences, The University of Iowa, Iowa City, Iowa 52242, USA.

 The flavoprotein (R)-(+)-mandelonitrile lyase (MDL; EC 4.1.2.10), which plays a key role in cyanogenesis in rosaceous stone fruits, occurs in black cherry (Prunus serotina Ehrh.) homogenates as several closely related isoforms. Biochemical and molecular biological methods were used to investigate MDL microheterogeneity and function in this species. Three novel MDL cDNAs of high sequence identity (designated MDL2, MDL4, and MDL5) were isolated. Like MDL1 and MDL3 cDNAs (Z. Hu, J.E. Poulton [1997] Plant Physiol 115: 1359-1369), they had open reading frames that predicted a flavin adenine dinucleotide-binding site, multiple N-glycosylation sites, and an N-terminal signal sequence. The N terminus of an MDL isoform purified from seedlings matched the derived amino acid sequence of the MDL4 cDNA. Genomic sequences corresponding to the MDL1, MDL2, and MDL4 cDNAs were obtained by polymerase chain reaction amplification of genomic DNA. Like the previously reported mdl3 gene, these genes are interrupted at identical positions by three short, conserved introns. Given their overall similarity, we conclude that the genes mdl1, mdl2, mdl3, mdl4, and mdl5 are derived from a common ancestral gene and constitute members of a gene family. Genomic Southern-blot analysis showed that this family has approximately eight members. Northern-blot analysis using gene-specific probes revealed differential expression of the genes mdl1, mdl2, mdl3, mdl4, and mdl5.


  Sequencing, genomic organization, and preliminary promoter analysis of a black cherry (R)-(+)-mandelonitrile lyase gene.:Plant Physiol. 1997 Dec;115(4):1359-69.Hu Z, Poulton JE.Department of Biological Sciences, University of Iowa, Iowa City 52242, USA

 The flavoprotein (R)-(+)-mandelonitrile lyase (MDL; EC 4.1.2.10) plays a key role in cyanogenesis in rosaceous stone fruits. An MDL gene (mdl3) and its corresponding cDNA (MDL3) were isolated from black cherry (Prunus serotina) and characterized. The mdl3 gene contains 2292 bp of the 5' flanking region, the entire coding region, and 300 bp of the 3' flanking region. The coding region is interrupted by three short introns, of which one possesses the usual GC-AG splice junction dinucleotides. This gene encodes a polypeptide of 573 amino acids that includes a putative signal sequence, 13 potential N-glycosylation sites, and a presumptive flavin adenine dinucleotide-binding site. To determine whether the 5' flanking region of the mdl3 gene is capable of driving MDL expression, it was fused to the beta-glucuronidase reporter gene for Agrobacterium-mediated transformation into tobacco. Matching endogenous MDL expression patterns, beta-glucuronidase staining was observed in maturing embryos and seeds; it also occurred in postembryonic tissues, especially in association with vascular tissues. After developing a homologous transient transformation system to facilitate identification of putative regulatory sequences, we demonstrated that 125 bp (-107 to +18) of the 5' flanking sequence of the mdl3 gene is sufficient for MDL expression in protoplasts derived from immature black cherry embryos.

  Molecular cloning of acetone cyanohydrin lyase from flax (Linum usitatissimum). Definition of a novel class of hydroxynitrile lyases.:J Biol Chem. 1997 Feb 21;272(8):4770-4. Trummler K, Wajant H.Institute of Cell Biology and Immunology, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany.

 Acetone cyanohydrin lyase from Linum usitatissimum is a hydroxynitrile lyase (HNL) which is involved in the catabolism of cyanogenic glycosides in young seedlings of flax. We have isolated a full-length cDNA clone encoding L. usitatissimum HNL (LuHNL) from a cDNA expression library by immunoscreening. LuHNL cDNA was expressed in Escherichia coli and isolated from the respective soluble fraction in an active form which was biochemically indistinguishable from the natural enzyme. An open reading frame of 1266 base pairs encodes for a protein of 45,780 kDa. The derived amino acid sequence shows no overall homologies to the to date cloned HNLs, but has significant similarities to members of the alcohol dehydrogenase (ADH) family of enzymes. In particular, the cysteine and histidine residues responsible for coordination of an active site Zn2+ and a second structurally important Zn2+ in alcohol dehydrogenases are conserved. Nevertheless, we found neither alcohol dehydrogenase activity in LuHNL nor HNL activity in ADH. Moreover, well known inhibitors of ADHs, which interfere with the coordination of the active site Zn2+, fail to affect HNL activity of LuHNL, suggesting principally different mechanisms of cyanohydrin cleavage and alcohol oxidation. Interestingly, LuHNL like ADH and Prunus serotina (PsHNL) possesses an ADP-binding betaalphabeta unit motif, pointing to the possibility that the non-flavoprotein PsHNL and the flavoprotein LuHNL have developed from two independent lines of evolution of a common ancestor with an ADP-binding betaalphabeta unit.

  Evaluation of ozone injury on foliage of black cherry (Prunus serotina) and tall milkweed (Asclepias exaltata) in Great Smoky Mountains National Park.:Environ Pollut. 1997;95(1):13-8. Chappelka A, Renfro J, Somers G, Nash B.School of Forestry, Auburn University, Auburn, AL 36849, USA.

 The incidence and severity of visible foliar ozone injury on black cherry (Prunus serotina) seedlings and saplings and tall milkweed (Asclepias exaltata) plants in Great Smoky Mountains National Park (GRSM) were determined by surveys along selected trails conducted during late summer 1992. The incidence (% injured plants) of ozone injury on black cherry was 47% and the percent injured leaves/injured plant and average leaf area injured were 43 and 6%, respectively. Maximum severity (avg. leaf area of the most severely injured leaf) was 12%. Black cherry seedlings and saplings exhibiting ozone injury were taller than non-injured plants. When insect feeding was present, it occurred 96% of the time on plants with ozone injury. Significantly more injury (p=0.007) on black cherry (% injured leaves/injured black cherry) occurred in the NW section of GRSM compared with the other Park sections. Regression analyses showed no relationships in ozone injury with respect to aspect, slope or elevation. Tall milkweed was evaluated twice during August for ozone injury. The incidence (% injured plants) of ozone injury was 74 and 79% for the first and second survey, respectively. The percentage of injured leaves per plant from the first to second survey was 63 to 79%, respectively. Tall milkweeds showing ozone injury were taller than the non-injured plants. The percentage of insect-damaged plants was 50% among plants without ozone injury and 60% among ozone-injured plants. Non-injured tall milkweed had fewer flowers and/or pods than the injured plants. Mean leaf area injured increased over time, and mean maximum leaf area injured increased from 8 to 11% during the same period. Regression analyses showed no differences in ozone injury regarding aspect, slope or elevation. Our findings indicate that ozone injury is widespread throughout the Park on sensitive vegetation.

  Observations of ozone-induced foliar injury on black cherry (Prunus serotina, var. capuli) within the Desierto de Los Leones National Park, Mexico City.:Environ Pollut. 1997;95(2):155-8.Skelly JM, Savage JE, de Bauer Mde L, Alvarado D.Department of Plant Pathology, Pennsylvania State University, University Park, PA 16802, USA.

 A survey for ozone-induced foliar injury of black cherry was conducted in mid-June 1995 within the Desierto de Los Leones National Park located southwest of Mexico City. Evaluations of the upper and lower tree crowns of 18 trees revealed evidence of significant upper surface stipple, leaf reddening and premature senescence on 72% of the trees. A general survey of an additional 169 trees disclosed that 41% exhibited similar symptoms. A gradient of increasing symptoms with increasing elevation was also evident. For the most part, asymptomatic trees were observed to be situated within well-shaded coves at the lower elevations with very few symptomatic trees present in these areas.

  Light environment alters ozone uptake per net photosynthetic rate in black cherry trees.:Tree Physiol. 1996 May;16(5):485-90.Fredericksen TS, Kolb TE, Skelly JM, Steiner KC, Joyce BJ, Savage JE.Environmental Resources Research Institute, Pennsylvania State University, University Park, PA 16802, USA.

 Foliar ozone uptake rates of different-sized black cherry (Prunus serotina Ehrh.) trees were compared within a deciduous forest and adjacent openings in north-central Pennsylvania during one growing season. Study trees included open-grown seedlings and saplings, forest understory seedlings and saplings, and sunlit and shaded portions of mature canopy tree crowns. Instantaneous ozone uptake rates were highest in high-light environments primarily because of higher stomatal conductances. Low ozone uptake rates of seedlings and saplings in the forest understory could be attributed partially to lower average ambient ozone concentrations compared to the canopy and open environments. Among the tree size and light combinations tested, ozone uptake rates were highest in open-grown seedlings and lowest in forest-grown seedlings. Despite lower ozone uptake rates of foliage in shaded environments, ozone uptake per net photosynthesis of foliage in shaded environments was significantly higher than that of foliage in sunlit environments because of weaker coupling between net photosynthesis and stomatal conductance in shaded environments. The potential for greater ozone injury in shaded environments as a result of greater ozone uptake per net photosynthesis is consistent with previous reports of greater ozone injury in shaded foliage than in sunlit foliage.


  Temporal and spatial expression of amygdalin hydrolase and (R)-(+)-mandelonitrile lyase in black cherry seeds.:Plant Physiol. 1995 Sep;109(1):31-9.Zheng L, Poulton JE.Department of Biological Sciences, University of Iowa, Iowa City 52242, USA.

 In black cherry (Prunus serotina Ehrh.) macerates, the cyanogenic diglucoside (R)-amygdalin undergoes stepwise degradation to HCN catalyzed by amygdalin hydrolase (AH), prunasin hydrolase, and (R)-(+)-mandelonitrile lyase (MDL). A near full-length AH cDNA clone (pAH1), whose insert encodes the isozyme AH I, has been isolated and sequenced. AH I exhibits several features characteristic of beta-glucosidases of the BGA family, including their likely nucleophile center (isoleucine-threonine-glutamic acid-asparagine-glycine) and acid catalyst (asparagine-glutamic acid-proline/isoleucine) motifs. The temporal expression of AH and MDL in ripening fruit was analyzed by northern blotting. Neither mRNA was detectable until approximately 40 days after flowering (DAF), when embryos first became visible to the naked eye. Both mRNAs peaked at approximately 49 DAF before declining to negligible levels when the fruit matured (82 DAF). Taken together with enzyme activity data, these time courses suggest that AH and MDL expression may be under transcriptional control during fruit maturation. In situ hybridization analysis indicated that AH transcripts are restricted to the procambium, whereas MDL transcripts are localized within cotyledonary parenchyma cells. These tissue-specific distributions are consistent with the major locations of AH and MDL protein in mature seeds previously determined by immunocytochemistry (E. Swain, C.P. Li, and J.E. Poulton [1992] Plant Physiol 100:291-300).

  Physiology, morphology, and ozone uptake of leaves of black cherry seedlings, saplings, and canopy trees.:Environ Pollut. 1995;89(3):273-83.Fredericksen TS, Joyce BJ, Skelly JM, Steiner KC, Kolb TE, Kouterick KB, Savage JE, Snyder KR.Environmental Resources Research Institute, 220 Forest Resource Laboratory, Pennsylvania State University, University Park, Pennsylvania 16802, USA.

 Patterns of ozone uptake were related to physiological, morphological, and phenological characteristics of different-sized black cherry trees (Prunus serotina Ehrh.) at a site in central Pennsylvania. Calculated ozone uptake differed among open-grown seedlings, forest gap saplings, and canopy trees and between leaves in the upper and lower crown of saplings and canopy trees. On an instantaneous basis, seedling leaves had the greatest ozone uptake rates of all tree size classes due to greater stomatal conductance and higher concentrations of ozone in their local environment. A pattern of higher stomatal conductance of seedlings was consistent with higher incident photosynthetically-active radiation, stomatal density, and predawn xylem water potentials for seedlings relative to larger trees. However, seedlings displayed an indeterminate pattern of shoot growth, with the majority of their leaves produced after shoot growth had ceased for canopy and sapling trees. Full leaf expansion occurred by mid-June for sapling and canopy trees. Because many of their leaves were exposed to ozone for only part of the growing season, seedlings had a lower relative exposure over the course of the growing season, and subsequently lower cumulative uptake, of ozone than canopy trees and a level of uptake similar to upper canopy leaves of saplings. Visible injury symptoms were not always correlated with patterns in ozone uptake. Visible symptoms were more apparent on seedling leaves in concurrence with their high instantaneous uptake rates. However, visible injury was more prevalent on leaves in the lower versus upper crown of canopy trees and saplings, even though lower crown leaves had less ozone uptake. Lower crown leaves may be more sensitive to ozone per unit uptake than upper crown leaves because of their morphology. In addition, the lower net carbon uptake of lower crown leaves may limit repair and anti-oxidant defense processes.

  Immunocytochemical Localization of Prunasin Hydrolase and Mandelonitrile Lyase in Stems and Leaves of Prunus serotina.:Plant Physiol. 1994 Dec;106(4):1285-1291.Swain E, Poulton JE.Department of Biological Sciences, The University of Iowa, Iowa City, Iowa 52242.

 In macerates of black cherry (Prunus serotina Ehrh.) leaves and stems, (R)-prunasin is catabolized to HCN, benzaldehyde, and D-glucose by the sequential action of prunasin hydrolase (EC 3.2.1.21) and (R)-(+)-mandelonitrile lyase (EC 4.1.2.10). Immuno-cytochemical techniques have shown that within these organs prunasin hydrolase occurs within the vacuoles of phloem parenchyma cells. In arborescent leaves, mandelonitrile lyase was also located in phloem parenchyma vacuoles, but comparison of serial sections revealed that these two degradative enzymes are usually localized within different cells.

  Utilization of Amygdalin during Seedling Development of Prunus serotina.:Plant Physiol. 1994 Oct;106(2):437-445.Swain E, Poulton JE.Department of Biological Sciences, The University of Iowa, Iowa City, Iowa 52242.

 Cotyledons of mature black cherry (Prunus serotina Ehrh.) seeds contain the cyanogenic diglucoside (R)-amygdalin. The levels of amygdalin, its corresponding monoglucoside (R)-prunasin, and the enzymes that metabolize these cyanoglycosides were measured during the course of seedling development. During the first 3 weeks following imbibition, cotyledonary amygdalin levels declined by more than 80%, but free hydrogen cyanide was not released to the atmosphere. Concomitantly, prunasin, which was not present in mature, ungerminated seeds, accumulated in the seedling epicotyls, hypocotyls, and cotyledons to levels approaching 4 [mu]mol per seedling. Whether this prunasin resulted from amygdalin hydrolysis remains unclear, however, because these organs also possess UDPG:mandelonitrile glucosyltransferase, which catalyzes de novo prunasin biosynthesis. The reduction in amygdalin levels was paralleled by declines in the levels of amygdalin hydrolase (AH), prunasin hydrolase (PH), mandelonitrile lyase (MDL), and [beta]-cyanoalanine synthase. At all stages of seedling development, AH and PH were localized by immunocytochemistry within the vascular tissues. In contrast, MDL occurred mostly in the cotyledonary parenchyma cells but was also present in the vascular tissues. Soon after imbibition, AH, PH, and MDL were found within protein bodies but were later detected in vacuoles derived from these organelles.

  Tissue Level Compartmentation of (R)-Amygdalin and Amygdalin Hydrolase Prevents Large-Scale Cyanogenesis in Undamaged Prunus Seeds.:Plant Physiol. 1994 Jan;104(1):29-35.Poulton JE, Li CP.Department of Biological Sciences, The University of Iowa, Iowa City, Iowa 52242.

 Plum (Prunus domestica) seeds, which contain the cyanogenic diglucoside (R)-amygdalin and lesser amounts of the corresponding monoglucoside (R)-prunasin, release the respiratory toxin HCN upon tissue disruption. Amygdalin hydrolase (AH) and prunasin hydrolase (PH), two specific [beta]-glucosidases responsible for hydrolysis of these glucosides, were purified to near homogeneity by concanavalin A-Sepharose 4B and carboxymethyl-cellulose chromatography. Both proteins appear as polypeptides with molecular masses of 60 kD upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but they exhibit different isoelectric points (PH, 5.6-6.0; AH, 7.8-8.2). AH and PH were localized within mature plum seeds by tissue printing, histochemistry, and silver-enhanced immunogold labeling. As was previously shown in black cherry (Prunus serotina) seeds (E.Swain, C.P. Li, J.E. Poulton [1992] Plant Physiol 100: 291-300), AH and PH are restricted to protein bodies of specific procambial cells and are absent from the cotyledonary parenchyma, bundle sheath, and endosperm cells. In contrast, the cyanogenic glycosides in both plum and black cherry seeds, which were detected by tissue printing, occur solely in the cotyledonary parenchyma and are absent from the procambium and endosperm. It is concluded that tissue level compartmentation prevents large-scale cyanoglycoside hydrolysis in intact Prunus seeds.


  Leaf area and net photosynthesis during development of Prunus serotina seedlings.:Tree Physiol. 1993 Jan;12(1):55-69.Horsley SB, Gottschalk KW.USDA Forest Service, Northeastern Forest Experiment Station, Warren, PA 16365, USA.

 We used the plastochron index to study the relationship between plant age, leaf age and development, and net photosynthesis of black cherry (Prunus serotina Ehrh.) seedlings. Leaf area and net photosynthesis were measured on all leaves >/= 75 mm of plants ranging in age from 7 to 20 plastochrons. Effects of plant developmental stage on leaf area and net photosynthesis were evaluated for leaves of differing age (horizontal series), leaves on plants of constant age (vertical series), and leaves of constant age (oblique series). Regression techniques were used to estimate leaf area from leaf blade dimensions. The best equations for predicting leaf area had R(2) values of 0.991-0.992 and used linear or logarithmic functions of both leaf length and width. Suitable, but less precise, equations with R(2) values of 0.946-0.962 were developed from either leaf length or leaf width. Leaf area development in black cherry seedlings was similar to that in other indeterminate species. Leaves of young plants reached full expansion at a lower leaf plastochron age than leaves of older plants. Maximum net photosynthesis per unit leaf area occurred 2-3 plastochrons before full leaf expansion. There was strong ontogenetic drift in net photosynthesis with leaf age; net photosynthesis decreased as plant age increased in leaves of the same plastochron age. Plots of the oblique series were particularly useful in providing information about interaction effects.

  Tissue and Subcellular Localization of Enzymes Catabolizing (R)-Amygdalin in Mature Prunus serotina Seeds.:Plant Physiol. 1992 Sep;100(1):291-300.Swain E, Li CP, Poulton JE.Department of Biological Sciences, University of Iowa, Iowa City, Iowa 52242.

 In black cherry (Prunus serotina Ehrh.) homogenates, (R)-amygdalin is catabolized to HCN, benzaldehyde, and d-glucose by the sequential action of amygdalin hydrolase, prunasin hydrolase, and mandelonitrile lyase. The tissue and subcellular localizations of these enzymes were determined within intact black cherry seeds by direct enzyme analysis, immunoblotting, and colloidal gold immunocytochemical techniques. Taken together, these procedures showed that the two beta-glucosidases are restricted to protein bodies of the procambium, which ramifies throughout the cotyledons. Although amygdalin hydrolase occurred within the majority of procambial cells, prunasin hydrolase was confined to the peripheral layers of this meristematic tissue. Highest levels of mandelonitrile lyase were observed in the protein bodies of the cotyledonary parenchyma cells, with lesser amounts in the procambial cell protein bodies. The residual endosperm tissue had insignificant levels of amygdalin hydrolase, prunasin hydrolase, and mandelonitrile lyase.

  Prunus serotina Amygdalin Hydrolase and Prunasin Hydrolase : Purification, N-Terminal Sequencing, and Antibody Production.:Plant Physiol. 1992 Sep;100(1):282-290.Li CP, Swain E, Poulton JE.Department of Biological Sciences, University of Iowa, Iowa City, Iowa 52242.

 In black cherry (Prunus serotina Ehrh.) seed homogenates, amygdalin hydrolase (AH) participates with prunasin hydrolase (PH) and mandelonitrile lyase in the sequential degradation of (R)-amygdalin to HCN, benzaldehyde, and glucose. Four isozymes of AH (designated AH I, I', II, II') were purified from mature cherry seeds by concanavalin A-Sepharose 4B chromatography, ion-exchange chromatography, and chromatofocusing. All isozymes were monomeric glycoproteins with native molecular masses of 52 kD. They showed similar kinetic properties (pH optima, K(m), V(max)) but differed in their isoelectric points and N-terminal amino acid sequences. Analytical isoelectric focusing revealed the presence of subisozymes of each isozyme. The relative abundance of these isozymes and/or subisozymes varied from seed to seed. Three isozymes of PH (designated PH I, IIa, and IIb) were purified to apparent homogeneity by affinity, ion-exchange, and hydroxyapatite chromatography and by nondenaturing polyacrylamide gel electrophoresis. PH I and PH IIb are 68-kD monomeric glycoproteins, whereas PH IIa is dimeric (140 kD). The N-terminal sequences of all PH and AH isozymes showed considerable similarity. Polyclonal antisera raised in rabbits against deglycosylated AH I or a mixture of the three deglycosylated PH isozymes were not monospecific as judged by immunoblotting analysis, but also cross-reacted with the opposing glucosidase. Monospecific antisera deemed suitable for immunocytochemistry and screening of expression libraries were obtained by affinity chromatography. Each antiserum recognized all known isozymes of the specific glucosidase used as antigen.

  Ecophysiological and morphological responses to shade and drought in two contrasting ecotypes of Prunus serotina.:Tree Physiol. 1992 Jun;10(4):343-55.Abrams MD, Kloeppel BD, Kubiske ME.School of Forest Resources, The Pennsylvania State University, Ferguson Building, University Park, PA 16802, USA.

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

  Development of the Potential for Cyanogenesis in Maturing Black Cherry (Prunus serotina Ehrh.) Fruits.:Plant Physiol. 1992 Apr;98(4):1423-1428.Swain E, Li CP, Poulton JE.Department of Botany, University of Iowa, Iowa City, Iowa 52242.

 Biochemical changes related to cyanogenesis (hydrogen cyanide production) were monitored during maturation of black cherry (Prunus serotina Ehrh.) fruits. At weekly intervals from flowering until maturity, fruits (or selected parts thereof) were analyzed for (a) fresh and dry weights, (b) prunasin and amygdalin levels, and (c) levels of the catabolic enzymes amygdalin hydrolase, prunasin hydrolase, and mandelonitrile lyase. During phase I (0-28 days after flowering [DAF]), immature fruits accumulated prunasin (mean: 3 micromoles/fruit) but were acyanogenic because they lacked the above enzymes. Concomitant with cotyledon development during mid-phase II, the seeds began accumulating both amygdalin (mean: 3 micromoles/seed) and the catabolic enzymes and were highly cyanogenic upon tissue disruption. Meanwhile, prunasin levels rapidly declined and were negligible by maturity. During phases II (29-65 DAF) and III (66-81 DAF), the pericarp also accumulated amygdalin, whereas its prunasin content declined toward maturity. Lacking the catabolic enzymes, the pericarp remained acyanogenic throughout all developmental stages.


  Immunocytochemical Localization of Mandelonitrile Lyase in Mature Black Cherry (Prunus serotina Ehrh.) Seeds.:Plant Physiol. 1991 Aug;96(4):1329-1337.Wu HC, Poulton JE.Department of Botany, University of Iowa, Iowa City, Iowa 52242.

 Mandelonitrile lyase (MDL, EC 4.1.2.10), which catalyzes the reversible dissociation of (R)-(+)-mandelonitrile to benzaldehyde and hydrogen cyanide, was purified to apparent homogeneity from mature black cherry (Prunus serotina Ehrh.) seeds by conventional protein purification techniques. This flavoprotein is monomeric with a subunit molecular mass of 57 kilodaltons. Glycoprotein character was shown by its binding to the affinity matrix concanavalin A-Sepharose 4B with subsequent elution by alpha-methyl-d-glucoside. Upon chemical deglycosylation by trifluoromethanesulfonic acid, the molecular mass was reduced to 50.9 kilodaltons. Two-dimensional gel analysis of deglycosylated MDL revealed the presence of several subunit isoforms of similar molecular mass but differing slightly in isoelectric point. Polyclonal antibodies were raised in New Zealand white rabbits against deglycosylated and untreated MDL. Antibody titers were determined by enzyme linked immunosorbent and dot immunobinding assays, while their specificities were assessed by Western immunoblot analysis. Antibodies raised against untreated lyase recognized several proteins in addition to MDL. In contrast, antisera raised against deglycosylated MDL were monospecific and were utilized for developmental and immunocytochemical localization studies. SDS-PAGE and immunoblotting analysis of seed proteins during fruit maturation showed that MDL first appeared in seeds shortly after cotyledons began development. In cotyledon cells of mature seeds, MDL was localized primarily in the cell wall with lesser amounts in the protein bodies, whereas in endosperm cells, this labeling pattern was reversed. N-terminal sequence data was gathered for future molecular approaches to the question of MDL microheterogeneity.

  Black cherry growth response to ambient ozone and EDU.:Environ Pollut. 1991;70(3):241-54.Long RP, Davis DD.Department of Plant Pathology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.

 Field-grown black cherry (Prunus serotina Ehrh.) seedlings were treated with the antioxidant ethylenediurea (EDU) to evaluate height, diameter, and above-ground dry-weight biomass growth response to ambient ozone over four years. Nine blocks with 44 trees/block were used in a randomized complete block design with three foliar spray treatments: (1) 1000 ppm EDU mixed with a surfactant and water; (2) surfactant mixed with water; and (3) water only. In each growing season treatments were applied seven times at approximately 10-day intervals. Repeated measures analysis of variance indicated significant (P< or =0.05) treatment and year effects for log-transformed height and diameter growth over the four-year period. After four years, EDU-treated trees were approximately 17% taller and stem diameters were 21% greater than non-EDU-treated trees. Total above-ground dry-weight biomass at the end of four years was 47% greater for EDU-treated trees compared to non-EDU-treated trees.

  Localization and catabolism of cyanogenic glycosides.:Ciba Found Symp. 1988;140:67-91. Review.Poulton JE.Department of Botany, University of Iowa, Iowa City 52242.

 The catabolism of cyanogenic glycosides is initiated by cleavage of the carbohydrate moiety by one or more beta-glycosidases, which yields the corresponding alpha-hydroxynitrile. Until recently, the mode by which cyanogenic disaccharides are hydrolysed was largely unclear. Investigation of highly purified beta-glycosidases from plants containing cyanogenic disaccharides has now indicated that these compounds may be degraded via two distinct pathways, depending on the plant species. beta-Glycosidases from Davallia trichomanoides and Vicia angustifolia hydrolysed (R)-vicianin and (R)-amygdalin at the aglycone-disaccharide bond producing mandelonitrile and the corresponding disaccharide. Alternatively, hydrolysis of cyanogenic disaccharides in Prunus serotina, almonds, and Linum usitatissimum involves stepwise removal of the sugar residues. The nature of these enzymes and of other beta-glycosidases responsible for hydrolysis of simple cyanogenic monosaccharides is discussed. Hydroxynitriles may decompose either spontaneously or enzymically in the presence of a hydroxynitrile lyase to produce hydrogen cyanide and an aldehyde or ketone. The major kinetic and molecular properties of hydroxynitrile lyases purified from species accumulating aromatic and aliphatic cyanogens are reviewed. Cyanogenesis occurs rapidly only after cyanogenic plant tissues are macerated, allowing glycosides access to their catabolic enzymes. The possible nature of the compartmentation which prevents cyanogenesis under normal physiological conditions is discussed in relation to our knowledge of the tissue and subcellular localizations of cyanogens and catabolic enzymes.

  Isolation and characterization of multiple forms of prunasin hydrolase from black cherry (Prunus serotina Ehrh.) seeds.:Arch Biochem Biophys. 1987 May 15;255(1):19-26.Kuroki GW, Poulton JE.

 Three forms of prunasin hydrolase (PH I, PH IIa, and PH IIb), which catalyze the hydrolysis of (R)-prunasin to mandelonitrile and D-glucose, have been purified from homogenates of mature black cherry (Prunus serotina Ehrh.) seeds. Hydroxyapatite chromatography completely resolved PH I from PH IIa and PH IIb. PH IIa and IIb, which coeluted on hydroxyapatite, were resolved by gel filtration. PH IIa was a dimer with a native molecular weight of 140,000. Both PH I and PH IIb were monomeric with molecular weights of 68,000. The isozymes appeared to be glycoproteins based on their binding to concanavalin A-Sepharose 4B with subsequent elution by alpha-methyl-D-glucoside. When presented several potential glycosidic substrates, these enzymes exhibited a narrow specificity towards (R)-prunasin. Km values for (R)-prunasin for PH I, PH IIa, and PH IIb were 1.73, 2.3, and 1.35 mM, respectively. PH I and PH IIb possessed fivefold greater Vmax/Km values than PH IIa. Ortho- and para-nitrophenyl-beta-D-glucosides were hydrolyzed at the same active site. All forms had a pH optimum of 5.0 in citrate-phosphate buffer. PH I and PH IIb were competitively inhibited by castanospermine with Ki values of 0.19 and 0.09 mM, respectively. PH activity was not stimulated by any metal ion tested and was unaffected by diethyldithiocarbamate, o-phenanthroline, 2,2'-dipyridyl, and EDTA.

  Isolation and characterization of multiple forms of mandelonitrile lyase from mature black cherry (Prunus serotina Ehrh.) seeds.:Arch Biochem Biophys. 1986 Jun;247(2):440-5.Yemm RS, Poulton JE.

 Five multiple forms (forms 1-5) of mandelonitrile lyase (EC 4.1.2.10) which catalyze the decomposition of mandelonitrile to benzaldehyde and hydrogen cyanide have been extensively purified from seeds of black cherry (Prunus serotina Ehrh.) by concanavalin A-Sepharose 4B chromatography and chromatofocusing. These forms are monomers which differ only slightly in molecular weight (57,000-59,000) and isoelectric point (4.58-4.63), but heterogeneity in their carbohydrate side-chains was suggested by concanavalin A-Sepharose 4B chromatography. The absorption spectra of the predominating forms 4 and 5 showed maxima of 278, 380, and 460 nm, indicative of flavoprotein character. Detailed comparative kinetic studies of forms 4 and 5 revealed few significant differences in behavior. Both proteins showed pH optima between 6.0 and 7.0, had identical Km values (0.17 mM) for (R,S)-mandelonitrile, and retained similar activities upon storage at 4 and -20 degrees C. Neither form exhibited a metal ion requirement and both were affected similarly by metal salts, beta-mercaptoethanol, and sulfhydryl reagents. Benzoic acid, p-hydroxybenzyl alcohol, and benzyl alcohol inhibited both forms.


  Comparison of kinetic and molecular properties of two forms of amygdalin hydrolase from black cherry (Prunus serotina Ehrh.) seeds.:Arch Biochem Biophys. 1986 Jun;247(2):433-9.Kuroki GW, Poulton JE.

 Two forms of the beta-glucosidase amygdalin hydrolase (AH I and II), which catalyze the hydrolysis of (R)-amygdalin to (R)-prunasin and D-glucose, have been purified over 200-fold from mature black cherry (Prunus serotina Ehrh.) seeds. These proteins showed very similar molecular and kinetic properties but could be resolved by chromatofocusing and isoelectric focusing. AH I and II were monomeric (Mr 60,000) and had isoelectric points of 6.6 and 6.5, respectively. Their glycoprotein character was indicated by positive periodic acid-Schiff staining and by their binding to concanavalin A-Sepharose 4B with subsequent elution by alpha-Me-D-glucoside. Of the natural glycosidic substrates tested, both enzymes showed a pronounced preference for the endogenous cyanogenic disaccharide (R)-amygdalin. They also hydrolyzed at the same active site the synthetic substrates p-nitrophenyl-beta-D-glucoside and 4-methylumbelliferyl-beta-D-glucoside but were inactive towards (R)-prunasin, p-nitrophenyl-alpha-D-glucoside, and 4-methylumbelliferyl-alpha-D-glucoside. Maximum hydrolytic activity was shown in citrate-phosphate buffer in the pH range 4.5-5.0. AH I and II were inhibited competitively by the reaction product (R)-prunasin and noncompetitively (mixed type) by delta-gluconolactone and castanospermine.

  Xylem Development in Prunus Flower Buds and the Relationship to Deep Supercooling.:Plant Physiol. 1984 Apr;74(4):862-865.Ashworth EN.United States Department of Agriculture, Agricultural Research Service, Appalachian Fruit Research Station, Kearneysville, West Virginia 25430.

 Xylem development in eight Prunus species was examined and the relationship to deep supercooling assessed. Dormant buds of six species, P. armeniaca, P. avium, P. cerasus, P. persica, P. salicina, and P. sargentii deep supercooled. Xylem vessel elements were not observed within the dormant floral primordia of these species. Instead, discrete bundles containing procambial cells were observed. Vascular differentiation resumed and xylem continuity was established during the time that the capacity to deep supercool was lost. In P. serotina and P. virginiana, two species which do not supercool, xylem vessels ran the length of the inflorescence and presumably provided a conduit for the spread of ice into the bud. The results support the hypothesis that the lack of xylem continuity is an important feature of buds which deep supercool.

  The occurrence and seasonal distribution of C50-C60-polyprenols and of C100-and similar long-chain polyprenols in leaves of plants.:Acta Biochim Pol. 1984;31(1):115-26.Chojnacki T, Vogtman T.

 Large amounts of fully unsaturated, mainly-cis, higher isoprenoid alcohols consisting of 17-30 isoprene units were found in several plants of the Rosaceae family. They occur as mixtures of several prenologues with either C85 - or C100 - prenol dominating in the form of acetates. The highest level of these polyprenols (0.5-1.0% of wet weight) were found in Crataegus crus- galli , Cotonoaster lucida, Prunus serotina and Sorbus suecica (intermedia). Their content increased with increasing age of the leaves. The dynamics of this rise is different from that observed in the case of accumulation of free C50- C60 - prenols (up to 0.5% of wet weight) in leaves of various plant species.


  Phytosterol Pygeum africanum regulates prostate cancer in vitro and in vivo.:Endocrine. 2007 Feb;31(1):72-81.Shenouda NS, Sakla MS, Newton LG, Besch-Williford C, Greenberg NM, MacDonald RS, Lubahn DB.Department of Biochemistry, University of Missouri-Columbia, 920 East Campus Drive, 110A ASRC, Columbia, MO 65211, USA.

 BACKGROUND: Prostate cancer is an important public health problem. It is an excellent candidate disease for chemoprevention because prostate cancer is typically slow growing and is usually diagnosed in elderly males. Pygeum africanum (Prunus africana or Rosaceae) is an African prune (plum) tree found in tropical Africa. An extract from the bark of Pygeum africanum has been used in Europe as a prevention and treatment of prostate disorders including benign prostatic hypertrophy (BPH). More recently in the USA, the phytotherapeutic preparations of Pygeum africanum and Saw palmetto have been marketed for prostate health including prostate cancer prevention and treatment. METHODS: The anti-cancer potential of Pygeum africanum has been tested both in vitro (PC-3 and LNCaP cells) and in vivo (TRAMP mouse model). RESULTS: In tissue culture, ethanolic extracts (30%) of Pygeum africanum inhibited the growth of PC-3 and LNCaP cells; induced apoptosis and altered cell kinetics; down regulated ERalpha and PKC-alpha protein, and demonstrated good binding ability to both mouse uterine estrogen receptors and LNCaP human androgen receptors. TRAMP mice fed Pygeum africanum showed a significant reduction (P = 0.034) in prostate cancer incidence (35%) compared to casein fed mice (62.5%). CONCLUSION: Pygeum africanum, which is widely used in Europe and USA for treatment of BPH, has a significant role in regulation of prostate cancer both in vitro and in vivo and therefore may be a useful supplement for people at high risk for developing prostate cancer.

  Anti-proliferative effect of horehound leaf and wild cherry bark extracts on human colorectal cancer cells.:Oncol Rep. 2006 Jan;15(1):275-81.Yamaguchi K, Liggett JL, Kim NC, Baek SJ.Laboratory of Environmental Carcinogenesis, Department of Pathobiology, College of Veterinary Medicine, University of Tennessee, Knoxville, 37996, USA.

 Marubium vulgare (horehound) and Prunus serotina (wild cherry) have been traditionally used for the treatment of inflammatory-related symptoms such as cold, fever, and sore throat. In this report, we show that extracts of anti-inflammatory horehound leaves and wild cherry bark exhibit anti-proliferative activity in human colorectal cancer cells. Both horehound and wild cherry extracts cause suppression of cell growth as well as induction of apoptosis. We found that horehound extract up-regulates pro-apoptotic non-steroidal anti-inflammatory drug-activated gene (NAG-1) through transactivation of the NAG-1 promoter. In contrast, wild cherry extract decreased cyclin D1 expression and increased NAG-1 expression in HCT-116 and SW480 cell lines. Treatment with wild cherry extract resulted in the suppression of beta-catenin/T cell factor transcription, as assessed by TOP/FOP reporter constructs, suggesting that suppressed beta-catenin signaling by wild cherry extract leads to the reduction of cyclin D1 expression. Our data suggest the mechanisms by which these extracts suppress cell growth and induce apoptosis involve enhanced NAG-1 expression and/or down-regulation of beta-catenin signaling, followed by reduced cyclin D1 expression in human colorectal cancer cells. These findings may provide mechanisms for traditional anti-inflammatory products as cancer chemopreventive agents.

  The African cherry (Prunus africana): can lessons be learned from an over-exploited medicinal tree?.:J Ethnopharmacol. 2003 Nov;89(1):3-13. Review.Stewart KM.Keith and Schnars, P.A. 6500 North Andrews Avenue, Fort Lauderdale, FL, USA. StewartKristine@msn.com

 For the last 35 years, the African cherry (Prunus africana (Hook. f.) Kalm.) has been used in the treatment of benign prostatic hyperplasia and other disorders. The bark, from which the treatment is derived, is entirely wild-collected. The major exporters of bark include Cameroon, Madagascar, Equatorial Guinea, and Kenya. Groupe Fournier of France and Indena of Italy produce 86% of the world's bark extract, both for their own products and for the free market. Worldwide exports of dried bark in 2000 have been estimated at 1350-1525 metric tons per year, down from its peak of 3225 tons in 1997. Bark extracts (6370-7225 kg per year) are worth an estimated $4.36 million US dollars per year. In 2000, Plantecam, the largest bark exporter in Africa, closed its extraction factory in Cameroon, due to complex ecological, social, and economic factors. Wild-collection is no longer sustainable (and probably never was) where harvest seriously affects morbidity and mortality rates of harvested populations. Since 1995, it has been included in CITES Appendix II as an endangered species. In this paper, alternatives to wild-collection to meet future market demand are investigated, including conservation practices, enrichment plantings, small- and large-scale production, and protection of genetic resources. The species is at the beginning of a transition from an exclusively wild-collected species to that of a cultivated medicinal tree.

  Identification of components of Prunus africana extract that inhibit lipid peroxidation.:Phytomedicine. 1999 Nov;6(5):379-88.Hass MA, Nowak DM, Leonova E, Levin RM, Longhurst PA.Division of Basic and Pharmaceutical Sciences, Albany College of Pharmacy, NY 12208, USA. hassm@panther.acp.edu

 Extractive and chromatographic separations were performed on V-1326, a chloroform extract from the bark of Prunus africana (also referred to as Pygeum africanum), which is used to treat the symptoms associated with benign prostate hyperplasia (BPH). The relative amounts of eleven identified constituents in crude V-1326 and in separated fractions were determined using gas chromatographic analysis. The ability of V-1326 and its separated fractions to inhibit ferrous ion-induced stimulation of lipid peroxidation in microsomal preparations from rabbit livers was evaluated. The extract, V-1326, and fractions containing high levels of myristic acid potently inhibited lipid peroxidation.


  Scientific References:

  1.Research Update:Prunus serotina,Cherry Bark,Cherry Fruit.