Two new bisindole alkaloids from the seeds of Strychnos nux-vomica

The reinvestigation of Strychnos nux-vomica resulted in the isolation of two colorless monoquaternary bisindole alkaloids from the seeds. The structures of the two new compounds named 4-N-hydroxymethyl strychnidin-17-acetic acid (1) and 10, 11-dimethoxy-4-N-hydroxymethyl strychnidin-17-acetic acid (2), were defined by detailed spectral analyses, especially ¹H NMR, ¹³C NMR, DEPT, HSQC, ¹H NMR–¹H NMR, NOESY, HMBC and TOF-MS.     

Antidiarrhoeal activity of carbazole alkaloids from Murraya koenigii Spreng (Rutaceae) seeds

The bioassay guided fractionation of the n-hexane extract of the seeds of Murraya koenigii Spreng (Rutaceae) resulted in the isolation of three bioactive carbazole alkaloids, kurryam (I), koenimbine (II) and koenine (III). The structures of the compounds were confirmed from their ¹H-, ¹³C-, and 2D-NMR spectral data. Of the three compounds (I) and (II) exhibited significant inhibitory activity against castor oil-induced diarrhoea and PGE₂-induced enteropooling in rats. The compounds also produced a significant reduction in gastrointestinal motility in the charcoal meal test in Wister rats.     

A novel antifungal anthraquinone from seeds of Aegle marmelos Correa (family Rutaceae)

A new anthraquinone, 1-methyl-2-(3′-methyl-but-2′-enyloxy)-anthraquinone (1) has been isolated from seeds of Aegle marmelos Correa and was characterized on the basis of spectral analysis (UV, IR, ¹H NMR, ¹³C NMR, 2D NMR and mass spectroscopy). The compound exhibited significant antifungal activity against pathogenic strains of Aspergillus species and Candida albicans in disc diffusion assay (MIC value of 6.25 μg/disc), microbroth dilution and percent spore germination inhibition assays (MIC value of 31.25–62.5 μg/ml).     

Antioxidant and antibacterial activities of aqueous extract of Seabuckthorn (Hippophae rhamnoides) seeds

Seabuckthorn (Hippophae rhamnoides) seeds aqueous extract was screened for antioxidant and antibacterial activities. The antioxidant activities (reducing power, DPPH and liposome model system) showed a good antioxidant activity. The extract was also found to possess antibacterial activity with a MIC values with respect to Listeria monocytogenes and Yersinia enterocolitica found to be 750 and 1000 ppm, respectively. The antioxidant and antimicrobial effects of the extract implicate its potential for natural preservation.     

Study of anti-inflammatory,analgesic and antipyretic activities of seeds of Hyoscyamus niger and isolation of a new coumarinolignan

A chemical and biological validation of the traditional use of Hyoscyamus niger seeds as anti-inflammatory drug has been established. The methanolic extract of seeds of H. niger (MHN) was evaluated for its analgesic, anti-inflammatory and antipyretic activities in experimental animal models at different doses. MHN produced significant increase in hot plate reaction time, while decreasing writhing response in a dose-dependent manner indicating its analgesic activity. It was also effective in both acute and chronic inflammation evaluated through carrageenin-induced paw oedema and cotton pellet granuloma methods. In addition to its analgesic and anti-inflammatory activity, it also exhibited antipyretic activity in yeast-induced pyrexia model. Furthermore, the bioactive MHN under chemical investigation showed the presence of coumarinolignans as major chemical constituent and yielded a new coumarinolignan, cleomiscosin A methyl ether (1) along with four known coumarinolignans, cleomiscosin A (2), cleomiscosin B (3), cleomiscosin A-9′-acetate (4) and cleomiscosin B-9′-acetate (5). The structure elucidation of 1 was done by spectroscopic data interpretation and comparative HPLC analysis. Cleomiscosin A, but not its isomer cleomiscosin B, reduced dry and wet weight of cotton pellet granuloma in mice. This suggests that cleomiscosin A is an important constituent of MHN responsible for anti-inflammatory activity.     

Antioxidant and enzyme inhibition activities and chemical profiles of Polygonum sachalinensis F.Schmidt ex Maxim (Polygonaceae)

Polygonum sachalinensis is a widespread invasive plant in Europe. Chemical profiles of its different organs were studied by HPLC-UV-ESI/MS. Seven major constituents quercetin-3-O-β-D-galactopyranoside, quercetin-3-O-arabinopyranoside, lapathoside D, N-trans-feruloyltyramine, lapathoside C, hydropiperoside, and vanicoside B were isolated and identified. The free radical-scavenging, α/β-glucosidase, and acetylcholinesterase inhibitory activities of crude MeOH extracts and isolated compounds were studied. The structure–activity relationships were discussed. The chemical profiles revealed flavonoids and phenylpropanoids are the major compounds of all the organs of this plant. Quercetin-3-O-arabinopyranoside, lapathoside D, N-trans-feruloyltyramine, lapathoside C and hydropiperoside were isolated from this species for the first time. In the α-glucosidase bioassay, quercetin-3-O-β-D-galactopyranoside, lapathoside D and N-trans-feruloyltyramine demonstrated stronger activities than the positive reference acarbose. The trend in scavenging power showed no relation to enzyme inhibition in the test models.     

Adaptability of birch (Betula pendula Roth) and aspen (Populus tremula L.) genotypes to different soil moisture conditions

For northern Europe’s climate, models predict an increase in periods of drought and waterlogging. Knowledge of variation between genotypes of Betula pendula (birch) and Populus tremula (aspen) to drought and excess soil moisture are unavailable but relevant for future development of forest ecosystems. We studied variation between genotypes to soil moisture in birch and aspen with plant material representing naturally regenerated populations and showed differences in acclimation to soil moisture conditions. Genotypes showing most growth and biomass accumulation across treatments maintained most leaf area, high gas exchange and water-use efficiency and grew most root mass but had the lowest root length per unit root dry mass compared to other genotypes. This indicates that these genotypes are more efficient in harvesting water from the soil under adverse conditions. We also showed that birch and aspen employ different strategies to cope with soil moisture conditions, with aspen investing more in perennial parts, while birch efficiently maintains foliar processes. When the expression of some known drought responsive genes was measured, only ACC oxidase was induced by the drought treatment at the beginning of the experiment, while surprisingly LEA5, RD22 and ADH1 did not respond to drought, but were up-regulated in prolonged wet conditions indicating oxidative stress and hypoxia and that these genes are responding to multiple stress factors. We conclude that in plants micro-propagated from naturally regenerated birch and aspen populations, there is variation between genotypes in acclimation efficiency to soil moisture conditions.     

Annual variation in predation and dispersal of Arolla pine (Pinus cembra L.) seeds by Eurasian red squirrels and other seed-eaters

Seed predation and dispersal are key processes in the survival and distribution of plant species. Many animals cache seeds for later consumption, and, failing to recover some of these seeds, act as seed dispersers, influencing post-dispersal seed and seedling survival. Both animal and plant benefit from scatterhoarding and natural selection of seed characteristics and adaptations of seed predators (and dispersers) is one of the most important examples of co-evolution and mutualism. We studied the producer–consumer Arolla pine (Pinus cembra)–red squirrel (Sciurus vulgaris) system in a subalpine forest in the Italian Alps. Arolla pine produced large seed-crops (masting) at irregular intervals, followed by years with poor or moderate seed production. Squirrel density fluctuated in synchrony with the food resource, eliminating the time-lag normally present when resources are produced in pulses. In all years except 2009 (a mast-crop year), all Arolla pine cones were harvested (their seeds consumed and/or cached) by September to late October by different species. Both squirrels and nutcrackers (Nucifraga caryocatactes) fed on seeds, and their relative pre-dispersal predation rates (on cones in the canopy) differed between years. Overall, nutcrackers consumed more seeds between July and October than squirrels, but in 1 year squirrels took the largest number of seeds. Pre-dispersal seed predation by squirrels tended to be lower in years with large seed-crop size and there was a positive correlation, over the entire study period, between density of recovered hoards and Arolla pine seed density of the previous year. We conclude that (i) squirrels and nutcrackers are important pre-dispersal seed predators and seeds dispersers; (ii) squirrels are also post-dispersal seed predators, and (iii) the proportion of cached seeds consumed by squirrels increased with the size of the Arolla pine seed-crop, suggesting that red squirrel is a conditional mutualistic scatterhoarder of Arolla pine seeds.     

Anti-inflammatory, cyclooxygenase inhibitory and antioxidant activities of standardized extracts of Tridax procumbens L.

The standardized EtOAc, MeOH and 70% EtOH extracts of Tridax procumbens aerial parts showed significant inhibition of rat paw edema at a medium dose of 200 mg/kg and the EtOAC extract was the most active. These extracts were standardized by HPLC with the help of chemical markers. Further, the extracts were evaluated for COX-1 and COX-2 inhibitory activity and EtOAc extract exhibited the highest inhibition of COX-1 and COX-2 at 50 μg/mL. Cent aurein, centaureidin and bergenin were isolated as COX-1 and COX-2 inhibitory principles from the EtOAc extract. The extracts also exhibited antioxidant activity against DPPH and ABTS free radicals. The anti-inflammatory activity of T. procumbens aerial parts could be at least in part due to COX-1, COX-2 enzyme inhibition and free radical-scavenging activities which may be attributed to the presence of flavonoids and other polyphenols in the extracts.     

Nepetalactone content in shoot cultures of three endemic Nepeta species and the evaluation of their antimicrobial activity

Rapid micropropagation of Nepeta rtanjensis, N. sibirica and N. nervosa was performed. Qualitative and quantitative nepetalactone content in methanol extracts of in vitro grown plants was analysed by reverse-phase HPLC coupled with UV and MS detection. Only trans,cis-nepetalactone was detected in shoots of N. rtanjensis, while cis,trans-nepetalactone stereoisomer was present in N. sibirica. No nepetalactone was observed in shoots of N. nervosa. The antimicrobial activity of methanol extracts, against eight bacterial and eight fungal species, was evaluated. All the tested extracts showed significant antibacterial and strong antifungal activity. However, N. rtanjensis extract exhibited the best antimicrobial potential.     

Antinociceptive and anti-inflammatory activities of iridoid glycosides extract of Lamiophlomis rotata (Benth.) Kudo

Lamiophlomis rotata (Benth.) Kudo is a perennial herb (Labiatae) used as the Tibetan traditional medicine with the effects of alleviating pain, detumescence, hemostasis, promoting blood circulation to remove blood stasis and reinforcing marrow. In this study, we investigated the antinociceptive and anti-inflammatory activities of iridoid glycosides extract of L. rotata (IGLR) in mice. Our results showed that the iridoid glycosides extract could decrease acetic-acid-induced writhings times and formalin-induced lickings times, inhibit carrageenan-induced hind paw edema and xylene-induced ear swelling, and suppress peritoneal capillary permeability and leukocyte infiltration also induced by acetic acid in mice. All of these results suggested that the iridoid glycosides extract possesses the significant antinociceptive and anti-inflammatory activities.     

New amide alkaloid from the aerial part of Piper capense L.f. (Piperaceae)

Together with apigenine dimethylether and piperchabamide A, a new amide alkaloid, Kaousine and the Z form of antiepilepsirine were isolated from the aerial part of Piper capense L.f (Piperaceae). Their structures were elucidated by spectrometric methods and their in vitro antiparasitic activities were evaluated on Plasmodium falciparum.     

Biomass functions and expansion factors in young Norway spruce (Picea abies [L.] Karst) trees

Roots, stems, branches and needles of 160 Norway spruce trees younger than 10 years were sampled in seven forest stands in central Slovakia in order to establish their biomass functions (BFs) and biomass expansion factors (BEFs). We tested three models for each biomass pool based on the stem base diameter, tree height and the two parameters combined. BEF values decreased for all spruce components with increasing height and diameter, which was most evident in very young trees under 1 m in height. In older trees, the values of BEFs did tend to stabilise at the height of 3–4 m. We subsequently used the BEFs to calculate dry biomass of the stands based on average stem base diameter and tree height. Total stand biomass grew with increasing age of the stands from about 1.0 Mg ha⁻¹ at 1.5 years to 44.3 Mg ha⁻¹ at 9.5 years. The proportion of stem and branch biomass was found to increase with age, while that of needles was fairly constant and the proportion of root biomass did decrease as the stands grew older.     

Regeneration of Rocky Mountain bristlecone pine (Pinus aristata) and limber pine (Pinus flexilis) three decades after stand-replacing fires

Rocky Mountain bristlecone pine (Pinus aristata) and limber pine (Pinus flexilis) are important high-elevation pines of the southern Rockies that are forecast to decline due to the recent spread of white pine blister rust (Cronartium ribicola) into this region. Proactive management strategies to promote the evolution of rust resistance and maintain ecosystem function require an improved understanding of the role of disturbance on the population dynamics of both species and environmental conditions that favor seedling establishment. We examined patterns of bristlecone and limber pine regeneration across the perimeters of three, 29-year-old, high-severity burns in northern, central, and southern Colorado: Ouzel, Badger Mountain, and Maes Creek, respectively. Both species exhibited a very protracted regeneration response to these fires. Bristlecone pine regeneration was concentrated near burn edges and beneath surviving seed sources. This spatial pattern is consistent with limitations incurred by wind-dispersal, also borne out by the low occurrence of seedling clusters. Relative to unburned stands, the absolute abundance of bristlecone pine generally increased only on plots retaining some surviving trees. Limber pine regeneration pattern varied between sites: high in the burn interior at Ouzel, concentrated at burn edges at Badger, and mostly in unburned stands at Maes. Clark’s Nutcracker dispersal of limber pine in each study area was indicated by high seedling distance from possible seed sources and high frequencies of clustered stems. Except at Ouzel, the absolute abundance of limber pine decreased in burns. Across sites, establishment by both species was boosted by nearby nurse objects (rocks, fallen logs, and standing tree trunks), a relationship that extended out at least as far as the closest three such objects, usually found within 50 cm. Fire decreased the frequency of Pedicularis but increased Castilleja and Ribes species (alternate hosts of white pine blister rust), though only one species, R. cereum, was positively associated with either pine species. We conclude that regeneration of bristlecone and limber pine may benefit from natural disturbance or proactive management creating appropriately sized openings and microtopographic structure (e.g., abundant fallen logs); however, beneficial responses may require many decades to be achieved.     

日本落叶松咖啡酸-O-甲基转移酶基因 LkCOMT的克隆及单核苷酸多态性分析

依据日本落叶松转录组数据库检测到的咖啡酸-O-甲基转移酶(COMT)基因ESTs序列设计引物,分离得到一个编码该酶的新基因。其cDNA克隆全长为1 350 bp,包含长度为1 092 bp的开放阅读框,可编码364个氨基酸残基。序列分析显示,所推导的蛋白质氨基酸序列包含COMT基因5个所特有的保守元件,与海岸松PpCOMT的蛋白质氨基酸序列同源性达94%。在此基础上,利用DnaSP5.0软件对日本落叶松40株基因型个体的LkCOMT序列进行了单核苷酸多样性SNPs分析,共检测到92个SNP位点,SNP发生频率为1/17 bp,多样性指数π_T为0.007 80。在这些SNPs中,65个属于转换,27个属于颠换。在外显子区域,共检测到58个SNP位点,其中37个为错义突变,21个为同义突变。研究结果为日本落叶松基因标记辅助育种提供了理论基础。     

Growth and morphology of three forest understorey species (Calluna vulgaris, Molinia caerulea and Pteridium aquilinum) according to light availability

Calluna vulgaris, Molinia caerulea and Pteridium aquilinum are three forest understorey species that compete with tree seedlings for environmental resources. Forest managers therefore seek to control their growth, which is driven in part by the light available in the forest understorey.An experiment was set up in a nursery in which the three understorey species C. vulgaris, M. caerulea and P. aquilinum were planted at six light levels (6, 11, 22, 36, 48 and 100% of full sunlight) replicated in three blocks. The different light levels other than 100% were obtained using neutral shading nets. The response of the three species to light availability was estimated after two growing seasons by their cover and height, their morphology, assessed by the foliar tuft height-to-width ratio for C. vulgaris and M. caerulea and by the frond height-to-length ratio for P. aquilinum, and their functional acclimation, assessed by a leaf trait (leaf mass on an area basis, LMA).Vegetation cover changed with light availability following a bell-shaped curve. By contrast, vegetation height remained approximately the same, irrespective of the light treatment, except for P. aquilinum, which grew higher than the other two species at medium and high light levels. As a consequence, vegetation individuals were more spindly in a dark environment. From the lowest light treatment, P. aquilinum cover was high (75%) and intercepted a larger proportion of the light than the other two species. This species thus proved the most competitive for light.All three species showed a notable acclimation to light availability by their LMA, though in different ways. The LMA of C. vulgaris decreased in shaded conditions by an increase in leaf area, whereas decrease in LMA for P. aquilinum was due to a decrease in leaf mass. For M. caerulea, the LMA remained stable whatever the light conditions by a simultaneous increase in both leaf mass and leaf area in shaded conditions.Consequently, the in-forest development of C. vulgaris and M. caerulea could be managed through small adult canopy thinnings, as both species grew well from light levels above 20%. Conversely, it seems unrealistic to manage P. aquilinum in this way as its development was almost maximal even at very low light levels.     

The influence of essential oil of aniseed (Pimpinella anisum, L.) on drug effects on the central nervous system

Anise (Pimpinella anisum L.; Apiaceae) and its essential oil have been widely used in folk medicine, pharmacy and food industry. Since there are some data about the impact of anise on functions of central nervous system (CNS), the issue of possible interactions with drugs acting in CNS should be considered. This survey aimed to examine the influence of aniseed essential oil (EO) intake on the effects of drugs that act in CNS.     

The effects of clearcuts and forest buffer size on post-breeding emigration of adult wood frogs (Lithobates sylvaticus)

The establishment of terrestrial buffer zones around vernal pools has been recommended to provide upland habitat for pool-breeding amphibians in areas where forestry practices occur adjacent to breeding sites. However, few studies have empirically tested the effectiveness of buffers. We assessed post-breeding emigration behavior (net emigration distance, rate of movement, proportionate use of available habitats) of radio-tagged adult wood frogs (Lithobates sylvaticus) at nine vernal pools with experimental forest buffer treatments in central Maine, USA. Buffer treatments were either 30-m (N = 3) or 100-m (N = 3) forest buffers surrounded by a 100-m wide clearcut; pools surrounded by uncut forest served as reference sites (N = 3). We tracked 33 individuals in 2004 and 2005, for an average of 41 days, as they emigrated from breeding pools. Recently clearcut habitat was permeable to emigrating adult wood frogs, particularly females. A higher proportion of frogs at 30-m buffer sites than at 100-m buffer sites traveled through the clearcuts to reach intact forest beyond, suggesting that 30-m buffers may not provide sufficient upland habitat to support adult wood frog populations. There was high variability in emigration behavior among frogs, regardless of buffer treatment, and males and females tended to exhibit differential responses to different buffer sizes and to clearcut habitat. Although wood frogs in this study utilized both 30-m and 100-m forest buffers, variability between sexes and density-dependent effects could render small buffers (e.g., 30 m or less) inadequate to support these populations.     

松褐天牛肿腿蜂对松褐天牛低龄幼虫控制作用的研究

研究测定了松褐天牛肿腿蜂对松褐天牛1～3龄幼虫的控制作用,采用林间和室内罩网相结合的方法研究了2组松褐天牛肿腿蜂对松褐天牛幼虫的寄生情况.结果显示:在室内试验条件下,松褐天牛肿腿蜂产卵之前的刺蛰、补充营养取食能直接造成松褐天牛1～3龄幼虫死亡,对1、2及3龄幼虫的平均致死率分别为95.66％、85.91％、57.88％,并可产卵寄生2、3龄幼虫;林间释放2组松褐天牛肿腿蜂后,在刺蛰和补充营养取食后能有效寄生寄主的肿腿蜂雌蜂数量比例变化较大,有效肿腿蜂率最高为14.9％,最低为2.3％.松褐天牛肿腿蜂寄生行为不受寄主聚集程度的影响,但刚羽化出茧肿腿蜂雌蜂的寄生效率比经饲喂后的雌蜂高近5倍.从肿腿蜂单一个体来看,每头雌蜂补充营养取食和产卵寄生致死的松褐天牛幼虫数为2.1～7.7头.松褐天牛肿腿蜂雌蜂的补充营养习性及刺蛰、取食致死寄主行为在防治中有很重要的作用,表明该种肿腿蜂是防治松褐天牛的重要天敌,在生物防治松褐天牛幼龄幼虫上具有良好的应用前景.     

Masting behaviour and dendrochronology of European beech (Fagus sylvatica L.) in southern Sweden

To identify weather controls of beech diameter growth and masting in southern Sweden, we analyze records of monthly weather, regional masting record, and tree-ring chronologies from five beech-dominated stands. The results indicate a strong weather control of temporal pattern of masting events in southern Sweden over the second half of the 20th century. Negative summer temperature anomaly 2 years prior to a mast year, coupled with positive temperature anomaly in the year immediately preceding the same mast year, is a characteristic weather pattern associated with known mast years. Strong dependence of beech masting behaviour on temperature explains the high degree of regional synchronization of masting events. Growth of beech in southern Sweden is strongly and negatively correlated with previous year's summer temperature and positively – with previous year's October temperature. The present study does not provide a conclusive answer in identifying a full set of direct and indirect effects of climatic variables controlling tree-ring growth, since the negative effect of previous year's summer temperature may be a result of a temperature-controlled increase in the beech nut production in the current year. Consistent and significant negative departures of ring-width index during mast years support the hypothesis about a trade-off between investment of bioassimilates into production of beech nuts and tree-ring growth. Alternative explanation of growth anomalies in mast years, relating such anomaly to a negative impact of previous year's growing season, was not supported by the data. We found a limited effect of masting on diameter growth in the following years, indicating that decline in the overall wood production, associated with heavy masting, is short term and typically occurs in the year of actual masting.