Screening of Indonesian medicinal plants for inhibitor activity on nitric oxide production of RAW264.7 cells and antioxidant activity

Traditional Indonesian medicinal plants were screened for their inhibitory effects on the nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages and for the antioxidant activity through the evaluation of free radical scavenging effect and reducing power. The results of screening indicated that 50 methanolic extracts inhibited (>50%) lipopolysaccharides (LPS)-induced NO release from RAW264.7 cells at 50 microg/ml, with 18 having greater than 100% inhibition. At 200 microg/ml, 61 methanol extracts exhibited inhibitory activity (>50%), with 45 showing greater than 100% inhibition. In addition, the free radical scavenging effects of 6 methanolic extracts were found to be more than 50% for extract concentration of 0.5 mug/ml. The results indicate that the extracts contain active compounds that inhibit NO release and scavenge free radical.     

Antifungal properties of Pseudarthria viscida

The extracts of leaf, root, stem, and the callus obtained from Pseudarthria viscida showed significant inhibitory activity against some fungal pathogens causing major diseases in crop plants and stored food grains.     

Uptake and distribution of calcium and phosphorus in beech (Fagus sylvatica) as influenced by aluminum and nitrogen

We studied the effects of excess nitrogen added as nitrate (NO(3) (-)) or ammonium (NH(4) (+)), or both, on mineral nutrition and growth of beech (Fagus sylvatica L.) plants grown at pH 4.2 in Al-free nutrient solution or in solutions containing 0.1 or 1.0 mM AlCl(3). A high external concentration of NH(4) (+) increased the concentration of nitrogen in roots, stems and leaves. The root/shoot dry weight ratio was less in plants grown in the presence of NH(4) (+) than in plants grown in the presence of NO(3) (-). The concentration of phosphorus in the roots was increased and the concentration of potassium in all parts of the plant was decreased by NH(4) (+). A high external concentration of NO(3) (-) caused a decrease in phosphorus concentrations of the root, stem and leaf. Uptake of (45)Ca(2+) by roots was reduced in the presence of high concentrations of NH(4) (+) or NO(3) (-), and a combination of high concentrations of nitrogen and aluminum further reduced the uptake of (45)Ca(2+). Uptake of phosphate ((32)P) and concentrations of phosphorus in root and shoot were increased when plants were grown in the presence of 0.1 mM Al. Exposure to 1.0 mM Al, however, reduced the concentration of phosphorus in roots and shoots and the reduction was greater when plants were grown in the presence of a high external NO(3) (-) concentration. Aluminum binds to roots, and plants grown in the presence of 1.0 mM Al had a slightly higher concentration of aluminum in roots than plants grown in the presence of 0.1 mM Al, whereas the concentration of Al in the shoot was increased 2 to 3 times in plants exposed to 1.0 mm Al. Furthermore, the effects of 1.0 mM Al on uptake of other macronutrients were quite different from the effects of 0.1 mM Al. We conclude that 0.1 mM Al facilitates uptake and transport of phosphorus in beech and that between 0.1 and 1.0 mM Al there is a dramatic change in the effects of Al on uptake and transport of divalent cations and phosphorus.     

Role for toll-like receptor 4 in TNF-alpha secretion by murine macrophages in response to polysaccharide Krestin,a Trametes versicolor mushroom extract

Woody fungi and yeast preparations show promise in cancer treatment by activating anti-tumor immune responses. Macrophages (J774A.1) were treated with PSK, Reishi extract, scleroglucan or vehicle control. Pre-incubation with TLR4 blocking antibody inhibited TNF-alpha secretion by both J774A.1 cells and primary splenocytes but had inconclusive effect on scleroglucan-induced secretion of TNF-alpha. PSK induced TNF-alpha and IL-6 secretion by wild type but not by TLR4-deficient peritoneal macrophages. We conclude that constituents from PSK act as ligands for TLR4 receptors leading to induction of TNF-alpha and IL-6 inflammatory cytokines. Receptor-mediated differences may be due to structural differences in beta glucans or non-glucan constituents.     

Responses to ultraviolet-B radiation by purely symbiotic and NO3-fed nodulated tree and shrub legumes indigenous to southern Africa

Purely symbiotic and NO3-fed nodulated seedlings of Virgilia oroboides (Bergius) T.M. Salter, Cyclopia maculata (L.) Vent and Podalyria calyptrata Willd. were exposed to biologically effective ultraviolet-B radiation (UV-B) to assess the effects of above- and below-ambient UV-B on growth, symbiotic function and metabolite concentrations. Seedlings were grown outdoors either on tables under ambient or 34 or 66% above-ambient UV-B conditions (UV-B100 control, UV-B134 and UV-B166, respectively), or in chambers providing below-ambient (22% of ambient) UV-B (UV-B22) along with a UV-A control and a photosynthetically active radiation (PAR) control. Exposure of seedlings to UV-B166 radiation reduced (P < or = 0.05) leaf and stem dry mass by 34 and 39%, respectively, in C. maculata, and reduced leaf nitrogen concentration (%N) by 12% in V. oroboides. Nodule %N in C. maculata and stem %N in P. calyptrata also decreased (P < or = 0.05) in response to UV-B22 radiation compared with the UV-A control, but not compared with the PAR control. Concentrations of flavonoids, soluble sugars and starch were unaltered by the UV-B treatments. Application of 1 mM NO3 to UV-B166-treated seedlings increased whole-plant dry mass of V. oroboides and P. calyptrata by 47 and 52%, respectively. Dry mass of organs, nodule %N and total N concentration of these species also increased with NO3 application. However, NO3 supply decreased (P < or = 0.05) nodule dry mass, stem %N and leaf %N as well as root and leaf anthocyanin concentrations in C. maculata. In terms of UV-B x N interactions, dry mass of stems, roots, nodules and total biomass of NO3-fed C. maculata seedlings were reduced, and nodule %N, total N and leaf anthocyanins were depressed by the UV-B134 and UV-B166 treatments relative to UV-B100-treated seedlings. Although we found that above-ambient UV-B had no effects on growth and symbiotic function of V. oroboides and P. calyptrata seedlings, feeding NO3 to these species increased (P < or = 0.05) seedling growth. In contrast, purely symbiotic C. maculata seedlings were sensitive to the UV-B166 radiation treatment, and adding NO3 further increased their sensitivity to both the UV-B134 and UV-B166 treatments.     

大荨麻根、茎叶乙醇提取物化学成分的GC/MS研究

利用气相色谱-质谱联用技术对大荨麻根、茎叶乙醇提取物化学成分进行分析，结果表明，茎叶醇提物的19种化学成分主要以脂肪酸及其衍生物、硼烷配合物、杂环类化合物为主，其含量分别为51．05％、21．19％、13．18％，其中医药化工原料中间体残留物占52．51％；根醇提物的17种化学成分主要以脂肪酸及其衍生物、杂环类化合物、烃类化合物为主，分别含50．77％、30．71％、10．62％，其中农药残留物占29．68％。     

苍耳对苹果轮纹病菌等的抑菌活性研究

以苍耳根、茎、叶作为试验材料,以丙酮、80%乙醇、乙酸乙酯、蒸馏水作为提取溶剂,利用滤纸片法研究了苍耳不同器官4种不同溶剂提取液对2种细菌和1种真菌的抑菌活性,结果表明:苍耳各器官的不同溶剂提取液抑菌效果不同。对溶剂来说,苍耳叶丙酮、乙酸乙酯提取液对枯草芽孢杆菌的抑菌圈直径分别达16.32mm、15.66mm,叶的乙酸乙酯提取液对苹果轮纹病菌的抑菌圈直径达15.64mm,4种不同溶剂提取液的总体抑菌活性由大到小依次为丙酮、乙酸乙酯、80%乙醇、蒸馏水;对不同器官来说,苍耳各器官提取液抑菌效果由大到小依次为叶、根、茎,尤以叶的丙酮提取液抑菌效果最佳,乙酸乙酯次之。苍耳叶的丙酮、乙酸乙酯提取液对枯草芽孢杆菌、苹果轮纹病菌均表现为较强的抑菌活性,但对大肠杆菌的抑菌活性较弱。     

Hebeloma crustuliniforme facilitates ammonium and nitrate assimilation in trembling aspen (Populus tremuloides) seedlings

This study examined the role of ectomycorrhizal associations in nitrogen assimilation of Populus tremuloides seedlings. Seedlings were inoculated with Hebeloma crustuliniforme and compared with non-inoculated plants. Nitrogen-metabolizing enzymatic properties were also determined in H. crustuliniforme grown in sterile culture. The seedlings and fungal cultures were subjected to nitrogen treatments (including NO??, NH?? and a combination of NO???+?NH??) for 2 months to examine the effects on growth, nitrogen-assimilating enzyme activities and xylem sap concentrations of NH?? and NO??. Seedlings were also provided for 3 days with 1?N-labeled NH?? and NO??, and leaf and root 1?N content relative to total nitrogen was measured. Both NO?? and NH?? were effective in supporting seedling growth when either form was provided separately. When NO?? and NH?? were provided together, seedling growth decreased while enzymatic assimilation of NH?? increased. Additionally, nitrogen assimilation in inoculated seedlings was less affected by the form of nitrogen compared with non-inoculated plants. Fungal ability to enzymatically respond to and assimilate NH?? combined with aspen's enzymatic responsiveness to NO?? was likely the reason for efficient assimilation of both nitrogen forms by mycorrhizal plants.     

In vitro plant regeneration of Eclipta alba and increased production of coumestans

Eclipta alba plants were regenerated from totipotent calli of leaf, stem and root cultured on Murashige and Skoog’s medium supplemented with various growth regulators. The production of the coumestans wedelolactone and demethylwedelolactone was estimated by UV method.     

Observation of the scale of patchy stomatal behavior in leaves of Quercus crispula using an Imaging-PAM chlorophyll fluorometer

Patchy stomatal closure occurs in plants with heterobaric leaves, in which vertical extensions of bundle sheath cells delimit the mesophyll and restrict the diffusion of CO(2). The scale of patchy stomatal behavior was investigated in this study. The distribution of PSII quantum yield (Φ(II)) obtained from chlorophyll fluorescence images was used to evaluate the scale of stomatal patchiness and its relationship with leaf photosynthesis in the sun leaves of 2-year-old saplings of Quercus crispula Blume. Fluorescent patches were observed only during the day with low stomatal conductance. Comparison of numerical simulation of leaf gas exchange and chlorophyll fluorescence images showed that heterogeneous distribution of electron transport rate through PSII (J) was observed following stomatal closure with a bimodal manner under both natural and saturated photosynthetic photon flux densities. Thus, fluorescence patterns can be interpreted in terms of patchy stomatal closure. The mapping of J from chlorophyll fluorescence images showed that the scale of stomatal patchiness was approximately 2.5-fold larger than that of anatomical patches (lamina areas bounded by bundle sheath extensions within lamina). Our results suggest the spatial scale of stomatal patches in Q. crispula leaves.     

In vitro and in vivo immune stimulating effects of a new standardized Echinacea angustifolia root extract (Polinacea)

Polinacea is a new standardized hydroethanolic extract obtained from Echinacea angustifolia roots containing echinacoside (>4%), the high molecular weight polysaccharide IDN 5405 (>5%) and a isobutylamide fraction (<0.1%). For in vitro tests, a bacterial lipopolysaccharide-free (LPS-free) Polinacea has been prepared in order to avoid non-specific responses of immunocompetent cells. LPS-free Polinacea enhanced the immune functions as highlighted by the proliferation rate and gamma-interferon production in murine T-lymphocyte cell cultures stimulated by anti-CD3. LPS-free Polinacea did not have a direct role on macrophage response as measured in the nitric oxide production test using the J774 macrophage cells line. In vivo, Polinacea showed an immune stimulating activity by reducing the Candida albicans induced mortality both in normal and in cyclosporin A-treated mice.     

Leaf growth of Eucalyptus globulus seedlings under water deficit

Eucalyptus globulus Labill. seedlings grown under field conditions in Portugal were watered either daily (control) or every 6 days (drought-treated). Relative to those of control plants, rates of leaf production and leaf biomass accumulation were reduced by almost half in drought-treated plants. However, whereas expansion of new leaves on control plants slowed toward the end of the 30 day experiment, expansion of leaves of the same age on drought-treated plants accelerated as a change in weather conditions resulted in midday plant water potentials above -3.0 MPa. In plants that were left unwatered until they wilted and were then watered daily, expansion of the fifth leaf pair from the apex was slower than that of the same pair of leaves of plants watered daily throughout; but it continued for about twice as long and resulted in the same final leaf area. Drought treatment also caused a substantial reduction in the rate of leaf production, which, in part, accounted for the effect of drought on leaf biomass production. In a greenhouse study, witholding water for 15 days had only a slight effect on the length or width of adaxial epidermal cells, and the effect was quickly reversed on rewatering.     

根癌农杆菌对感染植原体的泡桐组培苗症状的影响

采用含有激素合成相关基因的根癌农杆菌，伤口接种已感染植原体的泡桐丛植组培苗和健康组培苗，结果发现对丛植苗的致瘤能力明显低于健康对照苗，且被接种病苗的丛枝症状缓解，从健苗获得的T-DNA转化泡桐瘤组织细胞能在无激素培养基上稳定生长和连续继代培养2年以上，说明瘤组织细胞自身已获得了细胞分裂素和生长素合成能力，根据已报道的根癌农杆菌株系pTil5955T-DNA的异戊烯基转移酶基因（ipt)的保守序列，设计了一对引物（CYT和CYT′），用多聚酶链式反应（PCR）扩增了我国杨树致瘤农杆菌ipt基因部分序列（427bp片段），也从遗传转化的两个泡桐无性系瘤组织At-ZH和At-T35扩增出此特异片估，从而进一步肯定了T-DNA已被整合到泡桐的染色体上表明泡桐易于通过Ti质粒载体途径进行基因转移操作，但用此引物未能从泡桐、甘薯健株和感染植原体的组培病苗扩增出相应的427bp特异片段，当用此遗传转化瘤组织嫁接病苗时，可减轻从枝症状的严重度，延长病苗的存活时间和诱导病株生根，这进一步证实了泡桐在与植原体相互作用过程中激素代谢发生了变化。     

Effects of various stress treatments on growth and ethylene evolution in seedlings and sprouts of betula pendula Roth and B. pubescens Ehrh.

Experiments were conducted to determine growth and ethylene evolution in seedlings and coppiced plants of Betula pendula Roth and B. pubescens Ehrh. when subjected to stresses relevant to the maintenance of natural forests, and especially in fuelwood plantations, i.e. cutting of the stem, thinning, bending, flooding and various combinations of these. Most of the experiments were carried out in the laboratory using 1‐year‐old seedlings or 1‐month‐old sprouts. Height and diameter growth, biomass production, morphology and ethylene evolution were studied for 1–2 months. Material for comparison was obtained from young sprouts on the stumps of fully developed trees growing under natural conditions and natural seedlings of a comparable age. Exposure of seedlings and coppiced plants to stress factors usually altered growth and increased ethylene evolution. Cutting of the stem and thinning had similar effects in that they stimulated transient ethylene evolution by both roots and stems. Bending retarded the height growth to some extent, while increasing the ethylene content especially on the lower side of the horizontal stem. Flooding arrested height growth and increased ethylene evolution in the roots. The two species reacted basically in similar ways, but B. pendula produced more ethylene. though at a slower rate, and also showed a more pronounced retardation of growth. The sprouts had a higher ethylene content than the seedlings and also increased their ethylene evolution more rapidly. The changes in ethylene evolution and growth are discussed from the point of view of the resilience of the plants and as an indication of stress factors in experimental plots.     

Leaves from grafted Meliaceae species affect survival and performance of <Emphasis Type="Italic">Hypsipyla grandella</Emphasis> (Zeller) (Lepidoptera: Pyralidae) larvae

In order to determine their effects on growth and mortality of instar II Hypsipyla grandella (Zeller), larvae were fed with leaf disks taken from shoots of susceptible species (Cedrela odorata L. and Swietenia macrophylla King) scions grafted onto resistant ones (Khaya senegalensis Desr. A. Juss and Toona ciliata M. Roem.), from their reciprocal grafts, and from both intact and autografted plants. In addition, crude leaf extracts from the susceptible and resistant plants, as well as from C. odorata grafted onto T. ciliata plants, were tested on C. odorata leaf disks. Mortality was evaluated 2, 10, and 25 days after starting the bioassay. Leaf area consumed and weight gain per larva were assessed 2 days after starting bioassay. Time to reach pupation, pupal weight and length 1 day after pupation, and time to adult stage and appearance of wings were determined at the end of the bioassay. Plant species significantly affected mortality (P ≤ 0.04) throughout the test. Eighty to 100% of larvae fed leaf disks from intact T. ciliata and its autograft, or C. odorata onto T. ciliata and its reciprocal graft died in the first 2 days of evaluation. All other factors measured, except pupal weight and length, were also affected (P ≤ 0.01) by the leaf disks. Intact resistant plants and reciprocal grafted plants reduced leaf consumption and caused larval weight loss. Larvae fed on K. senegalensis grafted onto S. macrophylla extended by 8 days the time to pupa and to adult stages and induced abnormal wing formation compared to larvae fed intact leaves of C. odorata. Crude extracts from resistant plants equally affected larval survival and performance compared to crude extract from C. odorata grafted onto T. ciliata plants, and these extracts were more detrimental to larvae than those from susceptible species. This study demonstrated that grafting borer-susceptible species on resistant rootstocks can affect the survival and performance of instar II H. grandella larvae.     

Responses of sugar maple and hemlock seedlings to elevated carbon dioxide under altered above- and belowground nitrogen sources

Various human-induced changes to the atmosphere have caused carbon dioxide (CO?), nitrogen dioxide (NO?) and nitrate deposition (NO??) to increase in many regions of the world. The goal of this study was to examine the simultaneous influence of these three factors on tree seedlings. We used open-top chambers to fumigate sugar maple (Acer saccharum) and eastern hemlock (Tsuga canadensis) with ambient or elevated CO? and NO? (elevated concentrations were 760 ppm and 40 ppb, respectively). In addition, we applied an artificial wet deposition of 30 kg ha?1 year?1 NO?? to half of the open-top chambers. After two growing seasons, hemlocks showed a stimulation of growth under elevated CO?, but the addition of elevated NO? or NO?? eliminated this effect. In contrast, sugar maple seedlings showed no growth enhancement under elevated CO? alone and decreased growth in the presence of NO? or NO??, and the combined treatments of elevated CO? with increased NO? or NO?? were similar to control plants. Elevated CO? induced changes in the leaf characteristics of both species, including decreased specific leaf area, decreased %N and increased C:N. The effects of elevated CO?, NO? and NO?? on growth were not additive and treatments that singly had no effect often modified the effects of other treatments. The growth of both maple and hemlock seedlings under the full combination of treatments (CO??+?NO??+?NO??) was similar to that of seedlings grown under control conditions, suggesting that models predicting increased seedling growth under future atmospheric conditions may be overestimating the growth and carbon storage potential of young trees.     

Effects of stem bark and leaf extracts of Bridelia ferruginea on rat bladder smooth muscle

The effects of ethanol extracts of Bridelia ferruginea leaves and stem bark on purinergic neurotransmission in the rat bladder were investigated. The stem bark extract potentiated the contraction of the bladder evoked by exogenous adenosine 5-triphosphate (ATP) but depressed KCl-induced contractions in a dose-related pattern; these two opposite actions might account for the lack of effect on field stimulation of the bladder purinergic nerves. The leaf extract depressed purinergic nerve-mediated contraction of the rat bladder in a dose-related fashion. This action could be attributed to blockade of purinergic neurotransmission since the leaf extract did not affect KCl-induced contractions.     

Effects of elevated CO(2) on chloroplast components, gas exchange and growth of oak and cherry

Specific chloroplast proteins, gas exchange and dry matter production in oak (Quercus robur L.) seedlings and clonal cherry (Prunus avium L. x pseudocerasus Lind.) plants were measured during 19 months of growth in climate-controlled greenhouses at ambient (350 vpm) or elevated (700 vpm) CO(2). In both species, the elevated CO(2) treatment increased the PPFD saturated-rate of photosynthesis and dry matter production. After two months at elevated CO(2), Prunus plants showed significant increases in leaf (55%) and stem (61%) dry mass but not in root dry mass. However, this initial stimulation was not sustained: treatment differences in net assimilation rate (A) and plant dry mass were less after 10 months of growth than after 2 months of growth, suggesting acclimation of A to elevated CO(2) in Prunus. In contrast, after 10 months of growth at elevated CO(2), leaf dry mass of Quercus increased (130%) along with shoot (356%) and root (219%) dry mass, and A was also twice that of plants grown and measured at ambient CO(2). The amounts of Rubisco and the thylakoid-bound protein cytochrome f were higher in Quercus plants grown for 19 months in elevated CO(2) than in control plants, whereas in Prunus there was less Rubisco in plants grown for 19 months in elevated CO(2) than in control plants. Exposure to elevated CO(2) for 10 months resulted in increased mean leaf area in both species and increased abaxial stomatal density in Quercus. There was no change in leaf epidermal cell size in either species in response to the elevated CO(2) treatment. The lack of acclimation of photosynthesis in oak grown at elevated CO(2) is discussed in relation to the production and allocation of dry matter. We propose that differences in carbohydrate utilization underlie the differing long-term CO(2) responses of the two species.     

南方红豆杉叶浸提液对小麦种子萌发与幼苗生长的影响

采用滤纸培养皿法研究了南方红豆杉叶浸提液对小麦种子萌发与幼苗生长的影响。研究结果发现,红豆杉叶浸提液对小麦种子的萌发和幼苗生长存在不同程度的抑制作用。低浓度的南方红豆杉浸提液促进小麦种子的萌发,高浓度浸提液抑制小麦种子的萌发。当浸提液浓度为6g/mL时,小麦种子的萌发率达到最高。南方红豆杉叶浸提液对小麦幼苗根系生长产生显著抑制作用。随着浸提液浓度的升高,小麦须根的长度逐渐下降,但不定根的数目逐渐增加。当浸提液的浓度为10g/mL时,根长抑制率达到42.48%。方差分析结果表明,南方红豆杉叶浸提液对小麦的根生长产生显著的抑制作用。对小麦幼苗的茎生长、根的生物量和茎的生物量均表现为低浓度的浸提液促进茎的生长和生物量的增加,高浓度浸提液则抑制茎的生长和生物量增加。研究结果表明,南方红豆杉浸提液表现低浓度时促进种子的萌发、须根数目的增加、茎的生物及生物量的增加,高浓度时则产生一定的抑制作用,而对根长的生长表现为显著的抑制作用。     

Arbuscular mycorrhizal induced changes to plant growth and root system morphology in Prunus cerasifera

We compared root system morphogenesis of micropropogated transplants of Prunus cerasifera L. inoculated with either of the arbuscular mycorrhizal (AM) fungi Glomus mosseae or Glomus intraradices or with the ericoid mycorrhizal species Hymenoscyphus ericae. All plants were grown in sand culture, irrigated with a nutrient solution that included a soluble source of phosphorus, for 75 days after transplanting. Arbuscular mycorrhizal colonization increased both the survival and growth (by over 100%) of transplants compared with either uninoculated controls or transplants inoculated with H. ericae. Arbuscular mycorrhizal colonization increased root, stem and leaf weights, leaf area, root length and specific leaf area, and it decreased root length/leaf area ratio, root/shoot weight ratio and specific root length. Both uptake of phosphorus and its concentration in leaves were increased by AM infection, although the time course of the relationships between intensity of AM infection and P nutrition were complex and suggested a role for factors other than nutrition. The time course for the development of infection varied. It was most rapid with G. mosseae, but it was ultimately higher with G. intraradices. None of the treatments significantly affected the lengths of adventitious roots or the first-, second- or third-order laterals that developed from them. Arbuscular mycorrhizal colonization increased the intensity of branching in all root orders with the effect being most obvious on first-order lateral roots where the number of branches increased from under 100 to over 300 brances m(-1). As a result, although first-order laterals made up 55% of the root systems of control plants, the comparable value was 36% in AM-infected plants. In contrast, second-order laterals represented 25% of control root systems, but 50% of AM-colonized root systems. Glomus intraradices but not G. mosseae increased root diameter. Anatomical studies revealed no changes in the overall form of the root tip, although there were changes in the diameter of the root cap, cell numbers and cell size. Hymenoscyphus ericae increased the duration of the metaphase index. Both AM fungal treatments increased the concentrations of soluble proteins in root extracts and modified the protein profiles by the elimination and addition of protein bands detected by PAGE analysis. We conclude that AM fungal inoculation influenced processes in the root system at different levels, but not all effects were due to improved P nutrition or increased physiological age.