Variation of Bioactive Compounds in Hypericum perforatum Growing in Turkey During Its Phenological Cycle

The present study was conducted to determine phenologic and morphogeneUc variation of hyperlcln, chlorogenlc acid and flavonoids, as rutin, hyperoside, apigenin-7-O-glucoside, quercitrin, quercetin content of Hypericum perforatum L. growing in Turkey. Wild growing plants were harvested at vegetative, floral budding, full flowering, fresh frulUng and mature fruiting stages and dissected into stem, leaf and reproductive tissues and assayed for bioacUve compounds by the High performance liquid chromatography （HPLC） method. Hypericin concentration ranged between 0 and 2.73 mg/g DW, chlorogenic acid 0.00-3.64 mg/g DW, rutin 0.00-3.36 mg/g DW, hyperoside 0.04- 22.42 mg/g DW, quercitrin 0.03-3.46 mg/g DW and quercetin 0.04-1.02 mg/g DW depending on ontogenetic and morphogenetic sampling. Leaves were found to be superior to stems and reproductive parts with regard to phenolic accumulation for all compounds tested while flowers accumulated the highest levels of hypericln. Quercltrln, quercetln and hypericin content in all tissues increased with advancing of developmental stages and reached their highest level during flower ontogenesis. Similarly, chlorogenic acid, hyperoside and apigenin-7-O-glucoside content in different plant parts increased during plant development, however, the highest level was observed at different stages of plant phenology for each tissue. Chlorogenic acid was not detected in stems, leaves and reproductive parts in several stages of plant phenology and its variation during plant growth showed inconsistent manner. In contrast to the other compounds examined, rutin content of stems and leaves decreased with advanc- ing of plant development and the highest level for both tissues was observed at the vegetative stage. However, content of the same compound in reproductive parts was the highest at mature fruiting. The present findings might be useful to obtain increased concentration of these natural compounds.     

Variation of bioactive secondary metabolites in <Emphasis Type="Italic">Hypericum origanifolium</Emphasis> during its phenological cycle

The genus Hypericum has received considerable interest from scientists, as it contains the variety of structurally diverse natural products which possess a wide array of biological properties. The present study was conducted to determine ontogenetic and morphogenetic variation of hypericin, chlorogenic acid and flavonoids, as rutin, hyperoside, apigenin-7-O-glucoside, quercitrin and quercetin content in Hypericum origanifolium growing in Turkey. Wild growing plants were harvested at vegetative, floral budding, full flowering, fresh fruiting and mature fruiting stages and dissected into stem, leaf and reproductive tissues and assayed for bioactive compounds by HPLC method. Hypericin, quercetin and quercitrin content in whole plant increased during course of ontogenesis and the highest level was reached in blooming stage. On the contrary, hyperoside content of whole plant decreased linearly with advancing of development stages and the highest level was observed at vegetative stage. Plants produced similar amount of chlorogenic acid at all stages of plant phenology except for mature fruiting at which the amount of this compound was decreased sharply. Among different tissues, reproductive parts accumulated the highest level of hypericin, quercetin and quercitrin, however, leaves produced substantially higher amount of chlorogenic acid and hyperoside. Rutin and apigenin-7-O-glucoside were detectable in all tissues only during fruit maturation. The presence and variation of these bioactive substances in H. origanifolium were reported for the first time.     

Coal-smoke pollution modifies physio-chemical characteristics of tissues during the ontogeny of <Emphasis Type="Italic">Peristrophe bicalyculata</Emphasis>

Coal-smoke emissions of a thermal power plant affected the physio-chemical status of Peristrophe bicalyculata (Reth) Nees, as observed at the pre-flowering, flowering and post-flowering stages of plant growth. The nitrate level was raised while nitrate reductase activity, and the soluble protein content of leaf declined heavily at the polluted site during different stages of plant growth, compared to the control. The rate of photosynthesis also decreased under the pollution stress. Sugar level in root, stem and leaves increased with growing age of the plant but was always lower at the polluted site than at the reference site. In roots, the difference was marginal till flowering stage and quite conspicuous afterwards; stems showed a reverse pattern of variation. Sulphur content was higher at the polluted site in all the organs and at each stage of the plant life. The Zn and Fe concentrations were reduced in all plant parts under the pollution stress. Copper content in root was consistently low at the polluted site. In the stem and leaves, however, it was almost equal on both the sites at the pre-flowering stage but showed a wide difference during the later part of plant ontogeny.     

Resource allocation in different parts of juvenile mountain birch plants: effect of nitrogen supply on seedling phenolics and growth

The composition and concentrations of phenolic compounds were studied in the first true leaves, cotyledons, stems and roots of 2.5-week-old seedlings of mountain birch ( Betula pubescens ssp. czerepanovii ). The differences in secondary compounds among these plant parts were both qualitative and quantitative. In all parts, condensed tannins accounted for more than 50% of the phenolics. In the first true leaves and cotyledons, chlorogenic acid was the most abundant of the HPLC phenolics. The main components in stems were (+)-catechins and rhododendrins whereas in roots, the main components were ellagitannins. The seedlings were grown at three levels of nitrogen supply (very low-N, low-N, moderate-N), and the effect of nitrogen on concentrations of phenolic compounds was studied in all plant parts. The dry weight of all plant parts, except the roots, increased with increased nitrogen. In all parts, the concentration of condensed tannins was higher at lower levels of nitrogen than at moderate-N. The concentrations of total HPLC phenolics and also those of the compound groups of HPLC phenolics were, however, affected only in the first true leaves and roots. The concentrations in the first true leaves were generally higher in seedlings grown at very low-N and low-N than in seedlings grown at moderate-N. The concentrations in roots were highest at low-N. Not all compounds responded to nitrogen supply in the same manner. The changes in concentrations cannot be exclusively interpreted as changes in the accumulation of phenolic compounds, due to dilution caused by the increase in biomass in better nitrogen availability. There were differences in carbon allocation between condensed tannins and HPLC phenolics in seedlings grown at different nitrogen levels.     

Benzopyrans in <Emphasis Type="Italic">Hypericum polyanthemum</Emphasis> Klotzsch ex Reichardt cultured in vitro

Hypericum polyanthemum Klotzsch ex Reichardt, an endemic species of Southern Brazil, was micropropagated on MS medium supplemented with 1.78 μM BAP. Shoot proliferation and rooting was achieved on hormone-free medium and plantlets survived acclimatization. The bioactive compounds: 6-isobutyryl-5,7-dimethoxy-2,2-dimethyl-benzopyran (HP1), 7-hydroxy-6-isobutyryl-5-methoxy-2,2-dimethyl-benzopyran (HP2) and 5-hydroxy-6-isobutyryl-7-methoxy-2,2-dimethyl-benzopyran (HP3) were quantified in the leaves, stems and roots of propagated and acclimatized plantlets and compared with the field-grown plants. The HPLC analysis revealed that the three benzopyrans are accumulated in the aerial parts and the concentration varied with the age of the plant whereas the roots were capable of accumulating only HP3. Greatest yield of HP1 (7.12 mg/g DW) was quantified in the leaves of the acclimatized plantlets, whereas the flowers of the plants from natural habitat displayed higher amounts of HP2 (11.04 mg/g DW) and HP3 (13.99 mg/g DW).     

Photosynthesis,dry matter accumulation and distribution in the wild sunflower Helianthus petiolaris and the cultivated sunflower Helianthus annuus as influenced by water deficits

Summary This study reports on the effects of water deficits on photosynthesis, plant growth and carbon allocation in the wild sunflower Helianthus petiolaris and in the cultivated sunflower Helianthus annuus grown under controlled conditions in the glasshouse. Water deficits reduced the rate of net photosynthesis and the dry weight of leaves, stems, roots and reproductive parts in both species. The root-to-shoot ratio of about 0.05 in H. petiolaris was lower than the root-to-shoot ratio of about 0.15 in H. annuus. Water stress did not affect the root-to-shoot ratio, but increased the percentage of roots at depth in H. annuus. The decrease in growth induced by water deficits was a consequence of a reduction in both leaf area production and net photosynthesis. Flowering occurred earlier in H. petiolaris than in H. annuus with a consequent earlier allocation of carbon to reproductive parts in the wild compared to the cultivated sunflower. The time to budding and flowering of either species was not altered by mild water stress, but was delayed by severe water deficits. During mild water stress carbon allocation to stems decreased, but that to reproductive parts increased. When plants were severely stressed and then rewatered the proportion af carbon allocated to leaves increased and the proportion allocated to stems decreased when compared to unstressed plants. The adaptative role of these features is discussed.     

Zinc and Copper Bioaccumulation in Brassica napus at Flowering and Maturation

The aim of this research was to investigate the capability of Brassica napus to bioaccumulate zinc and copper from artificially contaminated soil at the flowering and maturation phenological stage. The trial was set up in a greenhouse and the plants were cultivated in pots. The agricultural soil utilized was contaminated with zinc and copper sulfate (300 and 600 mg/kg, respectively). The soil and plant samples were simultaneously collected during the flowering (8 weeks after seeding) and maturation (harvest time, 13 weeks after seeding) stages, and the heavy metal concentrations were then analyzed. The production of vegetable biomass and the length of the roots were measured. The results showed that B. napus accumulated zinc and copper and translocated these elements in different ways in the harvestable parts of the plants. The zinc bioaccumulation was higher than that of copper. At flowering, zinc was mainly accumulated in the shoots (stems + leaves). Copper was particularly accumulated in the roots, during the entire life cycle. Taking into account the biomass production, the highest heavy metal removal by the shoots (above‐ground parts) occurred at the flowering stage for both zinc and copper. The high bioavailability of zinc and copper in the soil did not severely affect the root length and the biomass production.     

Composition of the essential oils in various organs at different developmental stages of Ammi visnaga (L.) Lam. from Tunisia

The composition of the essential oils isolated by hydrodistillation from various organs at different development stages of Ammi visnaga (L.) Lam. growing in Tunisia was determined by GC/MS analysis. In particular, the oil profiles of the leaves, stems, flower buds, roots, umbels, and fruits have been examined during the whole life cycle. The oil from the flowering aerial parts was characterized by a high content of isoamyl 2-methylbutanoate. After flowering and during desiccation and fructification, the umbels and fruits expressed a high content of linalool. The oils, extracted from the roots collected in the vegetatif, buds floral, and floral stages, were rich in monoterpene aldehydes, oxygenated monoterpenes, and monoterpene hydrocarbons. The highest level of non-terpene hydrocarbons was found at the flower-bud stage, represented by 61.3% of nonane. Among the monoterpenes, sabinene (12.5%) and β-pinene (8.5%) were identified in the flower buds.     

Influence of [S, S]-EDDS on phytoextraction of copper and zinc by Elsholtzia splendens from metal-contaminated soil

Two pot experiments were conducted to investigate the time course effects of the (S, S)-N, N'-ethylenediamine disuccinic acid (EDDS) addition to contaminated soil on the uptake of Cu and Zn by the Cu accumulator Elsholtzia splendens and on plant Cu and Zn concentrations at different growth stages. EDDS increased the amounts of Cu and Zn soluble in the soil, taken up by plants, concentrated in the xylem sap, and translocated from roots to stems and leaves. The increase in soil-soluble metals, especially Cu, resulted in a corresponding increase in metal concentrations in the xylem sap and leaves. The addition of EDDS to the soil increased plant Cu and Zn concentrations, especially in the leaves, and changed the proportions of Cu and Zn taken up by different plant parts. The proportions of Cu and Zn taken up by the roots were higher than by the leaves of control plants, but EDDS-treated plants showed the opposite trend. EDDS exerted greater effects at the end of the vegetative growth stage than at the start of the flowering or reproductive stages.     

Allometric Effects of <Emphasis Type="Italic">Agriophyllum squarrosum</Emphasis> in Response to Soil Nutrients,Water, and Population Density in the Horqin Sandy Land of China

We monitored the allometric effects for greenhouse-grown Agriophyllum squarrosum plants in response to variations in population density and the availability of soil nutrients and water. Biomass allocations were size-dependent. The plasticity of roots, stems, leaves, and reproductive effort was “true” in response to changes in nutrient content. At a low level of soil minerals, plants allocated more resources to the development of roots and reproductive organs than to leaves, but data for stem allocations were consistent for tradeoffs between the effects of nutrients and plant size. The plasticities of leaf allocation and reproductive effort were “true” whereas those of root and stem allocations were “apparent” in response to fluctuations in soil water, being a function of plant size. Decreasing soil water content was associated with higher leaf allocation and lower reproductive effort. Except for this “apparent” plasticity of leaf allocation, none was detected with population density on biomass allocation. Architectural traits were determinants of the latter. For roots, the determining trait was the ratio of plant height to total biomass; for stems and reproduction, plant height; and for leaves, the ratio of branch numbers to plant height.     

Influence of fungal elicitation with Nomuraea rileyi (Farlow) Samson in the metabolism of acclimatized plants of Hypericum polyanthemum Klotzsech ex Reichardt (Guttiferae)

Changes in biomass and bioactive metabolites after treatment with the fungus Nomuraea rileyi (Farlow) Samson added as dried culture (DC) or as dried autoclaved cell powder (DACP) for short and long periods of time have been investigated in Hypericum polyanthemum plants grown under controlled conditions and after 18 weeks of field acclimatization. Plants grown under controlled conditions and treated with DACP showed increased concentrations of the benzopyrans HP1 (6-isobutyryl-5,7-dimethoxy-2,2-dimethylbenzopyran), HP2 (7-hydroxy-6-isobutyryl-5-methoxy-2,2-dimethylbenzopyran), and HP3 (5-hydroxy-6-isobutyryl-7-methoxy-2,2-dimethylbenzopyran) while DC affected negatively or did not alter the synthesis of these compounds. Lower levels of the phloroglucinol derivative uliginosin B were detected in all treatments. Long time treatment with DACP of acclimatized plants triggered doubling of biomass as compared to control and chemical analyses demonstrated significant increase on total phenolic compounds in vegetative parts of plants. Moreover, the higher performance liquid chromatography analysis showed different pattern of metabolites accumulation, with higher yields of HP1, HP2, HP3 and uliginosin B in the reproductive parts of the plants treated with DACP during all experiments. The elevation of bioactive metabolites levels in response to the elicitor suggests that these compounds are inducible as part of the defense response of H. polyanthemum.     

Morphological and anatomical variations ofCajanus cajan (Linn.) huth raised in cadmium-rich soil

Different concentrations of cadmium in the growing media affected morphological parameters ofCajanus cajan. Over time, the amount of increase in shoot and root lengths, number of branches and leaves per plant, single and total leaf areas, and dry mass of leaves, was significantly lower for treated plants compared with controls. The root-shoot length ratio, which varied little over time, was relatively low for the treated plants. Although dry mass of both stems and roots increased, the rates were considerably low under Cd stress. The root-shoot dry mass ratio in the controls was highest during flowering and lowest in the post-flowering stage, but continually declined over time for the stressed plants. Compared with the controls, treated plants had fewer pods, with the number decreasing as the Cd concentration increased. Cd content was greater in roots than in stems or leaves, and leaves had greater amounts than did stems at higher doses. For all plants, the width and density of vessel elements and the length of fibers in the wood of stems and roots increased with plant age. However, the rate of increase was generally lower in the treated plants, the difference being more pronounced with higher doses of Cd. This indicated a reduced ascent of sap and, hence, less available water for tissues in treated plants.     

Reduction of ambient UV-B radiation does not affect growth but may change the flowering pattern of Rosmarinus officinalis L.

The effects of sub-ambient levels of UV-B radiation on the shrub Rosmarinus officinalis L. were investigated in a field filtration experiment in which the ambient UV-B was manipulated by a combination of UV-B transmitting and UV-B absorbing filters. As a result, the plants were receiving near-ambient or drastically reduced UV-B radiation doses. Drastic reduction of UV-B radiation had no effect on mean, total and maximum stem length, number of stems per plant, dry mass of leaves, stems and roots and leaf nitrogen and phenolic contents. However, flowering was more pronounced under reduced UV-B radiation during the winter period which coincides with ascending ambient UV-B radiation. In contrast, during autumn and early winter, a period which coincides with descending ambient UV-B radiation, flowering was unaffected by reduced UV-B radiation. We can conclude that natural UV-B radiation does not affect growth of Rosmarinus officinalis, but its reduction could influence the flowering pattern of the species.     

Effect of inoculation method and plant growth medium on endophytic colonization of sorghum by the entomopathogenic fungus <Emphasis Type="Italic">Beauveria bassiana</Emphasis>

A study was conducted to determine the effect of inoculation method and plant growth medium on colonization of sorghum by an endophytic Beauveria bassiana. Colonization of leaves, stems, and roots by B. bassiana was assessed 20-days after application of the fungus. Although B. bassiana established as an endophyte in sorghum leaves, stems, and roots regardless of inoculation method (leaf, seed, or soil inoculation), plant growth medium (sterile soil, non-sterile soil, or vermiculite) apparently influenced colonization rates. Seed inoculation with conidia caused no stem or leaf colonization by the fungus in non-sterile soil but did result in substantial endophytic colonization in vermiculite and sterile soil. Leaf inoculation did not result in root colonization, regardless of plant growth medium. Endophytic colonization was greater in leaves and stems than roots. Endophytic colonization by B. bassiana had no adverse effects on the growth of sorghum plants. Leaf inoculation with a conidial suspension proved to be the best method to introduce B. bassiana into sorghum leaves for plants growing in either sterile or non-sterile soil. Further research should focus on the virulence of endophytic B. bassiana against sorghum stem borers.     

Olfactory response of Trigonotylus caelestialium (Het.: Miridae) to rice plant and gramineous weeds

Abstract:  The olfactory response of Trigonotylus caelestialium , to rice, Oryza sativa L., and two species of gramineous weeds, Poa annua and Digitaria ciliaris , was investigated with an olfactometer to clarify the role of host-plant odours as olfactory cues in their host-finding behaviour. Third-instar nymphs and adult males were significantly attracted to whole plants (above ground parts) of P. annua . However, adult females were not significantly attracted to whole plants of P. annua . Attractancy of rice to T. caelestialium differed with the growth stage and part of the plant. Adult females were significantly attracted to stems and leaves in the panicle-formation stage, and panicles in the flowering stage. They were not attracted to stems and leaves in the fourth-leaf stage and flowering stage, or to panicles in the milk-and full-ripe stages. Although adult males were significantly attracted to stems and leaves in the panicle-formation stage, they were not attracted to the other rice structures tested. Both females and males were significantly attracted to stems and leaves, as well as panicles of D. ciliaris in the flowering stage. The findings suggest that T. caelestialium use host-plant volatiles as olfactory cues to find their host plants and their invasion to paddy fields were caused by olfactory responses to rice plant.     

Phenology of glucosinolate concentrations in roots,stems and leaves of Cardamine cordifolia

Total concentrations of isothiocyanate-yielding glucosinolates (IYG) were measured in roots, stems, basal leaves and cauline leaves of the herbaceous perennial Cardamine cordifolia (bittercress, Cruciferae), sampled at three sites in the Colorado Rockies during 1981. Significant variation in quantity was partitioned among plant parts, among sampling dates throughout the growing season, and among the three sites. Roots and basal leaves maintained high and similar concentrations of IYG through the season, while cauline leaves and stems showed seasonal declines, associated partly with flowering. Roots also consistently produced oxazolidinethione-yielding glucosinolates (hydroxylated analogues of IYG), whereas above-ground parts were variable for the presence of these compounds. Seasonal and plant-part variability in glucosinolate content and spatial patchiness of glucosinolate phenotypes contribute to the variation in herbivore occurrence and damage documented in previous studies of this native crucifer.     

基于广泛靶向代谢组学分析黑老虎不同部位成分差异

为提高黑老虎(Kadsura coccinea)资源的综合利用率，采用广泛靶向代谢组学技术鉴定并分析了根、茎、叶代谢组分差异及高度富集成分。结果表明，在根、茎和叶中分别鉴定出642、650和619个代谢物，以酚酸、脂质、类黄酮和有机酸为主；叶与根、茎与根的共有成分分别为566和650个，显著差异成分有442和393个，主要为酚酸、类黄酮和脂质，差异代谢物在苯丙烷生物合成、黄酮与黄酮醇生物合成通路中显著富集。代谢物总丰度和次生代谢物丰度均表现为叶>根>茎，叶中酚酸、类黄酮和脂质及茎中酚酸积累量显著高于根，而氨基酸及其衍生物、萜类、木脂素、香豆素、生物碱的丰度在根中显著上调。因此，黑老虎根、茎、叶有大量共有成分，叶和茎中酚酸、叶中类黄酮和脂质高度富集，含有新绿原酸、绿原酸、槲皮素等多个丰度较高且具有重要生物活性化合物，具有较高利用价值。     

Time course study on accumulation of cell wall-bound phenolics and activities of defense enzymes in tomato roots in relation to <Emphasis Type="Italic">Fusarium</Emphasis> wilt

Major cell wall-bound phenolic compounds were detected and identified in roots of tomato at different stages of growth. Alkaline hydrolysis of the cell wall material of the root tissues yielded ferulic acid as the major bulk of the phenolic compounds. Other phenolic compounds identified were 4-hydroxybenzoic acid, vanillic acid, 4-hydroxybenzaldehyde, vanillin and 4-coumaric acid. All the six phenolic acids were higher in very early stage of plant growth. Ferulic acid, 4-hydroxybenzoic acid and 4-coumaric acid exhibited a decreasing trend up to 60 days and then the content of these phenolic acids increased somewhat steadily towards the later stage of growth. Total phenolics, phenylalanine ammonia-lyase (PAL) activity and peroxidase (POD) activity were in tandem match with the occurrence pattern of the phenolic acids. Ferulic acid showed highest antifungal activity against tomato wilt pathogen Fusarium oxysporum f. sp. lycopersici. The results of this study may be interpreted to seek an explanation for high susceptibility of tomato plants at flowering stage to Fusarium wilt. It may also be concluded that greater amounts of ferulic acid in combination with other phenolics and higher level of PAL and POD activities after 60 days of growth may have a role in imparting resistance against Fusarium wilt at a late stage of plant growth.     

Initial burst of root development with decreasing respiratory carbon cost in Fagus crenata Blume seedlings

As terrestrial plants are rooted in one place, their metabolism must be acclimatized to continuously changing environmental conditions. This process is influenced by different metabolic traits of plant organs during ontogeny. However, direct measurement of organ-specific metabolic rates is particularly scarce, and little is known about their roles in whole-plant metabolism. In this study, we investigated size scaling of respiration rate, fresh mass and surface area of leaves, stems and roots in 65 seedlings of Fagus crenata Blume (2 weeks to 16 months old). With the increase in plant mass, the proportion of roots in whole plants increased from 20.8 to 87.3% in fresh mass and from 12.8 to 95.0% in surface area, whereas there was only a 15.6 to 60.2% increase in respiration rate. As a result, the fresh-mass-specific and surface-area-specific respiration rates in the roots decreased by 85% and 90%, respectively, and these decreases were significantly size dependent. However, such a size-dependent decrease was not observed for the surface-area-specific respiration rate in the leaves and stems. It is likely that this rapid root development is specific to the early growth stage after germination and would help plants acquire water and nutrients efficiently (i.e., at relatively low respiratory carbon costs). Overall, it is probable that the establishment of F. crenata forests and survival of F. crenata seedlings could be promoted by substantial root growth, with a reduction in respiratory carbon cost.