Phenolic compounds accumulation in <Emphasis Type="Italic">Hypericum ternum</Emphasis> propagated in vitro and during plant development acclimatization

The phenolic compound content of Hypericum ternum was investigated after micropropagation establishment and during acclimatization over the phenological development of the plant. Plantlets cultured in vitro on full Murashige and Skoog medium without growth regulators displayed higher phenolic compound yields, were acclimatized, and field grown. Production of total phenolic compounds as well as hyperoside, chlorogenic acid, quercitrin, guaijaverin, isoquercitrin, and uliginosin B were quantified at vegetative, flowering and fructification stages, and different plant organs (roots, stems, leaves and reproductive parts) showing that reproductive parts at flowering stage and the leaves at fructification stage were the main repository site of secondary metabolites, except for uliginosin B. The stems were the least accumulative organ, while the roots accumulated only hyperoside and uliginosin B. Moreover, the accumulation of most of the flavonoids and uliginosin B in acclimatized plants surpassed the levels found in the wild plant, warranting further research with the species.     

The spatial pattern of leaf phenology and its response to climate change in China

Leaf phenology has been shown to be one of the most important indicators of the effects of climate change on biological systems. Few such studies have, however, been published detailing the relationship between phenology and climate change in Asian contexts. With the aim of quantifying species’ phenological responsiveness to temperature and deepening understandings of spatial patterns of phenological and climate change in China, this study analyzes the first leaf date (FLD) and the leaf coloring date (LCD) from datasets of four woody plant species, Robinia pseudoacacia, Ulmus pumila, Salix babylonica, and Melia azedarach, collected from 1963 to 2009 at 47 Chinese Phenological Observation Network (CPON) stations spread across China (from 21° to 50° N). The results of this study show that changes in temperatures in the range of 39–43 days preceding the date of FLD of these plants affected annual variations in FLD, while annual variations in temperature in the range of 71–85 days preceding LCD of these plants affected the date of LCD. Average temperature sensitivity of FLD and LCD for these plants was ?3.93 to 3.30 days °C^?1 and 2.11 to 4.43 days °C^?1, respectively. Temperature sensitivity of FLD was found to be stronger at lower latitudes or altitude as well as in more continental climates, while the response of LCD showed no consistent pattern. Within the context of significant warming across China during the study period, FLD was found to have advanced by 5.44 days from 1960 to 2009; over the same period, LCD was found to have been delayed by 4.56 days. These findings indicate that the length of the growing season of the four plant species studied was extended by a total of 10.00 days from 1960 to 2009. They also indicate that phenological response to climate is highly heterogeneous spatially.     

Damage by <Emphasis Type="Italic">Tibraca limbativentris</Emphasis> Stål (Pentatomidae) to Upland Rice Cultivated in Amazon Rainforest Region (Brazil) at Different Growth Stages

In this study, we evaluated the damage caused in the field by Tibraca limbativentris Stål adults at different levels of infestation (0, 1, 2, and 4 stink bugs) per three rice plants during three growth stages (V8, V13, and R4 stages) of upland rice cultivated in southwestern of Pará State, Amazon Rainforest region, Brazil. Heading time (panicle exertion) was affected by T. limbativentris infestations mainly in the vegetative stage and the whiteheads percentage in treatments ranged from 18.2 to 38%. The dead hearts percentages varied between 0 and 21.5%, and the mean number of primary branches (ramifications) ranged from 5.9?±?0.4 to 12.3?±?0.2. The number of empty spikelets was only affected in infestations with four insects/three plants, while the quantity of filled grains per panicle was affected only when infestations occurred during the vegetative stage. The total number of spikelets (filled?+?empty) per panicle decreased significantly in all phenological stages, and the percentage of damage ranged from 17 to 44% among treatments. Based on the proportion of damage observed, we suggest doubling the number of insects presently used as action threshold to 2 and 4 stink bugs per 15 stalks sampled for the vegetative stage, and of 1 or 2 stink bugs per 15 stalks sampled at the beginning of reproductive stage (R3/R4). Also, the field should be monitored during the entire vegetative stage, since most damage was observed in this phenological stage.     

Metabolic Alterations in Two Cirsium Species Identified at Distinct Phenological Stages using UPLC‐QTOF/MS

Introduction

Cirsium chanroenicum and C. setidens are commonly used both in traditional folk medicine and as a food source. The quality of different species of Cirsium at different harvest times is a function of their metabolite composition, which is determined by the phenological stage.

Objective

We sought to determine the differences in the metabolite composition of two species of Cirsium during different phenological stages using ultra‐performance liquid chromatography (UPLC) quadrupole time‐of‐flight (QTOF) mass spectrometry (MS).

Methodology

Cirsium chanroenicum and C. setidens plants were collected at the floral budding and full flowering stages. Metabolic profiles of Cirsium extracts were determined using UPLC‐QTOF/MS to characterise the differences between phenological stages, and the major metabolites were quantified using UPLC‐QTOF/MS‐multiple reaction monitoring (MRM).

Results

At the full flowering stage, the levels of phenolic acids as well as components of the phenylpropanoid pathway were increased. Flavonoids predominated at the full flowering stage in both species. The levels of coumaric acid, kaempferol, and pectolinarigenin differed between the two species of Cirsium. Overall, these results suggest that components of the phenylpropanoid metabolic pathway are upregulated in the full flowering stage in Cirsium, although we did observe some variation between the species.

Conclusion

These results will help elucidate the metabolic pathways related to the different phases of the vegetative cycle, and may help determine the optimal season for the harvest of Cirsium with the highest levels of bioactive compounds. Copyright © 2017 John Wiley & Sons, Ltd.     

Composition of volatile oils of Styrax (Styrax officinalis L.) leaves at different phenological stages

The volatile oil composition of Styrax officinalis L. leaves was investigated by GC and GC–MS during the phenological cycle of this species in a population growing wild in France. Whatever the analysed stage, yields were low 0.01–0.02%. (E)-2-hexenal, octanol and geraniol were the main constituents in all these volatile oils. The level of sesquiterpenoids was the highest at the vegetative stage. However, the chemical composition of leaf essential oil was relatively constant during the phenological stage of this tree and 58% of the identified compounds were common at all the analysed oils.     

Psychrophilic endophytic fungi with biological activity inhabit Cupressaceae plant family

Psychrophilic microorganisms are cold-adapted organisms that have an optimum growth temperature below 15 °C, and often below 5 °C. Endophytic microorganisms live inside healthy plants and biosynthesize an array of secondary metabolites which confer major ecological benefits to their host. We provide information, for the first time, on an endophytic association between bioactive psychrophilic fungi and trees in Cupressaceae plant family living in temperate to cold, semi-arid habitats. We have recovered psychrophilic endophytic fungi (PEF) from healthy foliar tissues of Cupressus arizonica, Cupressus sempervirens and Thuja orientalis (Cupressaceae, Coniferales). In total, 23 such fungi were found out of 110 endophytic fungal isolates. They were identified as ascomycetous fungi, more specifically Phoma herbarum, Phoma sp. and Dothideomycetes spp., all from Dothideomycetes. The optimal growth temperature for all these 23 fungal isolates was 4 °C, and the PEF isolates were able to biosynthesize secondary metabolite at this temperature. Extracted metabolites from PEF showed significant antiproliferative/cytotoxic, antifungal and antibacterial effects against phytopathogenic fungi and bacteria. Of special interest was their antibacterial activity against the ice-nucleation active bacterium Pseudomonas syringae. Accordingly, we suggest that evergreen Cupressaceae plants may benefit from their psychrophilic endophytic fungi during cold stress. Whether such endosymbionts confer any ecological and evolutionary benefits to their host plants remains to be investigated in vivo.     

Improving the use of aerobiological and phenoclimatological data to forecast the risk of late blight in a potato crop

The paper describes phenological and aerobiological monitoring conducted during 6 years on a potato crop. The progression of the phenological stages in relation with thermal time [growing degree-days (GDD) and physiological days (P-days)] was analyzed. The growing cycle for Kennebec variety required less than 120 days and mean values of 1700 GDD and 720 P-days to complete the phenological development. The presence of Phytophthora infestans in the environment was common in each crop cycle, but the maximum peaks of sporangia were detected a few days after the emergence of the plants. The growing cycles with lower maximum temperature had the highest presence of sporangia. Thus, significant negative correlations between mean temperature, maximum temperature, accumulated growing degree-days, accumulated physiological days and the concentration of P. infestans in the environment were found. Maximum temperature and accumulated growing degree-days of 5 previous days were the parameters that best estimated the P. Infestans concentration in the regression model applied, with an explained variance of the data of 33 and 34%.     

Influence of thermal requirement in the aerobiological and phenological behavior of two grapevine varieties

Knowledge about the timing of the aerobiological and phenological processes in plants with economic interest results of great agronomical importance, mainly for the establishment of the adaptive capacity of different varieties to various environmental conditions and the optimization of cultural practices. An agrometeorological study in two authorized varieties (Treixadura and Godello) of the Designation of Origin Ribeiro (North-western Spain) was conducted during the years 2008–2011. As consequence of the studied area bioclimatic conditions, the grapevine cycle exceed than 190 days and the varieties cultivated are considered as “late varieties.” The presence of Vitis pollen grains in the vineyard atmosphere was mainly registered during the stages 5 (inflorescence emerge) and 6 (flowering) which benefit a successful fertilization process. The variations in airborne pollen concentrations are related to temperature and humidity. Phenological models proposed in this paper offer a high accuracy as the standard deviation of error between estimated and observed values was low. The prediction variability ranges around 2 phenological scales in 2008 and 1 phenological scale in 2009, 2010 and 2011. These results indicate that the models developed to predict the phenology, in terms of degree days accumulated (GDD) using as a threshold temperature 10 °C, can be a useful tool to forecast the successive phenological events in the Designation of Origin Ribeiro area. During the stage 8 (ripening of berries), the GDD and the Brix Index were particularly correlated, so far as the differences between plants of the same variety are minimal.     

Growth kinetics and withanolide production in novel transformed roots of <Emphasis Type="Italic">Withania somnifera</Emphasis> and measurement of their antioxidant potential using chemiluminescence

Markedly increased withanolide content was found in transformed roots (TR) of Withania somnifera germplasm grown in low mineral minimal media and withanolides showed high antioxidant potential when analysed using acidic potassium permanganate chemiluminescence. Transformation frequency of explants infected with Agrobacterium rhizogenes strain A4 varied between the three germplasms tested with the highest observed as 75?±?0.9. Transformed root production was explant specific with leaves being the most productive among the different explants used. Withanolides, namely withaferin A, withanolide A, withanolide B and 12-deoxywithastramonolide were detected in TR cultures and differences in their content were found between germplasms. The highest concentrations of secondary metabolites were found in 4-week-old cultures and concentrations declined by the 8th and 12th week of culture. In 4-week-old cultures, the biomass of TR cultures was 4.5 fold higher than their respective non-transformed roots (NTR). Withaferin A was found in TR at levels that were 28–34 times higher than that found in NTR. A rapid method for the determination of the antioxidant potential of W. somnifera TR extracts was developed using post-column acidic potassium permanganate chemiluminescence (APPC) detection. The APPC chromatographic peaks for extract constituents showed strong alignment with those found for ultraviolet absorbance detection. The methods developed in this study for TR culture establishment and the use of a fast and sensitive way for the qualitative and quantitative determination of the antioxidant activity of their metabolites provides a new platform that will have use for similar studies in other species.     

Developmental Regulation of Low-temperature Tolerance in Winter Wheat

Vernalization and photoperiod genes have wide-ranging effectson the timing of gene expression in plants. The objectives ofthis study were to (1) determine if expression of low-temperature(LT) tolerance genes is developmentally regulated and (2) establishthe interrelationships among the developmental stages and LTtolerance gene expression. LT response curves were determinedfor three photoperiod-sensitive LT tolerant winter wheat (Triticumaestivum L. em Thell) genotypes acclimated at 4 °C under8 h short-day (SD) and 20 h long-day (LD) photoperiods from0 to 112 d. Also, three de-acclimation and re-acclimation cycleswere used that bridged the vegetative/reproductive transitionpoint for each LD and SD photoperiod treatment. A vernalizationperiod of 49 d at 4 °C was sufficient for all genotypesto reach vernalization saturation as measured by minimum finalleaf number (FLN) and confirmed by examination of shoot apicesdissected from crowns that had been de-acclimated at 20 °CLD. Before the vegetative/reproductive transition, both theLD- and SD-treated plants were able to re-acclimate to similarLT₅₀(temperature at which 50% of the plants are killed by LTstress) levels following de-acclimation at 20 °C. De-acclimationof LD plants after vernalization saturation resulted in rapidprogression to the reproductive phase and limited ability tore-acclimate. The comparative development of the SD (non-flowering-inductivephotoperiod) de-acclimated plants was greatly delayed relativeto LD plants, and this delay in development was reflected inthe ability of SD plants to re-acclimate to a lower temperature.These observations confirm the hypothesis that the point oftransition to the reproductive stage is pivotal in the expressionof LT tolerance genes, and the level and duration of LT acclimationare related to the stage of phenological development as regulatedby vernalization and photoperiod requirements. Copyright 2001Annals of Botany Company Triticum aestivum L., wheat, low-temperature tolerance, vernalization, photoperiod, phenological development     

Phenological development of natural populations of European field pansy (Viola arvensis) in a semi‐arid Mediterranean environment

The phenological stages, dry matter allocation and relative growth rate (RGR) of natural populations of European field pansy (Viola arvensis) were studied under field conditions for 3 years. The species exhibited an annual life cycle of 78–100 calendar days, depending on the growing season. The life cycle was initiated in early spring and ended by early summer, with an average accumulation of 1100 growing degree days. The initiation of emergence occurred in early spring at a range of maximum air temperatures between 15 and 20°C, coupled with a range of minimum air temperatures between 5 and 10°C. With increasing temperatures in mid‐spring, there was a rapid transition to the reproductive stage. Time to flowering was shortened considerably in the warm growing season. After flowering, the species appeared to allocate a relatively large proportion of biomass to the reproductive parts than dry matter allocation to the vegetative parts during juvenile stages. The maximum value of RGR was observed in the vegetative stage, whereas RGR values decreased sharply with plant age. Overall, the species showed a short life cycle and a reproductive strategy that directs resources to reproductive rather than vegetative output, thus increasing the likelihood of persistence in systematically disturbed environments that are characterized by limited growth time as a result of major events in the field, such as mechanical cultivation or herbicide application.     

Microbial secondary metabolites ameliorate growth, <Emphasis Type="Italic">in planta</Emphasis> contents and lignification in <Emphasis Type="Italic">Withania somnifera</Emphasis> (L.) Dunal

In the present investigation, metabolites of Streptomyces sp. MTN14 and Trichoderma harzianum ThU significantly enhanced biomass yield (3.58 and 3.48 fold respectively) in comparison to the control plants. The secondary metabolites treatments also showed significant augmentation (0.75–2.25 fold) in withanolide A, a plant secondary metabolite. Lignin deposition, total phenolic and flavonoid content in W. somnifera were maximally induced in treatment having T. harzianum metabolites. Also, Trichoderma and Streptomyces metabolites were found much better in invoking in planta contents and antioxidants compared with their live culture treatments. Therefore, identification of new molecular effectors from metabolites of efficient microbes may be used as biopesticide and biofertilizer for commercial production of W. somnifera globally.     

Warm temperature conditions restrict the sexual reproduction and vegetative growth of the spring ephemeral <Emphasis Type="Italic">Gagea lutea</Emphasis> (Liliaceae)

The responses of reproduction and growth to climate warming are important issues to predict the fate of plant populations at high latitudes. Spring ephemerals inhabiting cool-temperate forests grow better under cool conditions, but how reproductive performance is influenced by warm weather is unclear. The phenological and physiological responses of reproduction and vegetative growth to warm temperature and light conditions were evaluated in the spring ephemeral Gagea lutea. Leaf and bract physiological activities, bulb growth, and seed production were compared among reproductive plants grown in forest, open, and greenhouse (GH; warming manipulation in the open site) plots. In vitro pollen germination ability was tested under various temperatures. In the GH, leaf and bract photosynthetic activities decreased rapidly at the fruiting stage, but dark respiration rates remained high, resulting in higher carbon exhaust in warm conditions. Both leaf and bract sizes and their longevities were reduced in the GH. Annual bulb growth was largest in the forest plot and smallest in the GH plot. Pollen germination was strongly inhibited at high temperature (30 °C). Fruit and seed productions were decreased only in the GH plot. Both vegetative and reproductive activities were negatively affected by warm temperature, resulting in less vegetative growth and lower seed-set, whereas an understory habitat was beneficial for vegetative growth and showed similar seed production to an open habitat over the experimental period. Decreasing population dynamics of spring ephemerals was predicted in response to future warming climate not only by growth inhibition but also by restriction of seed production.     

Effects of salt marsh invasion by Spartina alterniflora on sulfate-reducing bacteria in the Yangtze River estuary,China

To investigate how plant invasion affects sulfate-reducing bacteria (SRB) responsible for sulfate reduction, we conducted a comparative study of diversity and composition of SRB in rhizosphere soils of invasive exotic species (Spartina alterniflora) and two native species (Phragmites australis and Scirpus mariqueter) on Jiuduansha Island located in the Yangtze River estuary, China. Throughout the growing season, profiles of DGGE fingerprints of SRB had distinct variations in relation to phenological stages of these three plant species. The higher richness and abundance of SRB in the rhizospheres of native plants mainly occurred when the plants were in vegetative growth and reproductive stages. However, the higher richness and abundance of SRB also occurred in the late growing season (senescent stage) of S. alterniflora rhizosphere, during which Desulfobulbus, Desulfuromonas, Desulfovibrio, and Firmicutes were dominant. Our results adding to our previous studies suggested that abundant SRB in late stage might have close relationships with decomposition of soil organic matters produced by S. alterniflora.     

Metabolic signatures altered by in vitro temperature stress in <Emphasis Type="Italic">Ajuga bracteosa</Emphasis> Wall. ex. Benth.

To elucidate how biosynthesis of plant metabolites is affected by temperature, metabolite profiles from in vitro regenerated plants raised under different temperature regimes of 10, 15 °C, 20 °C, 25 °C and 30 °C were obtained using electrospray ionization mass spectrometry (ESI-MS), and principal component analysis (PCA) was carried out to identify key metabolites. Several bin masses were detected by PCA loading scatter plots which separated the samples. In-house bin program selectively manifested the putative known metabolites depending on % total ions count and intensity of selected bins in the plant samples. Total phenolic and flavonoid content were harvested to highest levels (12.9 mg GAE/g DW and 9.3 mg QE/g DW), respectively, at 15 °C. Besides, pinoresinol (lignan), some of the vital amino acids such as serine, methionine, histidine and glutamine were found to be at higher amount in plants raised at 15 °C. Significant phenylpropanoids like cinnamic acid, caffeic acid and quercitol were detected at a higher concentration in plants raised at 15 °C as compared to other treatments. However, phosphoenolpyruvate, and oxalosuccinate (intermediates of the pentose phosphate pathway) were accumulated the most in plants raised at 30 °C and they were detected with lowest values at 10 °C. Glucose and deoxy-xylose 5 phosphate (intermediates of TCA cycle) were found in higher amounts at temperature treatments of 15 and 25 °C, respectively. We conclude that a low-temperature treatment (15 °C) results in a stress-induced accumulation of a variety of pharmacologically important secondary metabolites.     

Developmental regulation of metabolites and low temperature tolerance in lines of crosses between spring and winter wheat

Low temperature (LT) tolerance in cereals needs developmental regulation of metabolites, a process which is associated with vernalization requirement. This study was initiated to investigate the relationships among stage of phenological development, final leaf number (FLN), the activities superoxide dismutase, catalase, guaiacol peroxidase, ascorbate peroxidase and polyphenol oxidase, the contents of proline, photosynthetic pigments, and hydrogen peroxide (H₂O₂) during vernalization and LT acclimation in spring and winter wheat. Six genotypes with different vernalization requirements were grown under greenhouse and field conditions. The spring-habit parent, “Pishtaz” and line 4021, rapidly entered the reproductive phase and had a limited capacities to LT acclimate. They also had the lowest antioxidative activities and accumulation of proline among genotypes. Lines 4002 and 4014, with a short vernalization requirement and higher FLN, remained in the early stages of phenological development longer and developed a higher level of LT tolerance and metabolites compared to spring habit genotypes. In contrast, the winter habit “Norstar” and line 4023 spent a longer time in the vegetative stage and accumulated higher levels of metabolites. Maximum LT tolerance and metabolite accumulations occurred near the vegetative/reproductive transition in all genotypes. The longer periods of vernalization and increased FLN that happened along with increased defense mechanisms and decreased damage indices (H₂O₂ content and LT₅₀) ensured LT tolerance in wheat. These results demonstrate that both genetic and environmental factors via developmental regulation of metabolites play important roles in creating LT tolerance in long mild winters of Iran. Significant correlations coefficients for many of the metabolites considered in this study and Lethal temperature 50 (LT₅₀) also suggest that they could be useful as indirect measures of plant LT tolerance potential in wheat breeding programs.     

Arbuscular mycorrhizal fungal diversity in high-altitude hypersaline Andean wetlands studied by 454-sequencing and morphological approaches

Withania somnifera, also known as Indian ginseng is known to contain valuable bioactive compounds, called withanolides that structurally resemble ginsenosides of Panax ginseng. These compounds provide the basis of pharmacological relevance in traditional systems of medicine. In the present study, 150 hairy root lines of W. somnifera were induced of which nine fast growing lines were analysed for their growth and withanolide content. Hairy root line W9 was selected due to its high specific growth rate (0.196 ± 0.005 d^?1) and high withanolide content. The response to different concentrations of elicitors (methyl jasmonate and P. indica cell homogenate) and various exposure durations was assessed in the W9 hairy root line. The withanolide content as well as the pattern of gene expression from MVA, MEP and sterol pathway, was evaluated using qPCR. Though gene expression and withanolide content were found to be elevated in almost all MeJ and CHP treatments, the exposure of hairy roots to 15 μM MeJ for 4 h gave the maximum withanolide yield. The results suggest that the elicitation potential of methyl jasmonate was higher than that of P. indica cell homogenate for increasing withanolide levels in hairy roots of W. somnifera.     

Suppression of the source activity affects carbon distribution and frost hardiness of vegetating winter wheat plants

Effect of suppression of the source activity on some physiological characteristics of winter wheat (Triticum aestivum L., cv. Mironovskaya 808) was studied on plants grown in water culture. The plants were examined at the mixotrophic stage of growth period, during their transition from vegetative state to relative dormancy in autumn. The average temperature over 10 days of the experiment was 6°C at 9-h photoperiod and illuminance of 8–20 klx. The source strength was suppressed successively with a series of treatments: intact control plants (V1); plants with the seed endosperm removed (V2); plants with photosynthesis inhibited (V3); plants with the seed endosperm removed and photosynthesis inhibited (V4); plants with the seed endosperm removed, photosynthesis inhibited, and the root nutrient medium replaced with distilled water (V5). On the 6th–10th day of the experiment, the relative growth rate (RGR) was determined from dry weight increments. At the same time, the distribution of biomass among organs, the CO₂ exchange rates (photosynthesis and dark respiration), the content and proportions of sugars (sucrose, glucose, and fructose), the total content of phenolic compounds and flavonoids, the index of membrane stability (IMS) in leaves, and frost hardiness of plants were measured. Frost hardiness of vegetating plants was shown to be inversely related to RGR (R = ?0.906), dark respiration rate (R = ?0.789), the percentage of sucrose in total sugar content (R = ?0.737), leaf IMS (R = ?0.390), and the rate of apparent photosynthesis (R = ?0.288); it was directly proportional to the content of flavonoids (R =0.973), total phenols (R = 0.743), and sugars (R = 0.385). The role of modified source-sink relations in frost hardiness of vegetating plants at the stage of their transition to cold hardening is discussed. The differences between plants undergoing this transition and cold-hardened plants are considered, as well as the importance of phenolic compounds for the development of frost hardiness.     

Pseudohypericin and hyperforin in Hypericum perforatum from Northern Turkey: variation among populations, plant parts and phenological stages

Hypericum perforatum is a perennial medicinal plant known as "St. John's wort" in Western Europe and has been used in the treatment of several diseases for centuries. In the present study, morphologic, phenologic and population variability in pseudohypericin and hyperforin concentrations among H. perforatum populations from Northern Turkey was investigated for the first time. The aerial parts of H. perforatum plants representing a total of 30 individuals were collected at full flowering from 10 sites of Northern Turkey to search the regional variation in the secondary metabolits concentrations. For morphologic and phenologic sampling, plants from one site were gathered in five phenological stages vegetative,floral budding, full flowering, fresh fruiting and mature fruiting. The plant materials were air-dried at room temperature and subsequently assayed for chemical concentrations by high performance liquid chromatography. Secondary metabolite concentrations ranged from traces to 2.94mg/g dry weight (DW) for pseudohypedcin and traces -6.29mg/g DW for hyperforin. The differences in the secondary metabolite concentrations among populations of H. perforatum were found to be significant. The populations varied greatly in hyperforin concentrations, whereas they produced a similar amount of pseudohypericin. Concentrations of both secondary metabolites in all tissues increased with advancing of plant development and higher accumulation levels were reached at flowering. Among different tissues, full opened flowers were found to be superior to stems, leaves and the other reproductive parts with regard to pseudohypericin and hyperforin accumulations. The present findings might be useful to optimize the processing methodology of wild-harvested plant material and obtain Increased concentrations of these secondary metabolites.