Effects of both substrate and nitrogen and phosphorus fertilizer on <Emphasis Type="Italic">Sphagnum palustre</Emphasis> growth in subtropical high-mountain regions and implications for peatland recovery

Human activities have recently caused severe destruction of Sphagnum wetlands in subtropical high-mountain regions, calling for urgent efforts to restore Sphagnum wetlands. Through a greenhouse experiment in western Hubei, China, we studied the effects of different substrate types (peat and mountain soil) and different levels of nitrogen (N) (0, 2, 4, 6, 10 g m^?2 year^?1) and phosphorus (P) (0, 0.2, 0.5, 1, 2 g m^?2 year^?1) on the growth of Sphagnum palustre, which was evaluated by four growth indicators: length growth, number of capitula, coverage change and biomass. We aimed to determine the optimal nutrient conditions for S. palustre growth, which would contribute to the rapid colonization and restoration of Sphagnum wetlands. The results showed that the different substrates significantly influenced S. palustre growth. Compared with those of peat, the acidic properties of the local yellow brown soil in the subtropical high-mountain regions were more favorable for S. palustre growth. As N addition increased, the four growth indicators responded inconsistently to the different substrates. While the number of capitula markedly increased, the other three indicators significantly decreased in the mountain soil or exhibited no definitive changes in the peat. The addition of P markedly promoted S. palustre growth in both substrates. However, a threshold for P fertilization existed; the highest productivity occurred at P additions of 0.2 and 0.5 g m^?2 year^?1 in the peat and mountain soil, respectively. The N and P contents in the capitula increased in parallel as the N and P fertilization rates increased, suggesting that these nutrients were absorbed proportionately and were used during the growth of S. palustre.     

Development of the CELLOP optimisation model for plant cell cultivation

A mathematical computer-aided model CELLOP was constructed in which the desirability functions in a three-dimensional experimental design can be used to find the optimal growing conditions for plant cells. CELLOP is intended for the optimisation of 3 to 6 physical, chemical, or biological variables in the cultivation conditions of plant cell cultures. The model was used to optimise the culturing conditions (calcium, inorganic nitrogen, and sucrose concentrations) for coumarin-producing, spontaneously embryogenic cell lines of angelica Angelica archangelica L. subsp. archangelica and hogs fennel Peucedanum palustre (L.) Moench. For A. archangelica the overall optimum concentrations were 0.47 mM Ca²⁺, 5.06 mM NO₃ ⁻, 0.40 mM NH₄ ⁺, and 96.25 mM sucrose. The dry mass was 24.7 % higher and the coumarin content 40.5 % higher than those achieved in the standard 75 % Gamborg B5 medium. For A. archangelica the highest embryogenic activity was reached in the media containing 1.25 mM Ca²⁺. In the case of P. palustre the overall optimum concentrations were 1.60 mM Ca²⁺, 2.84 mM NO³ ₋, 0.23 mM NH₄ ⁺, and 85.10 mM sucrose. For P. palustre the dry mass production increased by 61.8 % and the coumarin content by 58.1 % compared to the values achieved in the Gamborg B5 medium. For P. palustre the highest embryogenic activity was reached in the presence of 50.0 mM NO₃ ⁻ and 4.01 mM NH₄ ⁺.     

Identification of cardiac glycosides in fractions from Periploca forrestii by high-performance liquid chromatography/diode-array detection/electrospray ionization multi-stage tandem mass spectrometry and liquid chromatography/nuclear magnetic resonance

Cardiac glycosides are a class of naturally occurring compounds that are characterized by some interesting biological activities and are widely distributed in the plant kingdom and can also be found in some animals. There is an interest in the chemical characterization of these molecules due to their toxicity and their use in medicines. In the study reported here, a combination of electrospray ionization tandem mass spectrometry with high-performance liquid chromatography equipped with diode-array detector (HPLC-DAD/ESI-MSⁿ), and hyphenation to both liquid chromatography and nuclear magnetic resonance spectroscopy (HPLC/NMR) were utilized for the on-line analyses of cardiac glycosides from Periploca forrestii. The fragmentation patterns and ¹H NMR spectra of nine isolated cardiac glycosides were investigated; their fragmentation rules and ¹H NMR spectral characteristics were summarized and applied to the structural identification of similar constituents in fractions from P. forrestii. As a result, a total of nine trace cardiac glycosides were tentatively determined by analyses of accurate molecular masses, representative fragment ions and characteristic ¹H NMR signals provided by HPLC/high-resolution mass spectrometry (HRMS), HPLC-DAD/ESI-MSⁿ and HPLC/¹H NMR experiments, respectively. Of these, eight (2–9) are new compounds and one (1) is reported from P. forrestii for the first time. Results of the present study can benefit the rapid identification and targeted isolation of new cardiac glycosides from crude plant extracts.     

Profiling of Coumarins in Peucedanum palustre (L.) Moench Populations Growing in Finland

The coumarin composition of Peucedanum palustre (L.) Moench populations growing in Finland was investigated. A total of 132 flowering P. palustre specimens from 43 locations in southern and central Finland were collected, divided into root, stem, leaf, and umbel samples, and analyzed by HPLC. HPLC coupled to high‐resolution mass spectrometry was used to aid the identification of coumarins. A total of 13 coumarin‐structured compounds were quantitatively analyzed from the samples. The coumarin profile of root samples was found to differ from the aerial plant parts. The main coumarins in roots were oxypeucedanin and columbianadin. In aerial parts, peulustrin isomers were the most abundant coumarin components. Umbels and leaves also contained a considerable amount of umbelliprenin, which was only found in traces in roots. Based on hierarchical cluster analysis of the coumarin profiles, some populations shared common characteristics. The most distinct property connecting certain populations was their high peulustrin content. Another notable common property between some populations was the high umbelliprenin content in aerial plant parts. Some populations were clustered together due to their low overall coumarin content.     

Matrix Assisted Ionization Vacuum (MAIV), a New Ionization Method for Biological Materials Analysis Using Mass Spectrometry

The introduction of electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI) for the mass spectrometric analysis of peptides and proteins had a dramatic impact on biological science. We now report that a wide variety of compounds, including peptides, proteins, and protein complexes, are transported directly from a solid-state small molecule matrix to gas-phase ions when placed into the vacuum of a mass spectrometer without the use of high voltage, a laser, or added heat. This ionization process produces ions having charge states similar to ESI, making the method applicable for high performance mass spectrometers designed for atmospheric pressure ionization. We demonstrate highly sensitive ionization using intermediate pressure MALDI and modified ESI sources. This matrix and vacuum assisted soft ionization method is suitable for the direct surface analysis of biological materials, including tissue, via mass spectrometry.The conversion of large and nonvolatile compounds such as proteins into gas-phase ions is of immense fundamental and practical importance. The 2002 Nobel Prize in Chemistry was awarded for the accomplishment of this conversion via electrospray ionization (ESI)¹ (1) and matrix-assisted laser desorption/ionization (MALDI) (2) interfaced with mass spectrometry (MS) to obtain the molecular weights of proteins with high accuracy. These methods employ high voltage or a laser to form gaseous analyte ions from a wide variety of compounds in solution or a solid matrix, respectively.MALDI interfaced with a time-of-flight (TOF) mass spectrometer produces gas-phase analyte ions in vacuum and is the method of choice for the molecular imaging of biological surfaces. Ionization in vacuum provides excellent ion transmission (3), as well as good spatial resolution achieved using a focused laser beam. However, the analysis of protein complexes is very challenging with MALDI, requiring strategies such as first-shot phenomena (4) and chemical crosslinking (5). The necessity of a laser also makes MALDI less soft than ESI and produces background ions, which can hinder the analysis of small molecules (6, 7). MALDI is also of limited utility on high performance mass-to-charge (m/z) analyzers because of mass range issues related to the formation of singly charged ions, which also produce few fragment ions for structural characterization (8).Multiple charged ions produced directly from solution in ESI bring the m/z ratio within the range of high performance mass spectrometers, allowing the analysis of high-mass compounds. These instruments have advanced features for structural characterization, such as ion mobility spectrometry (IMS) for gas-phase separations (9–11), ultra-high mass resolution and mass accuracy (12–14), and advanced fragmentation such as electron transfer dissociation (ETD) (13, 14). However, ESI is limited for surface characterization, requiring approaches such as desorption-ESI (15) and laser ablation ESI (16), ionization methods that produce multiply charged ions but are not compatible with analyses of larger proteins or fragile complexes.A softer ionization approach is needed in order to observe fragile molecules and molecular complexes in living organisms at low levels directly from tissue and cell cultures, without extensive sample preparation, while retaining spatial information. Ideally, this approach would be compatible with mass spectrometers having advanced capabilities to aid structural characterization directly from surfaces. The new ionization method described here, in which molecules are transferred from solid-phase to gas-phase ions through the simple exposure of a material of interest in a suitable matrix to vacuum, is an advance toward this goal and is of fundamental interest.     

Chemical composition,antioxidant and antimicrobial activities and characterization of bioactive compounds from essential oil of <Emphasis Type="Italic">Cinnamomum tamala</Emphasis> grown in north-western Himalaya

The essential oil (EO) obtained from the fresh and dried leaves of Cinnamomum tamala was analysed by gas chromatography/mass spectrometry. EO from fresh leaves showed the presence of 21 compounds, whereas, EO from the dried leaves of C. tamala showed the presence of 20 compounds. In vitro assays namely scavenging ability against 1,1-diphenyl-2-picrylhydrazyl, reducing power and chelating ability on Fe²⁺ ions were used to determine the antioxidant potential of EO of C. tamala. With regard to antifungal activity, EO from dried leaves was more effective against Alternaria alternata and Curvularia lunata than the EO from fresh leaves. Similarly, EO from C. tamala leaves also showed potent antibacterial activity against two Gram negative and two Gram positive bacteria namely, Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus and Pseudomonas aeruginosa. Furthermore, bioactive molecule from C. tamala EO having antifungal and antioxidant activity was isolated and characterized using bioautography, preparative thin layer chromatography and GC/MS analysis and was determined as eugenol. Its minimum inhibitory amount against A. alternata and C. lunata was determined using bioautography assay and was found to be 9.5 and 8.2 µg respectively.     

Influence of the phenolic compound bearing species Ledum palustre on soil N cycling in a boreal hardwood forest

The effects of the understory shrub Ledum palustre on soil N cycling were studied in a hardwood forest of Interior Alaska. This species releases high concentrations of phenolic compounds from green leaves and decomposing litter by rainfall. Organic and mineral soils sampled underneath L. palustre and at nearby non-Ledum sites were amended with L. palustre litter leachates and incubated at controlled conditions. We aimed to know (i) whether L. palustre presence and litter leachate addition changed net N cycling rates in organic and mineral soils, and (ii) what N cycling processes, including gross N mineralization, N immobilization and gross N nitrification, were affected in association with L. palustre. Our results indicate that N transformation rates in the surface organic horizon were not affected by L. palustre presence or leachate addition. However, mineral soils underneath L. palustre as well as soils amended with leachates had significantly higher C/N ratios and microbial respiration rates, and lower net N mineralization and N-to-C mineralization compared to no Ledum and no leachates soils. No nitrification was detected. Plant presence and leachate addition also tended to increase both gross N mineralization and immobilization. These results suggest that soluble C compounds present in L. palustre increased N immobilization in mineral soils when soil biota used them as a C source. Increases in gross N mineralization may have been caused by an enhanced microbial biomass due to C addition. Since both plant presence and leachate addition decreased soil C/N ratio and had similar effects on N transformation rates, our results suggest that litter leachates could be partially responsible for plant presence effects. The lower N availability under L. palustre canopy could exert negative interactions on the establishment and growth of other plant species.     

Bioreactor shoot cultures of <Emphasis Type="Italic">Rhododendron tomentosum</Emphasis> (<Emphasis Type="Italic">Ledum palustre</Emphasis>) for a large-scale production of bioactive volatile compounds

Rhododendron tomentosum Harmaja (Ledum palustre), a peat bog plant from Ericaceae family, has been used in traditional medicine as the anti-arthritis agent. Although modern researches confirm its anti-inflammatory properties, it remains threatened by habitat degradation and possibilities to collect this endangered species from its natural environment for further biological activity studies are limited. Therefore, R. tomentosum liquid in vitro cultures were established as the alternative source of that valuable plant material. Schenk–Hildebrandt medium with 24.60 μM 2-isopentenyladenine and 592.02 μM adenine provides intensive growth and proper morphology of the obtained microshoots. The R. tomentosum biomass was scaled up using the various bioreactors (immersion, temporary immersion and spraying systems) for better growth and improved volatile oil production. The largest biomass accumulation (fresh weight?=?250 g l^?1, growth index?=?280, dry weight?=?20 g l^?1) and essential oil content (0.5% v/m) were achieved with application of commercially available RITA^® bioreactor. GC/MS analysis revealed the high content of p-cymene (6.9%), alloaromadendrene (5.5%), shyobunone (8.2%) and ledene oxide (II) (13.0%) in the volatile fraction obtained from RITA^® system. The biomass growth parameters and production profile in terms of essential oil and selected terpenoid compounds were determined during the 2 month period. The influence of culture conditions and bioreactor construction on the growth and volatile oil production in R. tomentosum biomasses was discussed.     

The influence of goethite and gibbsite on soluble nutrient dynamics and microbial community composition

Iron and aluminum (oxyhydr)oxides are ubiquitous in the soil environment and have the potential to strongly affect the properties of dissolved organic matter. We examined the effect of oxide surfaces on soluble nutrient dynamics and microbial community composition using an incubation of forest floor material in the presence of (1) goethite and quartz, (2) gibbsite and quartz, and (3) quartz surfaces. Forest floor material was incubated over a period of 154 days. Aqueous extracts of the incubations were harvested on days 5, 10, 20, 30, 60, 90, and 154, and concentrations of P, N, PO₄ ^3?, NO₂ ^?, NO₃ ^?, and organic C were measured in the solutions. Microbial community composition was examined through pyrosequencing of bacterial and fungal small subunit ribosomal RNA genes on selected dates throughout the incubation. Results indicated that oxide surfaces exerted strong control on soluble nutrient dynamics and on the composition of the decomposer microbial community, while possibly having a small impact on system-level respiration. Goethite and gibbsite surfaces showed preferential adsorption of P-containing and high molar mass organic solutes, but not of N-containing compounds. On average, organic C concentrations were significantly lower in water extractable organic matter (WEOM) solutions from oxide treatments than from the control treatment (P = 0.0037). Microbial community composition varied both among treatments and with increasing time of incubation. Variation in bacterial and fungal community composition exhibited strong-to-moderate correlation with length of incubation, and several WEOM physiochemical characteristics including apparent (weight averaged) molar mass, pH and electrical conductivity. Additionally, variation in bacterial community composition among treatments was correlated with total P (r = 0.60, P < 0.0001), PO₄ ^3? (r = 0.79, P < 0.0001), and organic C (r = 0.36, P = 0.015) concentrations; while variation in fungal communities was correlated with organic C concentrations (r = ?0.48, P = 0.0008) but not with phosphorus concentrations. The relatively small impact of oxide surfaces on system-level microbial respiration of organic matter despite their significant effects on microbial community composition and WEOM dynamics lends additional support to the theory of microbial functional redundancy.     

Diversity and Antagonistic Potential of Bacteria Associated with Bryophytes from Nutrient-Poor Habitats of the Baltic Sea Coast

Very little is known about the interaction of bryophytes with bacteria. Therefore, we analyzed bacteria associated with three bryophyte species, Tortula ruralis, Aulacomnium palustre, and Sphagnum rubellum, which represent typical moss species of three nutrient-poor plant communities at the southern Baltic Sea coast in Germany. By use of two cultivation-independent techniques, denaturing gradient gel electrophoresis and single-strand conformation polymorphism analysis of the 16S ribosomal DNA, a high degree of moss specificity was found for associated bacterial communities. This specificity could be further evidenced by a cultivation-dependent approach for the following parameters: (i) plate counts of bacteria on R2A medium, (ii) proportion of antagonistic isolates, (iii) antagonistic activity as well as spectrum against pathogens, and (iv) diversity and richness of antagonistic isolates. The proportion of isolates with antagonistic activity against the pathogenic model fungus Verticillium dahliae was highest for S. rubellum (31%), followed by A. palustre (17%) and T. ruralis (5%). A high percentage (99%) of moss-associated antagonistic bacteria produced antifungal compounds. The high recovery of antagonistic isolates strongly suggests that bryophytes represent an ecological niche which harbors a diverse and hitherto largely uncharacterized microbial population with yet unknown and untapped potential biotechnological applications, e.g., for biological control of plant pathogens.     

Microbial Colonization and Competition on the Marine Alga Ulva australis

Pseudalteromonas tunicata and Roseobacter gallaeciensis are biofilm-forming marine bacteria that are often found in association with the surface of the green alga Ulva australis. They are thought to benefit the plant host by producing inhibitory compounds that are active against common fouling organisms. We investigated factors that influence the ability of P. tunicata and R. gallaeciensis to attach to and colonize the plant surface and also the competitive interactions that occur between these organisms and other isolates from U. australis during biofilm formation on the plant surface. A surprisingly high number of P. tunicata cells, at least 10⁸ cells ml⁻¹, were required for colonization and establishment of a population of cells that persists on axenic surfaces of U. australis. Factors that enhanced colonization of P. tunicata included inoculation in the dark and pregrowth of inocula in medium containing cellobiose as the sole carbon source (cellulose is a major surface polymer of U. australis). It was also found that P. tunicata requires the presence of a mixed microbial community to colonize effectively. In contrast, R. gallaeciensis effectively colonized the plant surface under all conditions tested. Studies of competitive interactions on the plant surface revealed that P. tunicata was numerically dominant compared with all other bacterial isolates tested (except R. gallaeciensis), and this dominance was linked to production of the antibacterial protein AlpP. Generally, P. tunicata was able to coexist with competing strains, and each strain existed as microcolonies in spatially segregated regions of the plant. R. gallaeciensis was numerically dominant compared with all strains tested and was able to invade and disperse preestablished biofilms. This study highlighted the fact that microbial colonization of U. australis surfaces is a dynamic process and demonstrated the differences in colonization strategies exhibited by the epiphytic bacteria P. tunicata and R. gallaeciensis.     

Isolation of polyketides from Prymnesium parvum (Haptophyta) and their detection by liquid chromatography/mass spectrometry metabolic fingerprint analysis

Prymnesium parvum is a microalga that forms blooms coupled with the presence of potent exotoxins; however, no chemical standards are currently available for the toxins. Streamlined methods are presented for the separation and enrichment of polyketide toxins, prymnesin-1 (prym1) and prymnesin-2 (prym2). Prymnesins were separated by reversed-phase chromatography and detected by positive-mode electrospray ionization MS to generate a unique metabolic fingerprint. More than 10 ions were detected and mass assignments were in agreement with predicted isotopic distributions for the intact compounds and related fragments; ions occurred as multiply protonated species and with common salt adducts. The most prevalent ion was observed at 919.88 m/z, which represents the aglycone [prym_agly + 2H]²⁺ backbone structure common to both molecules. Expanded mass spectra for this and related ions were in excellent agreement (<0.5 ppm) with empirically derived spectra based on elemental composition and naturally occurring isotopes. These investigations have confirmed the isolation of polyketide prymnesins from whole cells, which heretofore has not been reproduced since their original characterization. Moreover, this study represents the first time these compounds have been verified in aqueous materials. These tools should allow the direct identification and analysis of polyketide prymnesins, which will greatly improve our understanding of these toxins in P. parvum.     

The potential application of FT-Raman spectroscopy for the quantification and mapping of the steroidal glycoside P57 in Hoodia gordonii

Hoodia gordonii, with the perceived active ingredient P57 (a steroidal glycoside), is a succulent plant which has gained commercial popularity as an anti-obesity preparation. The content of P57 is used as an indication of the quality of the raw material. Traditionally, quantification of P57 is performed using liquid-chromatography coupled to mass spectrometry (LC–MS) which is expensive and laborious. Vibrational spectroscopy methods such as FT-Raman spectroscopy offer a simple, less expensive and rapid alternative. The potential of FT-Raman to quantify and identify the location of P57 in H. gordonii raw plant material was investigated. LC–MS was used to determine the concentration of P57 in 145 plant samples and the data was used to develop a calibration model with chemometric techniques based on the partial least squares projections to latent structures (PLS) algorithm. The performance of the calibration model was evaluated according to the root mean square error of prediction (RMSEP) and correlation coefficient (R²). Pre-processing with orthogonal signal correction (OSC) yielded a model which predicted P57 content based on the FT-Raman spectra with a correlation coefficient (R²) value of 0.9986 and an RMSEP of 0.004%. These results demonstrate that FT-Raman spectroscopy holds great potential to rapidly quantify P57 in H. gordonii raw material with high accuracy as an alternative to LC–MS analysis. In addition, the spatial distribution of P57 in a cross-section of an H. gordonii stem sample was demonstrated using FT-Raman mapping showing that P57 is concentrated throughout the cortex which was confirmed with LC–MS.     

Chemo-profiling of bioactive metabolites from Chaetomium globosum for biocontrol of Sclerotinia rot and plant growth promotion

Chaetomium globosum Kunze ex. Fries has been known to produce diverse bioactive metabolites, attracting researchers to exploit the biocontrol agent for plant disease management. However, distinct research gaps are visible regarding detail characterization of bioactive metabolites. Thus the current study has been planned to characterize volatile and nonvolatile compounds of most potential strain of C. globosum 5157. GC–MS analysis of hexane fraction revealed twenty-six volatile organic compounds, representing 65.5% of total components in which 3-octanone (21.4%) was found to be most abundant. UPLC-QTOF-MS/MS analysis of ethyl acetate and methanolic fractions resulted tentative characterization of fifteen and eleven metabolites, respectively. Among these, nine metabolites were isolated, purified and characterized using ¹H NMR and High resolution mass spectrometric analysis to delineate mass fragmentation pattern for the first time. Antifungal potential of hexane fraction exhibited high inhibitory action against Sclerotium rolfsii (139.2 μg mL^?1) whereas ethyl acetate fraction was highly effective against Sclerotinia sclerotiorum (112.1 μg mL^?1). Comparative assessment of C. globosum 5157 vis a vis Trichoderma harzianum A28 revealed promising effect of C. globosum 5157 with respect to antifungal properties and plant growth promotion of Brassica seedlings.     

Quantification of plant surface metabolites by matrix‐assisted laser desorption–ionization mass spectrometry imaging: glucosinolates on Arabidopsis thaliana leaves

The localization of metabolites on plant surfaces has been problematic because of the limitations of current methodologies. Attempts to localize glucosinolates, the sulfur‐rich defense compounds of the order Brassicales, on leaf surfaces have given many contradictory results depending on the method employed. Here we developed a matrix‐assisted laser desorption–ionization (MALDI) mass spectrometry protocol to detect surface glucosinolates on Arabidopsis thaliana leaves by applying the MALDI matrix through sublimation. Quantification was accomplished by spotting glucosinolate standards directly on the leaf surface. The A. thaliana leaf surface was found to contain approximately 15 nmol of total glucosinolate per leaf with about 50 pmol mm^?2 on abaxial (bottom) surfaces and 15–30 times less on adaxial (top) surfaces. Of the major compounds detected, 4‐methylsulfinylbutylglucosinolate, indol‐3‐ylmethylglucosinolate, and 8‐methylsulfinyloctylglucosinolate were also major components of the leaf interior, but the second most abundant glucosinolate on the surface, 4‐methylthiobutylglucosinolate, was only a trace component of the interior. Distribution on the surface was relatively uniform in contrast to the interior, where glucosinolates were distributed more abundantly in the midrib and periphery than the rest of the leaf. These results were confirmed by two other mass spectrometry‐based techniques, laser ablation electrospray ionization and liquid extraction surface analysis. The concentrations of glucosinolates on A. thaliana leaf surfaces were found to be sufficient to attract the specialist feeding lepidopterans Plutella xylostella and Pieris rapae for oviposition. The methods employed here should be easily applied to other plant species and metabolites.     

Structural characterization of trace stilbene glycosides in Lysidice brevicalyx Wei using liquid chromatography/diode-array detection/electrospray ionization tandem mass spectrometry

The mass fragmentation patterns of stilbene glycosides isolated from the genus Lysidice were investigated by negative ion electrospray ionization tandem mass spectrometry, and the influence of collision energy on their fragmentation behavior is discussed. It is found that the presence of the Y₀⁻ and B₀⁻ ions in the MS² spectra is characteristic for 1 → 6 linked diglycosyl stilbenes, while the Y₀⁻, Y₁⁻, and Z₁⁻ ions are representative ions of 1 → 2 linked diglycosyl stilbenes. These results indicate that ESI-MSⁿ in the negative ion mode can be used to differentiate 1 → 6 and 1 → 2 linked diglycosyl stilbenes. Based on the fragmentation rules, 9 new trace constituents were identified or tentatively characterized in a fraction of Lysidice brevicalyx by using HPLC/HRMS and HPLC-DAD/ESI-MSⁿ. The results of the present study can assist in on-line structural identification of analogous constituents and targeted isolation of novel compounds from crude plant extracts.     

Chemical Composition of Lipids from Peat-Forming Leafy Mosses of Eutrophic Swamps of Western Siberia and Altai

The molecular composition of lipids in three samples of leafy mosses (Aulacomnium palustre, Warnstorfia fluitans, and Calliergon giganteum) has been determined. The revealed acyclic compounds included normal and isoprenoid alkanes, isoprenoid alkenes, normal and isoprenoid ketones, carboxylic acids and their esters, alcohols, and aldehydes. Among cyclic compounds, bi-, tri- and tetracyclic polycycloaromatic hydrocarbons (PAHs), bicyclic and pentacyclic terpenoids, steroids and tocopherols have been observed. The identified organic compounds consisted mainly of carbocyclic acids and n-alkanes with the prevalence of C₂₇ homologues. A. palustre is characterized by a reduced content of isoprenoid compounds, alcohols, and ketones, while the content of unsaturated acids, pentacyclic terpenoids, and aldehydes is rather heightened. A. palustre differs from W. fluitans and C. giganteum in the steroid composition and contains eremophylene, a sesquiterpenoid, which is absent in the mosses of the family Amblystegiaceae. Compared to C. giganteum, W. fluitans has a higher content of lycopadiene, carboxylic acids, n-alkanes, phyt-2-ene, aldehydes, esters, squalene, diploptene, α-tocopherol, and triphenyl phosphates.     

Determination of naphthodianthrones in plant extracts from Hypericum perforatum L. by liquid chromatography–electrospray mass spectrometry

The determination of the naphthodianthrone constituents in extracts of dried blossoms of Hypericum perforatum L. by combined HPLC–electrospray mass spectrometry is described. Hypericin (1), pseudohypericin (2) and their precursor compounds produce intensive negative quasi-molecular ions by deprotonation provided a non-acidic eluent system is used in the HPLC separation. From the [M–H]⁻ ions formed in the electrospray ionization process characteristic daughter ion spectra can be obtained by collisional activation which have been studied by tandem mass spectrometry.     

Identification of maloyl glucans from Euphorbia tirucalli by ESI-(−)-FT-ICR MS analyses

Euphorbia tirucalli aerial parts are popularly used in Brazil for cancer treatment. The elution of the aqueous extract of the plant on silica gel C-18 cartridge furnished a water-soluble fraction, which was analyzed directly into the electrospray ionization (ESI) source combined with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) and characterized as a mixture of malic acid glycosides 1–5. The compounds were detected in their deprotonated form [M−H]⁻, where their exact mass (mass error lower than 1 ppm), molecular formula (C_nH_hO_o), double bond equivalent (DBE) and connectivity were determined from ESI-(−)-MS and ESI-(−)-MS/MS experiments. The presence of malic acid and glucose, as part of the structures, could originate from crassulacean acid metabolism (CAM) of the plant.     

Phosphatase and phytase activities in nodules of common bean genotypes at different levels of phosphorus supply

Plants grown at limited P supply can increase the activity of phosphatases in roots to hydrolyse organic-P compounds in the soil thus improving plant P acquisition, but little information is available about the role of these enzymes for internal plant metabolism at limited-P conditions. This work intended to measure the activities of acid phosphatases and phytases in nodules of common bean (Phaseolus vulgaris) genotypes at different levels of P supply. The experiment was carried out in a 5?×?5 factorial design with four replicates, comprising five bean genotypes and five P levels (20, 40, 80, 160 and 320 μmol P plant^?1 week^?1) in nutrient solution. Root seedlings were inoculated with Rhizobium tropici and plants were grown in 1-l bottles. Nodule samples were detached from 39-day-old plants and enzyme activities were determined in crude extracts. Plants were harvested at the stage of pod setting. Polynomial models fitted to data indicated maximal values at the level of 194 μmol P for shoot mass, at 206 μmol P for nodule mass and at 221 μmol P for shoot N. Whereas shoot mass was 1.7 times lower at 20 than at 160 μmol P, nodule mass was 7.5 times lower. Concentration of P in nodules increased from 40 to 320 μmol P but remained stable between 20 and 40 μmol P, suggesting a minimal threshold concentration of 3 mg P g^?1 for nodule growth. Activities of phosphatases and phytases in nodules decreased strongly as P supply was raised from 20 to 80 μmol P, remaining almost stable at higher P levels. Phosphatase activity ranged from 1,154 to 406 nmol min^?1 g^?1 (nodule fresh mass) from 20 to 80 μmol P respectively, while the phytase activity ranged from 55 to 14 nmol min^?1 g^?1 from 20 to 80 μmol P. Bean genotypes differed in shoot and nodule mass at the levels of 80 and 160 μmol P, whilst they differed in nodule enzyme activities only at the lowest P level, the relationship between nodule enzyme activities and growth of different bean genotypes was not evident. It is concluded that bean plants at P-deficient conditions increase the activities of phosphatases and phytases in nodules. This may constitute an adaptive mechanism for N₂-fixing legumes to tolerate P deficiency, by increasing the utilisation of the scarce P within the nodules.