Occurrence of rosmarinic acid,chlorogenic acid and rutin in Marantaceae species

In a survey of the higher plants for families with rosmarinic acid-accumulating species we could show for the first time, that some species of the family Marantaceae of the order Zingiberales accumulate rosmarinic acid. Other compounds detected in Marantaceae are chlorogenic acid and rutin (quercetin 3-O-rutinoside). Out of 35 species coming from 9 different genera extracted and analysed, two species of Maranta (Maranta leuconeura, Maranta depressa) and one Thalia species (Thalia geniculata) showed the presence of rosmarinic acid. The two Maranta species additionally contained chlorogenic acid, which was also present in Stromanthe amabilis. Rutin was detected in the genera Calathea, Ctenanthe, Maranta, Pleiostachya and Thalia. For a comparison, species from six other families of the Zingiberales were analysed as well.     

Caffeic acid ester derivatives and flavonoids of genus Arnaldoa (Asteraceae,Barnadesioideae)

The phytochemical composition of Arnaldoa species is barely known. In this work, the occurrence of caffeic acid ester derivatives and flavonoids in A. argentea, A. macbrideana and A. weberbaueri was established by liquid chromatography associated to high-resolution mass spectrometry analyses and comparison with data from isolated compounds. The distribution of chlorogenic acids in the genus Arnaldoa is herein described for the first time. The metabolite profile of Arnaldoa species was compared to that of Tithonia diversifolia, a known and rich source of chlorogenic acids and sesquiterpene lactones. In addition to the mono- and dicaffeoyl quinic acids present in T. diversifolia, Arnaldoa species exhibited the mono- and dicaffeoyl tartaric acids. Furthermore, mass features correspondent to that of sesquiterpene lactones present in T. diversifolia were not observed in Arnaldoa species. The chemotaxonomic implications of caffeic acid ester derivatives and flavonoid glycosides, as well as the potential absence of sesquiterpene lactones in the genus Arnaldoa and subfamily Barnadesioideae are discussed.     

Phenolic acid content,antioxidant and cytotoxic activities of four Kalanchoë species

Phenolic acid composition, antioxidant, and cytotoxic activities in leaves of four Kalanchoe (Crassulaceae) species were evaluated. Determination of phenolic acid contents were conducted by an optimized LC–ESI-MS/MS method. The results show that Kalanchoe daigremontiana Raym.-Hamet & H. Perrier (using ASE extraction) and Kalanchoe pinnata (Lam.) Pers. contain the highest amounts of phenolic acids, while Kalanchoe nyikae Engl. the lowest ones. Among phenolic acids ferulic, caffeic and protocatechuic acids were occurring in the highest quantities in the analysed species. The greatest amounts of ferulic and protocatechuic acids were found in K. daigremontiana and K. pinnata. Moreover, the antiradical and cytotoxic activities of Kalanchoe extracts were investigated. All tested extracts possessed antioxidant activity. The obtained IC₅₀ values (μg/mL) ranged from 49.9 μg/mL to 1410 μg/mL, indicating a large variation of the activity of the analysed extracts. Cytotoxicity assays revealed dose-dependent effects in the cells lines tested. Only K. pinnata extract showed a high cytotoxicity against the H-9 human T cell line. Other extracts (K. daigremontiana, Kalanchoe milloti, K. nyikae) showed more pronounced cytotoxicity towards J45.01 cells (human acute lymphoblastic leukaemia T cells).The present study demonstrated that Kalanchoe extracts have significant antioxidant and cytotoxic effects. This suggests that these species can be used as new sources of natural antioxidants and potential anticancer compounds.     

ThenadB gene ofSalmonella typhimurium complements the nicotinic acid auxotrophy ofShigella flexneri

Shigella species are characteristically nicotinic acid (NA) auxotrophs. The invasiveS. flexneri strain M90T, transformed with the multicopy plasmid pZT349 encoding thenadB gene ofSalmonella typhimurium, can grow in minimal glucose medium without exogenous NA, whereas, M90T containing the control vector, pUC18 does not, suggesting that this species lacksl-aspartic acid oxidase, the first enzyme in the de novo NAD biosynthetic pathway. The estimated growth rate of strain M90T (pZT349) in HeLa cells was identical to that of M90T (pUC18), indicating the available intracellular concentration of NA is not limiting for bacterial growth.     

Lipid, fatty acid and protein utilization during lecithotrophic larval development of Lithodes santolla (Molina) and Paralomis granulosa (Jacquinot)

During the larval development of the subantarctic king crab, Lithodes santolla, and stone crab, Paralomis granulosa, we compared changes in the carbon, fatty acid and protein contents of larvae reared under constant conditions from hatching to metamorphosis, either in presence or absence of food (Artemia spp. nauplii). In both species the feeding condition had no influence on any of the chemical parameters studied, indicating a fully lecithotrophic (i.e. non-feeding) mode of development from hatching of the first zoea to metamorphosis of the late megalopa. Dry mass and carbon contents at hatching were similar in the larvae of both species, but L. santolla contained initially higher total amounts of fatty acids and protein than P. granulosa. Both species utilized considerable portions of their total fatty acid pool which decreased logarithmically throughout the time of development. At metamorphosis, it was almost exhausted in P. granulosa, while L. santolla had consumed only about 60%. Protein utilization, in contrast, was higher in L. santolla (40%) than in P. granulosa (20%). Triacylglycerol was the principal storage lipid in both species, accounting initially for about 75% of the lipid fraction; it was strongly utilized during larval development. Phospholipid constituted the second largest lipid class; it also decreased in P. granulosa, but to a lesser extent in L. santolla. The major fatty acids of both species were 18:1(n−9), 20:5(n−3) and 16:0 as well as, in lower proportions, 18:1(n−7), 22:6(n−3), 16:1(n−7) and 18:0. Monounsaturated fatty acids represented the dominant group in L. santolla, whereas P. granulosa contained similar amounts of mono- and polyunsaturated fatty acids. In L. santolla, monounsaturated fatty acids, especially 16:1(n−7), were preferentially utilized as compared to polyunsaturates. Due to a particularly strong lipid utilization in P. granulosa, all individual fatty acids were largely depleted at metamorphosis, showing similar extents of consumption. L. santolla had higher initial lipid and protein stores that seem to be used more economically as compared to P. granulosa.     

Sterols and fatty acids of some non-photosynthetic angiosperms

Sterols and fatty acids were extracted and identified from three parasitic angiosperms, Cuscuta campestris, Monotropa uniflora and M. hypopitys. Each plant contained the typical 16 and 18-carbon fatty acids of angiosperms, but the partially-photosynthetic Cuscuta contained much larger quantities of linolenic acid than the non-green Monotropa species which had smaller amounts of linolenic acid characteristic of non-photosynthetic tissue. Sterol quantity was three times higher in Cuscuta than in the Monotropa species. Sitosterol was the major sterol in all species with smaller amounts of campesterol and cholesterol.     

Sialic acid attenuates the cytotoxicity of the lipid hydroperoxides HpODE and HpETE

Reduction of peroxide molecular species is an essential function in living organisms. In previous studies, we proposed a new function for the sialic acid N-acetylneuraminic acid (Neu5Ac)—that of antioxidant/hydrogen peroxide scavenging agent. On the basis of the reaction scheme, Neu5Ac is thought to act as a general antioxidant of all hydroperoxide-type species (R-OOHs). The concentration of tert-butyl hydroperoxide (t-BuOOH) decreased after co-incubation with N-acetylneuraminic acid. Neu5Ac also decreased the R-OOH concentration in solutions of peroxylinolenic acid (13(S)-hydroperoxy-(9Z,11E)-octadecadienoic acid, HpODE) and peroxyarachidonic acid (15(S)-hydroperoxy-(5Z,8Z,11Z,13E)-eicosatetraenoic acid, HpETE)—two lipid hydroperoxides that participate in many physiological events. Moreover, the cytotoxicity of both these lipid hydroperoxides was attenuated by reaction with Neu5Ac acid. Our results suggest that N-acetylneuraminic acid is a potential antioxidant of most hydroperoxides that accumulate in organisms.     

Expression of amino acid transporter genes in developmental stages and adult tissues of Antarctic echinoderms

Epithelial cells in the body wall of adult and developmental stages of marine invertebrates absorb dissolved organic material directly from seawater. Despite over a century of study, little is known about the molecular biological mechanisms responsible for this transport process. Previous studies on embryonic and larval Antarctic echinoderms show that amino acid uptake could provide an important supplement of metabolic substrates. In the present study, partial cDNA sequences of 11 putative amino acid transporter genes were isolated from six species of Antarctic echinoderms including the Antarctic sea stars Acodontaster hodgsoni, Diplasterias brucei, Odontaster meridionalis, Odontaster validus, and Perknaster fuscus, and the Antarctic sea urchin Sterechinus neumayeri. Conserved domains of cDNA-deduced amino acid sequences characterized these genes as being members of a family of amino acid transporters (solute carrier family 6). Expression of these genes was detected throughout embryonic and larval development of two species that have contrasting developmental modes (A. hodgsoni: lecithotrophic; O. meridionalis: planktotrophic). In all six species studied, the expression of amino acid transporter genes was detected in tube feet and digestive organs of adult animals, demonstrating that members of a single amino acid transporter gene family are expressed during the entire life history of a marine invertebrate. The identification of these genes is an important step toward developing a mechanistic understanding of amino acid transport capacities in Antarctic marine invertebrates.     

Fatty acid composition of selected prosthecate bacteria

The cellular fatty acid composition of 14 strains of Caulobacter species and types, two species of Prosthecomicrobium, and two species of Asticcacaulis was determined by gas-liquid chromatography. In most of these bacteria, the major fatty acids were octadecenoic acid (C18:1), hexadecenoic acid (C16:1) and hexadecanoic acid (C16:0). Some cyclopropane and branched chain fatty acids were detected in addition to the straight chained acids. Hydroxytetradecanoic acid was an important component of P. enhydrum but significant amounts of hydroxy acids were not detected in other prosthecate bacteria examined.Abbreviations DEGS diethylene glycol succinate - A measare of association Dedicated to Prof. R. Y. Stanier on the occasion of his 60th birthday     

Evaluation of fatty acid composition among selected amaranth grains grown in two consecutive years

The oil of amaranth grain (Amaranthus spp.) is a rich source of poly-unsaturated fatty acids. In this study, we tested 10 amaranth samples representing two species (Amaranthus cruentus and Amaranthus hypochondriacus) in two consecutive years (2010, 2011). Grain oils were analysed by gas chromatography for their fatty acids profile. In 2010, oil content ranged from 6.4–8.2% for A. cruentus and 6.3–7.9% for A. hypochondriacus. In 2011, the level was 7.1–8.2% and 6.6–8.7% for A. cruentus and A. hypochondriacus, respectively. Linoleic, palmitic, and oleic acids were dominant fatty acids in all of the oil samples. The essential linoleic acid level was 33.3–38.7% (A. cruentus) and 31.7–47.5% (A. hypochondriacus) in 2010 and 34.6–39.9% (A. cruentus) and 34–44.5% (A. hypochondriacus) in 2011. The minority fatty acids, i.e. stearic, α-linolenic, and arachidic acids were also observed. Eicosenoic and behenic acids were present in the grain in trace amounts. Statistical evaluation showed a significant effect of year and species of amaranth on the levels of certain fatty acids. There was a strong positive correlation between oil content and oleic acid, and a negative correlation between oleic acid and either of the other two fatty acids, linoleic and α-linolenic ones.     

Conversion of (2Z,4E)-5-(1′,2′-epoxy-2′,6′,6′-trimethylcyclohexyl)-3-methyl-2,4-pentadienoic acid to xanthoxin acid by Cercospora cruenta, a fungus producing (+)-abscisic acid

(±)-(2Z,4E)-5-(1′,2′-epoxy-2′,6′,6′-trimethylcyclohexyl)-3-methyl-2,4-pentadienoic acid was metabolized by Cercospora cruenta, which has the ability to produce (+)-abscisic acid (ABA), to give (±)-(2Z,4E)-xanthoxin acid, (±)-(2Z,4E)-5′-hydroxy-1′,2′-epoxy-1′,2′-dihydro-β-ionylideneacetic acid, (±)-1′,2′-epoxy-1′,2′-dihydro-β-ionone and trace amounts of ABA.     

Pyridine salvage and nicotinic acid conjugate synthesis in leaves of mangrove species

The metabolic fate of [carbonyl-¹⁴C]nicotinamide was surveyed in leaf disks of seven mangrove species, Bruguiera gymnorrhiza, Rhizophora stylosa, Kandeliaobovata, Sonneratia alba, Pemphis acidula, Lumnitzera racemosa and Avicennia marina, with and without 250 mM NaCl. Uptake of [¹⁴C]nicotinamide by leaf disks was stimulated by 250 mM NaCl in K. candel, R. stylosa, A. marina and L. racemosa. [Carbonyl-¹⁴C]nicotinamide was converted to nicotinic acid and was utilised for the synthesis of nucleotides and nicotinic acid conjugates. Formation of nicotinic acid by the deaminase reaction was rapid; there was little accumulation of nicotinamide in the disks 3 h after administration. Radioactivity from [carbonyl-¹⁴C]nicotinamide was incorporated into pyridine nucleotides (mainly NAD and NADP) in all mangrove leaves, and the rates varied from 2% (in L. racemosa) to 15% (S. alba) of the total radioactivity taken up. NaCl generally reduced nicotinic acid salvage for NAD and NADP. In all mangrove leaf disks, the most heavily labelled compounds (up to 70% of total radioactivity) were trigonelline (N-methylnicotinic acid) and/or nicotinic acid N-glucoside. Trigonelline was formed in all mangrove plants, but N-glucoside synthesis was found only in leaves of A. marina and K. obovata. In A. marina, incorporation of radioactivity into N-glucoside (51%) was much greater than incorporation into trigonelline (2%). In general, NaCl stimulates the synthesis of these pyridine conjugates. The rate of decarboxylation of nicotinic acid in roots of A. marina seedlings was much greater than for the corresponding reaction observed in leaves.     

The distribution of geraniin and mallotusinic acid in the order geraniales

The distribution of geraniin and mallotusinic acid in Geraniaceae, Euphorbiaceae and other families in the Geraniales were investigated by HPLC. Geraniin composes the main part of the tannin in all of the investigated species of Geranium, but not in species of Pelargonium. The species of Geraniaceae lack mallotusinic acid. Geraniin was also detected in most of the species in the subfamily Euphorbioideae of Euphorbiaceae, although the amount was generally smaller than in Geranium, except for some woody species. Several species of Euphorbioideae contained mallotusinic acid. Characteristic of the subfamily Phyllanthoideae is the absence of mallotusinic acid and poor distribution of geraniin. Geraniin and mallotusinic acid were not detected in most of the species of other families in the Geraniales, except in Erythroxylum coca. Hyperin was found to be the main flavonol glycoside in most species of Geranium.     

Antimycobacterial activity of ellagitannin and ellagic acid derivate rich crude extracts and fractions of five selected species of Terminalia used for treatment of infectious diseases in African traditional medicine

Ten crude extracts and their solvent partition fractions from five species of Terminalia collected in Tanzania were assessed for antimycobacterial effects using Mycobacterium smegmatis ATCC 14468 as a model organism. We report here, for the first time, on antimycobacterial effects of root and stem bark extracts of Terminalia sambesiaca and Terminalia kaiserana as well as of fruit extracts of Terminalia stenostachya and leaf extracts of Terminalia spinosa. T. sambesiaca gave the best effects of all the investigated species in terms of the sizes of the inhibitory zones of root and stem bark extracts. A crude methanol root extract of T. sambesiaca gave lower MIC values (1250 μg/ml) than its aqueous and butanol soluble fractions (MIC 2500 μg/ml). Our preliminary HPLC–DAD data indicates that methanol and aqueous extracts of T. sambesiaca roots are rich in ellagitannins and ellagic acid glycosides. Particularly, one polar ellagitannin at t_R 10.3–10.9 min dominates the extracts quantitatively and thus may be responsible for their good antimycobacterial effects. In contrast to the more polar fractions, a chloroform soluble fraction of the roots of T. sambesiaca was devoid of antimycobacterial activity. Also crude methanol and aqueous extracts of the stem bark of T. sambesiaca gave promising antimycobacterial effects (MIC 1250 μg/ml). All fractions of T. kaiserana roots, except from the aqueous insoluble gave good antimycobacterial effects (MIC 1250 μg/ml) and the aqueous extract showed the best effects of the fractions in terms of the size of inhibition zones. These results justify the uses of hot water decoctions of the roots of T. kaiserana for treatment of cough, one of the symptoms of TB. According to HPLC–DAD data methanol extracts of T. kaiserana roots and their aqueous fractions are rich in polar ellagitannins and ellagic acid glycosides. Quantitatively, the ellagitannins dominate these extracts and therefore the good antimycobacterial activity of the methanol and aqueous extracts is assumed to be due to these compounds. Sephadex LH-20 CC fractions of a methanol extract of the roots of T. kaiserana inhibited the growth of M. smegmatis, giving MIC values of 1000 μg/ml. Ellagic acid glycosides in these fractions must be responsible for their good antimycobacterial effects since they are present in high concentrations. Good antimycobacterial effects were also obtained with a root extract of Terminalia sericea, and especially the butanol soluble fraction was a good inhibitor of the growth of M. smegmatis (MIC 1562 μg/ml). Our preliminary HPLC–DAD results show that the roots of T. sericea are rich in ellagitannins, ellagic acid glycosides and at least one stilbene compound. Extracts of the fruits of T. stenostachya gave good antimycobacterial effects, butanol extracts being the most active. Also the leaves of T. stenostachya, and especially the butanol soluble extracts, give good antimycobacterial effects. Our HPLC–DAD data indicate that T. stenostachya leaves contain large quantities of gallic acid, ellagitannins and ellagic acid glycosides. Our results indicate that many of the investigated species of Terminalia might contain leads for development of anti-TB drugs. Standardized extracts of T. sambesiaca, T. kaiserana and T. sericea roots could be used as easily available and cheap medicines for treatment of TB in remote regions of East and South Africa.     

Floral morphology and systematics of the genus Thottea Rottb. (Aristolochiaceae) from the Western Ghats, India

The medicinal genus Thottea (Aristolochiaceae) is represented in the Western Ghats, India, by eight species, seven of which are endemic to this region. In the present study, diversity in floral structure and organization of these eight species was analyzed from a systematic perspective. Floral morphology was thoroughly studied and discussed with emphasis on 16 qualitative and 11 quantitative characters. Statistical tools such as UPGMA cluster analysis, PCoA, PCA, and one-way ANOVA were used for elucidating species boundaries and inter-relationships. Significance of the selected qualitative and quantitative characters for species level systematics was revealed by the PCA and one-way ANOVA. Based on the nature of sepals (free/fused), two species groups were identified (first group with T. duchartrei and T. idukkiana, and the second group with the remaining six species). By considering the number and arrangement of stamens, yet another species grouping was proposed (one with T. abrahamii and T. dinghoui, and the other with the remaining six species). The UPGMA phenogram and the PCoA scatter plot clearly pointed out the existence of two species groups: the first group with T. abrahamii, T. barberi, T. dinghoui, T. ponmudiana, T. siliquosa, and T. sivarajanii, and the second one with T. duchartrei and T. idukkiana. With regard to the morphometric tools and the taxonomic key, it does not seem possible to distinguish T. abrahamii from T. dinghoui, and T. idukkiana from T. duchartrei.     

Phospholipid and acid composition of Nocardia and nocardoid bacteria as criteria of classification

Phospholipid and acid composition of 5 strains of ‘true’ Nocardia and 4 strains of nocardoid bacteria have been studied. A great homogeneity was found in all the Nocardia species: phospholipids consist of cardiolipin, phosphatidyl ethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. Streptomyces (Nocardia) mediterranei did not contain phosphatidylinositol and Oerskovia (Nocardia) turbata had no phosphatidyl ethanolamine. The fatty acid composition of these phospholipids was determined and was found different in Nocardia and nocardoid species. Nocardia were rich in straight chain fatty acids and tuberculostearic acid while the phospholipids of nocardoid bacteria contained greater amounts of branched fatty acids. The fatty acids from acetone soluble lipids consisted of hydroxy and non-hydroxy compounds. Hydroxy acids were found in Nocardia which contained nocardic acids: high MW β-hydroxy α-branched acids and in S. mediterranei which contained β-hydroxy acids with 15–17 carbon atoms. Non-hydroxy acids were essentially palmitic and tuberculostearic acids in Nocardia species while S. mediterranei and O. turbata contained great amounts of iso acids from C₁₄ to C₁₇. Phospholipid and acid composition are discussed as criteria of taxonomic classification of Nocardia and related Actinomycetes.     

Microsymbionts of Phaseolus vulgaris in acid and alkaline soils of Mexico

In order to investigate bean-nodulating rhizobia in different types of soil, 41 nodule isolates from acid and alkaline soils in Mexico were characterized. Based upon the phylogenetic studies of 16S rRNA, atpD, glnII, recA, rpoB, gyrB, nifH and nodC genes, the isolates originating from acid soils were identified as the phaseoli symbiovar of the Rhizobium leguminosarum-like group and Rhizobium grahamii, whereas the isolates from alkaline soils were defined as Ensifer americanum sv. mediterranense and Rhizobium radiobacter. The isolates of “R. leguminosarum” and E. americanum harbored nodC and nifH genes, but the symbiotic genes were not detected in the four isolates of the other two species. It was the first time that “R. leguminosarum” and E. americanum have been reported as bean-nodulating bacteria in Mexico. The high similarity of symbiotic genes in the Rhizobium and Ensifer populations showed that these genes had the same origin and have diversified recently in different rhizobial species. Phenotypic characterization revealed that the “R. leguminosarum” population was more adapted to the acid and low salinity conditions, while the E. americanum population preferred alkaline conditions. The findings of this study have improved the knowledge of the diversity, geographic distribution and evolution of bean-nodulating rhizobia in Mexico.     

Screening seed of Trigonella and three related genera for diosgenin

Seed of 45 species of Leguminosae were analyzed for diosgenin by GLC. Since the method did not separate yamogenin and tigogenin from diosgenin, results indicate maximum limits rather than the actual amounts of diosgenin present. Among 27 species of Trigonella tested, only 6 have as much as 0·27% diosgenin; the highest was 0·8% in 1 sample of T. foenum-graecum. Among 7 species of Medicago, 6 species of Melilotus and 5 species of Trifolium, none contained more than 0·1% diosgenin.     

The taxonomic significance of the fatty acid content of seeds of Linum

About sixty taxa in the genusLinum are arranged systematically and are compared using data published earlier for the fatty acid content of their seeds. Plants of sect.Linum, of sect.Dasylinum, and Old World species of sect.Linastrum generally have higher percentages of linolenic acid and lower percentages of linoleic acid; plants of sect.Syllinum, of sect.Cathartolinum, and New World species of sect.Linastrum have the reverse tendencies. The segregation ofL. stelleroides from other species of sect.Linum and the maintenance ofL. tenuifolium in a section separate from other tested species are supported.