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Diversity of frankia strains in root nodules of plants from the families elaeagnaceae and rhamnaceae

Partial 16S ribosomal DNAs (rDNAs) were PCR amplified and sequenced from Frankia strains living in root nodules of plants belonging to the families Elaeagnaceae and Rhamnaceae, including Colletia hystrix, Elaeagnus angustifolia, an unidentified Elaeagnus sp., Talguenea quinquenervia, and Trevoa trinervis. Nearly full-length 16S rDNAs were sequenced from strains of Frankia living in nodules of Ceanothus americanus, C. hystrix, Coriaria arborea, and Trevoa trinervis. Partial sequences also were obtained from Frankia strains isolated and cultured from the nodules of C. hystrix, Discaria serratifolia, D. trinervis, Retanilla ephedra, T. quinquenervia, and T. trinervis (Rhamnaceae). Comparison of these sequences and other published sequences of Frankia 16S rDNA reveals that the microsymbionts and isolated strains from the two plant families form a distinct phylogenetic clade, except for those from C. americanus. All sequences in the clade have a common 2-base deletion compared with other Frankia strains. Sequences from C. americanus nodules lack the deletion and cluster with Frankia strains infecting plants of the family Rosaceae. Published plant phylogenies (based on chloroplast rbcL sequences) group the members of the families Elaeagnaceae and Rhamnaceae together in the same clade. Thus, with the exception of C. americanus, actinorhizal plants of these families and their Frankia microsymbionts share a common symbiotic origin.

Journal Article↗

Wood anatomy of Elaeagnaceae, with comments on vestured pits, helical thickenings, and systematic relationships.

The secondary xylem of Elaeagnus, Hippophae, and Shepherdia is described and illustrated in detail. Shrubs and small trees of Elaeagnaceae have ring-porous or semi-ring-porous wood with simple perforation plates, vascular tracheids, fiber-tracheids, diffuse or rarely paratracheal axial parenchyma, and uni- or biseriate rays in Hippophae and Shepherdia, but wider rays in Elaeagnus. Walls of vessel elements, especially narrow ones, tracheids, or fiber-tracheids sometimes show helical thickenings; in a few instances these intergrade with small bud-like protrusions associated with pits. Scanning electron microscopy illustrates that small to vestigial vestures are present in all species studied, although nonvestured pits are also common. The analogous nature of vestures and helical thickenings is considered. Comparative wood anatomy suggests a rather isolated position of the family Elaeagnaceae; affinities with Rhamnaceae, Proteaceae, and Thymelaeaceae are discussed.

Journal Article↗

Genetic variation in Hippophae rhamnoides ssp. sinensis (Elaeagnaceae) revealed by RAPD markers.

Hippophae rhamnoides ssp. sinensis is endemic to China, and it is a dioecious, outcrossing plant. Although many studies have been undertaken mainly on its agricultural, nutritional, medical, and ornamental value, little is known about its population genetics. This study uses random amplified polymorphic DNA to investigate the genetic diversity and population genetic structure of 13 natural populations of the subspecies sinensis. Fifteen primers amplified 107 reproducible bands, with 95 (88.79%) being polymorphic. The gene diversity within population was 0.168, considerably lower than that of tree species and most perennial, outcrossing species, but higher than that of annual or short-lived, selfing species. The Gst value showed that 18.3% of the total genetic variation resided among populations, a little lower than that of outcrossing species. The present results are quite similar to those previously reported in another subspecies, H ssp. . rhamnoides rhamnoides. The low genetic differentiation among populations in ssp. sinensis may be attributed to the long-distance dispersal of seeds facilitated by birds, in addition to its characteristics of outcrossing, wind pollination, and widespread distribution. No association between genetic distance and geographical distribution was found. The population relationships revealed by the UPGMA dendrogram parallel this result, in that genetic distance did not increase with geographic separation. This pattern of population differentiation may imply the adaptation of ssp. s populations to the local environment, given that its habitats vary greatly across its distribution.

China↗

Late Quaternary history of Hippophaë rhamnoides L. (Elaeagnaceae) inferred from chalcone synthase intron (Chsi) sequences and chloroplast DNA variation.

Fossil pollen records indicate that Hippophaë rhamnoides (Sea Buckthorn) was widespread on late- and early postglacial raw soils throughout much of central and northern Europe, but that Early Holocene reforestation restricted populations to northern coastal habitats, or along mountain streams in the Alps, Pyrenees, and Carpathians. We used sequence variation at the nuclear chalcone synthase intron (Chsi), in conjunction with chloroplast DNA-restriction fragment length polymorphism data, to investigate the intraspecific phylogeny, phylogeographic structure, and expansion demographic history of this dioecious and wind-pollinated shrub at its range-wide scale in Europe and Asia Minor. Four major Chsi phylogroups of unresolved relationships were identified with estimated divergences approximately 172,000 years ago. Large-scale phylogeographic structures of nuclear and cytoplasmic markers were congruent in identifying (i) southeastern Europe as the most likely source of colonization into central Europe and Scandinavia, and (ii) the area just north of the Alps as a contact zone between populations from the Alps and the east/central European-Scandinavian lineage. Coalescence-based analyses (i.e. nested clade analysis and mismatch distributions) of Chsi variation were able to detect at least four major episodes of population growth, all within about the last 40,000 years. In particular, these analyses identified a nearly synchronized timing of population expansions in various parts of the species' range in central-eastern Europe/Asia Minor, most likely correlating with the Younger Dryas Stadial ( approximately 13,000-11,600 years ago). It remains to be established whether the phylogeographic history of H. rhamnoides, and particularly its rapid response to the rapid environmental changes of the Younger Dryas cold snap, is unique to the species, or whether it is shared with other cold-tolerant shrub (or grassland) species known from late-glacial raw soils in Europe.

Acyltransferases↗

Isolation of Elaeagnus-compatible Frankia from soils collected in Tunisia.

The occurrence and diversity of Frankia nodulating Elaeagnus angustifolia in Tunisia were evaluated in 30 soils from different regions by a Frankia-capturing assay. Despite the absence of actinorhizal plants in 24 of the 30 soils, nodules were captured from all the samples. Eight pure strains were isolated from single colonies grown in agar medium. On the basis of 16S rRNA and GlnII sequences, seven strains were clustered with Frankia, colonizing Elaeagnaceae and Rhamnaceae in two different phylogenetic groups while one strain described a new lineage in the Frankia assemblage, indicating that Frankia strains genetically diverse from previously known Elaeagnus-infective strains are present in tunisian soils. Genomic fingerprinting determined by rep-PCR, and tDNA-PCR-SSCP, confirmed the wide genetic diversity of the strains.

Base Sequence↗

Muscle relaxant activity of Elaeagnus angustifolia L. fruit seeds in mice.

Muscle relaxant effect of Elaeagnus angustifolia L. (Elaeagnaceae) fruit seeds was studied in mice using traction test. The aqueous and ethanolic extracts (i.p) induced a muscle relaxant effect in a dose dependent manner as effective as diazepam (1 mg/kg). The aqueous extract was partitioned with methanol-chloroform (MeCh) and n-butanol (Bu.) saturated with water. The MeCh and Bu. fractions did not show activity. Preliminary phytochemical tests showed that the extract contains flavonoid. The results suggested that E. angustifolia fruit seeds exerted muscle relaxant activity via flavonoid component(s).

Animals↗

Anti-ulcerogenic activity of some plants used as folk remedy in Turkey.

Five herbal remedies used as gastroprotective crude drugs in Turkey were assessed for anti-ulcerogenic activity using the EtOH-induced ulcerogenesis model in rat. The crude drugs investigated comprises fruits of Elaeagnus angustifolia L. (Elaeagnaceae), fresh fruits of Hibiscus esculentus L. (Malvaceae), fresh roots of Papaver rhoeas L. (Papaveraceae), leaves of Phlomis grandiflora H.S. Thomson (Lamiaceae) and fresh fruits of Rosa canina L. (Rosaceae). Extracts were prepared according to the traditional indications of use. Under our experimental conditions, all extracts exhibited statistically significant gastroprotective effect with better results for Phlomis grandiflora and Rosa canina (100%). At the concentration under study, both crude drugs were more effective than the reference compound misoprostol at 0.4 mg/kg. Even the weakest anti-ulcerogenic effect observed for Papaver rhoeas roots was found statistically potent (95.6%). Histopathological studies confirmed the results of the in vivo test.

Animals↗

Distribution and contents of phenolic compounds in eighteen Scandinavian berry species.

Berries contain a wide range of phenolic compounds in different conjugated forms, a fact that makes their simultaneous analysis a difficult task. In this work, soluble and insoluble phenolic compounds were identified and quantified in 18 species of berries by reversed phase high-performance liquid chromatography combined with diode array detection. The analytical results and literature data were used for the identification of the predominant conjugated hydroxycinnamic acids, flavonol glycosides, and anthocyanins in berries from six families, viz. Grossulariaceae, Ericaceae, Rosaceae, Empetraceae, Elaeagnaceae, and Caprifoliaceae. The study showed distinctive similarities among berry species of the same family in the distribution of conjugated forms of phenolic compounds but differences in chromatographic profiles of conjugates and compositions of aglycones especially in the case of anthocyanins. The chromatographic profiles of chokeberry and the related sweet rowanberry (Rosaceae) were exceptionally similar. These data are informative to studies on the authenticity of berry raw materials as well as to those on the evaluation of berries as sources of phenolic compounds.

Anthocyanins↗

Chemistry and function of vegetable polyphenols with high molecular weights.

Structure and function of polypehnols with high molecular weights (tannins) were briefly reviewed to better understand the significance of polyphenol-rich foods and beverages. In a survey of bioactive ellagiannins with a macrocyclic structure and/or a gluconic acid core, some new oligomeric ellagitannins (eucarpanins and elaeagnatins) have been found in species of Myrtaceae and Elaeagnaceae, and their structures were elucidated by spectroscopic and chemical methods. Cytotoxic activity against human oral tumor cell lines and antibacterial activity against Helicobacter pylori have been evaluated for the ellagitannins obtained from both plants, and related compounds. The macrocyclic dimers, oentothein B, camelliin B and woodfordin C showed a remarkable cytotoxicity against human oral squamous cell carcinoma, but not against normal cells. These active tannins induced apoptosis of tumor cells. A potent antibacterial activity against Helicobacter pylori was exhibited by monomeric ellagitannins such as tellimagrandin I and stricitinin.

Anti-Bacterial Agents↗

Polar lipid and fatty acid distribution in carotenolipoprotein complexes extracted from sea buckthorn fruits.

Sea buckthorn (Hippophae rhamnoides L., fam. Elaeagnaceae) fruits are rich in pigments and lipoproteins located in membranes and the fleshy mesocarp. In spite of many reports concerning the neutral lipids in the mesocarp, no data about the polar lipids and their fatty acid composition are available even though they play important structural and physiological roles in cell membranes and may offer interesting applications as emulsifiers and nutrients in cosmetic preparations. Carotenolipoprotein complexes are located particularly in fruit membranes where polar lipids may function as bridge compounds between the polar (protein) and non-polar (carotenoid) moieties. The fatty acid compositions of total and individual polar lipids separated from carotenolipoprotein complexes were determined by HPTLC and GC. The polar lipids included 61% phospholipids and 39% galactolipids, which contained mainly 16:0, 16:1 (9c), 18:1 (9c), 18:1 (11c) and 18:2 (9c, 12c) fatty acids. Almost all polar lipids showed high ratios of 16:0/16:1 (11c) and 18:1 (9c)/18:1 (11c), and higher quantities of 18 carbon unsaturated fatty acids than of the saturated analogue. Galactolipids proved to be richest in 18:1 (9c) and 18:3 (9c, 12c, 15c) fatty acids, while phospholipids contained higher concentrations of 16:0 and 18:1 (9c).

Carotenoids↗

Phylogenetic relationships of the enigmatic angiosperm family Podostemaceae inferred from 18S rDNA and rbcL sequence data.

The phylogenetic relationships of some angiosperm families have remained enigmatic despite broad phylogenetic analyses of rbcL sequences. One example is the aquatic family Podostemaceae, the relationships of which have long been controversial because of major morphological modifications associated with their aquatic habit. Podostemaceae have variously been associated with Piperaceae, Nepenthaceae, Polygonaceae, Caryophyllaceae, Scrophulariaceae, Rosaceae, Crassulaceae, and Saxifragaceae. Two recent analyses of rbcL sequences suggest a possible sister-group relationship of Podostemaceae to Crassulaceae (Saxifragales). However, the branch leading to Podostemaceae was long, and use of different outgroups resulted in alternative placements. We explored the phylogenetic relationships of Podostemaceae using 18S rDNA sequences and a combined rbcL + 18S rDNA matrix representing over 250 angiosperms. In analyses based on 18S rDNA data, Podostemaceae are not characterized by a long branch; the family consistently appears as part of a Malpighiales clade that also includes Malpighiaceae, Turneraceae, Passifloraceae, Salicaceae, Euphorbiaceae, Violaceae, Linaceae, Chrysobalanaceae, Trigoniaceae, Humiriaceae, and Ochnaceae. Phylogenetic analyses based on a combined 18S rDNA + rbcL data set (223 ingroup taxa) with basal angiosperms as the outgroup also suggest that Podostemaceae are part of a Malpighiales clade. These searches swapped to completion, and the shortest trees showed enhanced resolution and increased internal support compared to those based on 18S rDNA or rbcL alone. However, when Gnetales are used as the outgroup, Podostemaceae appear with members of the nitrogen fixing clade (e.g., Elaeagnaceae, Ulmaceae, Rhamnaceae, Cannabaceae, Moraceae, and Urticaceae). None of the relationships suggested here for Podostemaceae receives strong bootstrap support. Our analyses indicate that Podostemaceae are not closely allied with Crassulaceae or with other members of the Saxifragales clade; their closest relatives, although still uncertain, appear to lie elsewhere in the rosids.

DNA, Ribosomal↗

Genetic diversity of Frankia microsymbionts from the relict species Myrica faya (Ait.) and Myrica rivas-martinezii (S.) in Canary Islands and Hawaii.

In the Western Canary Islands, Myrica faya and Myrica rivas-martinezii (Myricaceae) are phylogenetically close, endemic, actinorhizal species presumed to be remnants either of the European or the African Tertiary floras. Unisolated Frankia strains from field-collected nodules on Tenerife, Gomera, and La Palma Islands were compared by their rrs gene and 16S-23S intergenic spacer (IGS) restriction patterns. To compare the genetic diversity of Frankia strains from within and outside the host's native range, nodules of M. faya field plants were collected both in Canary Islands and in Hawaii, where this species is an exotic invasive. Myrica rivas-martinezii, endemic to the Canary Islands, was sparsely nodulated in the field. Frankia strains harbored in field-collected nodules of M. faya and M. rivas-martinezii belonged to the Elaeagnaceae strains' genetic cluster and exhibited a high degree of diversity. Frankia genotypes were specific to each host species. In the Canary archipelago, we found no relationship between site of collection and Frankia genotype for M. faya. The only exceptions were strains from site 2 in Tenerife, a location with a geological history different from the other sites sampled. Hawaiian and Canarian M. faya strains had no genotypes in common, raising questions concerning the origin of M. faya-infective Frankia in Hawaii. Nodular strains of M. rivas-martinezii from nursery plants were genetically characterized and shown to be divergent from the strains of field-collected nodules and belong to the Alnus-Casuarina strains cluster. This suggests Myrica may have the potential to nodulate with a broader range of Frankia genotypes under artificial conditions than has been detected in field-collected nodules.

Atlantic Islands↗

Arum- and Paris-type arbuscular mycorrhizas in a mixed pine forest on sand dune soil in Niigata Prefecture, central Honshu, Japan.

Arbuscular mycorrhizas (AM) are the most widespread mycorrhiza in nature and form two morphologies, Arum- and Paris-type. The determining factors defining the two different morphologies are not well understood. In this study, the distribution of Arum- and Paris-type AM was determined in a mixed pine forest. A total of 35 plant species belonging to 20 families and 32 genera were identified and examined for AM colonization and morphological types. AM morphological types in 14 families were confirmed as follows: Arum-type in Rosaceae, Oleaceae, Lauraceae, Vitaceae and Compositae, Paris-type in Aquifoliaceae, Ulmaceae, Araliaceae, Theaceae, Magnoliaceae, Rubiaceae and Dioscoraceae, and both and/or intermediate types in Caprifoliaceae and Gramineae. Plant families whose AM morphological status was previously unknown were clarified as follows: Polygonaceae and Commelinaceae showed Arum-type morphology; Celastraceae, Menispermaceae and Elaeagnaceae had typical Paris-type morphology. The proportion of Arum-type to Paris-type species decreased in the following order: annuals > perennials > deciduous species > evergreen species, and pioneer group > early successional group > late successional group. Evergreen plants had a higher tendency to form Paris-type AM than annuals, perennials and deciduous plants. The results indicate that environmental changes, such as shade during plant succession, control the distribution of plant growth forms in mixed pine forest and may also play a part in the distribution of Arum- and Paris-type morphology. The identity of the plant seems to strongly influence AM morphology, though control by the fungal genome cannot be ruled out.

Japan↗

Adaptogenic and safety evaluation of seabuckthorn (Hippophae rhamnoides) leaf extract: a dose dependent study.

The effects of seabuckthorn (Hippophae rhamnoides L., Elaeagnaceae), leaf aqueous extract were examined in rats for its adaptogenic activity and toxicity. Dose dependent adaptogenic study of extract was carried out at different doses administered orally, 30min prior to cold (5 degrees C)-hypoxia (428mmHg)-restraint (C-H-R) exposure. After sub-acute toxicity studies on 10 and 20 times doses of maximal effective dose administered for 14 days (single oral dose of 1g/kg and 2g/kg once daily) and maximal effective dose administered for 30 days (single oral dose of 100mg/kg once daily), biochemical and hematological parameters were studied in the serum and blood. The maximal effective adaptogenic dose of the extract was 100mg/kg body weight. No significant changes were observed in organ weight/body weight ratios, of any vital organ studied (except liver and kidney in 1g/kg and 2g/kg body weight doses, respectively), and biochemical and hematological parameters of the sub-acute drug treated animals in comparison to control rats. In acute toxicity study LD(50) of the extract was observed to be >10g/kg when given orally. These results indicate that seabuckthorn leaf aqueous extract possess potent adaptogenic activity with no toxicity even after sub-acute (30 days) maximal effective dose administration.

Animals↗

In vitro anti-Helicobacter pylori action of 30 Chinese herbal medicines used to treat ulcer diseases.

Infection by Helicobacter pylori has been ascertained to be an important etiologic impetus leading usually to chronic active gastritis and gastric ulcer with growing incidences worldwide. Utilizing as the test pathogen a standard and five clinic strains of Helicobacter pylori, the antibacterial action was assessed in vitro with ethanol extracts of 30 Chinese herbal medicines which have been frequently prescribed since ancient times for treating gastritis-like disorders. Among the 30 tested materials, the ethanol extracts of Abrus cantoniensis (Fabaceae), Saussurea lappa (Asteraceae) and Eugenia caryophyllata (Myrtaceae) were strongly inhibitory to all test strains (MICs: approximately 40 microg/ml), and Hippophae rhamnoides (Elaeagnaceae), Fritillaria thunbergii (Liliaceae), Magnolia officinalis and Schisandra chinensis (Magnoliaceae), Corydalis yanhusuo (Papaveraceae), Citrus reticulata (Rutaceae), Bupleurum chinense and Ligusticum chuanxiong (Apiaceae) substantially active with MICs close to 60.0 microg/ml. As to antibacterial actions of the aqueous extracts of the same drugs, those derived from Cassia obtusifolia (Fabaceae), Fritillaria thunbergii and Eugenia caryophyllata were remarkably inhibitory against all the six Helicobacter pylori strains (MICs: approximately 60 microg/ml). The work compared almost quantitatively the magnitude of the anti-Helicobacter pylori actions of the 30 most prescribed gastritis-treating Chinese herbal drugs, and located as well some source plants where potent anti-Helicobacter pylori phytochemicals could be characterized.

Anti-Bacterial Agents↗

Molecular phylogeny of Myricaceae: a reexamination of host-symbiont specificity.

The phylogeny of 13 species of Myricaceae, the most ancient actinorhizal family involved in a nitrogen-fixing symbiosis with the actinomycete Frankia, was established by the analysis of their rbcL gene and 18S-26S ITS. The phylogenetic position of those species was then compared to their specificity of association with Frankia in their natural habitat and to their nodulation potential determined on greenhouse-grown seedlings. The results showed that Genus Myrica, including M. gale and M. hartwegii, and Genus Comptonia, including C. peregrina, belong to a phylogenetic cluster distinct from the other Myrica species transferred in a new genus, Morella. This grouping parallels the natural specificity of each cluster with Comptonia-Myrica and Morella being nodulated by two phylogenetically divergent clusters of Frankia strains, the Alnus and Elaeagnaceae-infective strains clusters, respectively. Under laboratory conditions, Comptonia and Morella had a nodulation potential larger than under natural conditions. From this study it appears that the Myricaceae are split into two different specificity groups. It can be hypothesized that the early divergence of the genera led to the selection of genetically diverse Frankia strains which is contradictory to the earlier proposal that evolution has proceeded toward narrower promiscuity within the family.

Evolution, Molecular↗