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Molecular diversity of rhizobia nodulating the invasive legume Cytisus scoparius in Australia.

AIMS: To contribute to the understanding of Cytisus scoparius success at invading and establishing itself in Australia. METHODS AND RESULTS: Root-nodule bacteria isolated from C. scoparius, growing on five different sites and originally introduced to Australia, were compared with isolates from indigenous plants growing in France and isolates from native legumes growing on the same Australian sites as C. scoparius. Small-subunit rDNA from 251 isolates were analysed by PCR-RFLP and representatives from different genospecies were selected for sequencing. Phylogenetic analyses revealed a great diversity of lineages belonging to Bradyrhizobium, with one genospecies being specific for Cytisus both in Australia and in France, Rhizobium and Mesorhizobium and one falling outside the described genera of legume-nodulating bacteria. Principal component analysis showed that the Cytisus Australian rhizobial communities are more similar to each other than to their co-occurring native partners. CONCLUSIONS: Early established rhizobial symbionts may have an increased probability to contribute inoculum for the development of further nodules. SIGNIFICANCE AND IMPACT OF THE STUDY: This is a first report comparing rhizobia nodulating C. scoparius in its native and exotic environments. Cytisus scoparius symbionts were identified outside the Bradyrhizobium genus and a new lineage of legume-nodulating bacteria was identified.

Australia↗

Characterization of phages virulent for Sarothamnus scoparius bradyrhizobia.

Four virulent phages--PhiDl, PhiTl, PhiCYT21, and PhiOS6, infective on Sarothamnus scoparius rhizobia--were isolated from the soil and characterized for morphology, host range, rate of adsorption to bacterial cells, and genome size. New phages were separated into two morphological families: Siphoviridae with long, noncontractile tails (PhiDl, PhiTl) and Myoviridae with long, contractile tails (PhiCYT21, PhiOS6). They were also classified into two groups by a host specificity. One of them included viruses (PhiDl and PhiTl) that lysed S. scoparius bradyrhizobia and Bradyrhizobium sp. (Lupinus) strain Dl, and the second one comprised phages (PhiCYT21 and PhiOS6) that parasitized only Scotch broom native microsymbionts. Phages specific for S. scoparius rhizobia were differentiated not only by morphology and host range but also by a genome size that was in the range from 47,583 to 60,098 b.p.

Bradyrhizobiaceae↗

Analysis of genetic relationship of Sarothamnus scoparius microsymbionts and Bradyrhizobium sp. by hybridization in microdilution wells.

DNA-DNA hybridization in microdilution wells was successfully used to determine the overall genomic similarity among Sarothamnus scoparius microsymbionts isolated from Poland and Japan and representatives of Bradyrhizobium species including Bradyrhizobium sp. (Lupinus) USDA3045. Geographically different S. scoparius rhizobia diverged into two genomospecies at the DNA-DNA similarity level of approximately 49%. Polish isolates exhibited high DNA similarity levels to the DNA of Bradyrhizobium japonicum strains (approximately 78%) and formed a common genomospecies. Japanese S. scoparius rhizobia were allocated into a new genomic species due to the low similarity levels of their DNA to the DNA of representative strains of the Bradyrhizobium genus (from 19% to 52%).

Journal Article↗

The root-nodule symbiosis between Sarothamnus scoparius L. and its microsymbionts.

When nitrogen fixing root nodules are formed, Sarothamnus scoparius (broom) is inoculated with its microsymbionts. Nodules studied under light and electron microscopy exhibited typical indeterminate nodule histology with apical, persistent meristem, age gradient of nodule tissues, and open vascular bundles, and also with some particular features such as: the presence of mitotic activity in the infected meristematic cells, lack of infection threads, distribution of bacteria by process of host cell division, and occurrence of a large bacteroid zone only with infected cells. The results of cross-inoculation tests have shown a broad host range for S. scoparius microsymbionts including not only the native host but also species such as: Lupinus luteus, Ornithopus sativa, Lotus corniculatus, Genista tinctoria, Chamaecitisus ratisbonensis, Macroptilium atropurpureum, and Phaseolus vulgaris. In addition, our data established a close symbiotic relationship of S. scoparius nodulators to Bradyrhizobium sp. (Lupinus) by comparison of the partial sequence of nodC gene of the strain CYT7, specific for the broom, to those from Bradyrhizobium sp. (Lupinus) strain D1 and others available in the public databases.

Bacterial Proteins↗

Cytisus scoparius link--a natural antioxidant.

BACKGROUND: Recent investigations have shown that the antioxidant properties of plants could be correlated with oxidative stress defense and different human diseases. In this respect flavonoids and other polyphenolic compounds have gained the greatest attention. The plant Cytisus scoparius contains the main constituent of flavone and flavonals. The present study was undertaken to evaluate the in vitro antioxidant activities of extract of aerial part of Cytisus scoparius. METHODS: The plant extract was tested for DPPH (1, 1-diphenyl, 2-picryl hydrazyl) radical scavenging, nitric oxide radical scavenging, superoxide anion radical scavenging, hydroxyl radical scavenging, antilipid peroxidation assay, reducing power and total phenol content. RESULTS: The extract exhibited scavenging potential with IC50 value of 1.5 microg/ml, 116.0 microg/ml and 4.7 microg/ml for DPPH, nitric oxide and superoxide anion radicals. The values were found to lesser than those of vitamin C, rutin, and curcumin, as standards. The extract showed 50% protection at the dose of 104.0 microg/ml in lipid peroxidation induced by Fe2+/ ascorbate system in rat liver microsomal preparation. There is decrease in hydroxyl radical generation with IC50 value of 27.0 microg/ml when compared with standard vitamin E. The reducing power of the extract depends on the amount of extract. A significant amount of polyphenols could be detected by the equivalent to 0.0589 microg of pyrocatechol from 1 mg of extract. CONCLUSION: The results obtained in the present study indicate that hydro alcoholic extract of aerial part of Cytisus scoparius is a potential source of natural antioxidants.

Animals↗

Nonlocal transplantation and outbreeding depression in the subshrub Lotus scoparius (Fabaceae).

The genetic background of transplants used to create or augment wild populations may affect the long-term success of restored populations. If seed sources are from differently adapted populations, then the relative performance of progeny from crosses among populations may decrease with an increase in genetic differences of parents and in the differences of parental environments to the transplant location. We evaluated the potential for such outbreeding depression by hybridizing individuals from six different populations of Lotus scoparius var. scoparius and L. s. var. brevialatus. We used allozyme data to calculate genetic distances between source populations, and compiled climatic data and measured soil traits to estimate environmental distances between source populations. We found significant outbreeding depression following controlled crosses. In the greenhouse, the success of crosses (seeds/flower × seedlings/seed) decreased with increasing genetic distance between populations revealing genetically based outbreeding depression unrelated to local adaptation. After outplanting to one native site (in situ common garden), field cumulative fitness of progeny (survival × fruit production) decreased significantly with mean environmental distance of the parental populations to the transplant site, but not with genetic distance between the crossed populations. This result is consistent with a disruption of local adaptation. At the second, ecologically contrasting common garden, where low survival reduced statistical power, field cumulative fitness (survival × progeny height) did not decrease significantly with either environmental distance or genetic distance. Overall, intervariety crosses were 40 and 50% as fit (seeds/flower × seedlings/seed × survival × fruits at the first garden or × height at the second) as intravariety crosses. These results suggest that the cumulative outbreeding depression was caused by a combination of genetically based ecological differences among populations and other genomic coadaptation. We conclude that mixing genetically differentiated seed sources of Lotus scoparius may significantly lower the fitness of augmented or restored populations. Genetic and environmental similarities of source populations relative to the transplant site should be considered when choosing source materials, a practice recommended by recent seed transfer policies. Geographic separation was not a good surrogate for either of these measures.

Journal Article↗

Numerical taxonomy of Sarothamnus scoparius rhizobia.

Twenty nodule isolates from Sarothamnus scoparius (broom) growing in Poland and nine strains from plants growing in Japan were studied for phenotypic properties, plasmid presence, phage sensitivity, and host plant specificity. By numerical analysis of phenotypic properties, it was found that the studied nodule bacteria, originating from geographically different countries, constitute two separate groups affiliated to the bradyrhizobium cluster. The membership of S. scoparius rhizobia in the Bradyrhizobium genus was also supported by their long generation time, alkaline reaction in YEM medium with mannitol, lack of plasmids, and wide host plant range.

Bradyrhizobium↗

Antioxidant effect of Cytisus scoparius against carbon tetrachloride treated liver injury in rats.

The study was aimed to investigate the antioxidant activity of Cytisus scoparius L. (Family: Leguminosae) on CCl(4) (carbon tetrachloride) treated oxidative stress in Wistar albino rats. CCl(4) injection induced oxidative stress by a significant rise in serum glutamate oxaloacetate transaminases (SGOT), serum glutamate pyruvate transaminases (SGPT), lactate dehydrogenase (LDH) and thiobarbituric acid reactive substances (TBARS) along with reduction of superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), glutathione peroxidase (GPx), glutathione-s-transferase (GST) and glutathione reductase (GRD). Pretreatment of rats with different doses of plant extract (250 and 500mg/kg) significantly lowered SGOT, SGPT, LDH and TBARS levels against CCl(4) treated rats. GSH and hepatic enzymes like SOD, CAT, GPx, GRD, and GST were significantly increased by treatment with the plant extract, against CCl(4) treated rats. The activity of extract at the dose of 500mg/kg was comparable to the standard drug, silymarin (25mg/kg). Based on these results, it was observed that Cytisus scoparius extract protects liver from oxidative stress induced by CCl(4) in rats and thus helps in evaluation of the traditional claim on this plant.

Alanine Transaminase↗

Possible source of tetrodotoxin in the starfish Astropecten scoparius.

The seasonal variations of toxicity and stomach contents in toxic starfish Astropecten scoparius were detected. The average highest specimen toxicity, expressed as tetrodotoxin (TTX), was 16,821 mouse units (MU). The toxin was composed of TTX only, except April's sample containing mainly TTX along with minor paralytic shellfish poisons. The composition in the stomach of less and more toxic starfish was mainly Veremolpa scabra and Umborium suturale, respectively. The toxicities of U. suturale and V. scabra, found in the starfish stomach were 65 and 33 MU, respectively. For those collected from the coastal waters, U. suturale was toxic with average toxicity value of 77 MU/g, but V. scabra was nontoxic. The toxin in the specimens of U. suturale collected from either the digestive gland of starfish or the coastal waters, was TTX and anhydroTTX only. It indicates that the starfish A. scoparius might mainly accumulate high amount of TTX from U. suturale. Furthermore, both small gastropods U. suturale and Natica psuestes are first reported to contain TTX.

Animals↗

The fucosyl specific lectins of Ulex europaeus and Sarothamnus scoparius. Biochemical characteristics and binding properties to human B-lymphocytes.

From the seeds of the gorse, Ulex europaeus and of the broom, Sarothamnus scoparius L-fucosyl-specific lectins were isolated by affinity chromatography on L-fucosyl-epoxy-Sepharose. The lectins showed similarities in their molecular weights, amino acid composition, carbohydrate content and in the finger-prints of their tryptic peptides. The fluorescein-labeled lectins of both seeds attached especially to the plasma membranes of human B-lymphocytes.

Amino Acids↗

Paralytic toxins in Taiwanese starfish Astropecten scoparius.

Paralytic toxicity was seasonally detected in every 10 specimens of the starfish Astropecten scoparius from Pingtung in Taiwan from December 1995 to November 1996. The highest toxicity and the average toxicity of total specimens, expressed as tetrodotoxin (TTX), were 5,938 and 1,679 mouse unit (MU), respectively. The highest value of monthly average toxicity in viscera and other parts were 354 and 247 MU/g, respectively. The toxin was partially purified by YM-2 membrane ultrafiltration and Bio-Gel P-2 column chromatography. HPLC and GC-MS analyses showed that the starfish toxin was mainly composed of tetrodotoxin (88%), along with minor gonyautoxin 2-3 and saxitoxin (12%).

Animals↗

Structural differences between two lectins from Cytisus scoparius, both specific for D-galactose and N-acetyl-D-galactosamine.

Three lectin fractions were obtained from seeds of the leguminous plant Cytisus scoparius (Scotch broom) by means of affinity chromatography on a N-acetyl-D-galactosamine medium. The first fraction, termed CSIa, was equally well inhibited in haemagglutination experiments by D-galactose and by N-acetyl-D-galactosamine and consisted of a group of isolectins formed from closely related polypeptide chains of approx. Mr 30000. The second fraction, CSIb, was closely related to CSIa in specificity, c.d. and other properties. The third fraction contained a homogeneous lectin, CSII, formed from subunits again of approx. Mr 30000. CSII was 100 times more readily inhibited by N-acetyl-D-galactosamine than by D-galactose. Despite the similarity in specificity, comparative studies of their amino acid composition, c.d. and N-terminal amino acid sequence showed that the CSIa and CSII lectins diverged considerably in structure. The lectin from Cytisus sessilifolius, specific for chitobiose, was also examined and resembled CSIa in composition and c.d. properties.

Acetylgalactosamine↗

Alkylated isocoumarins from Pituranthos scoparius.

Two isocoumarins have been isolated from the ethyl acetate extract of the root of Pituranthos scoparius: 3-n-propyl-5-methoxy-6-hydroxy-isocoumarin and 3-n-propyl-5,7-dimethoxy-6-hydroxy-isocoumarin. Their structures were assigned by spectral analysis.

Apiaceae↗

The primary structure of the Cytisus scoparius seed lectin and a carbohydrate-binding peptide.

The complete amino acid sequence of 2-acetamido-2-deoxy-D-galactose-binding Cytisus scoparius seed lectin II (CSII) was determined using a protein sequencer. After digestion of CSII with endoproteinase Lys-C or Asp-N, the resulting peptides were purified by reversed-phase high performance liquid chromatography (HPLC) and then subjected to sequence analysis. Comparison of the complete amino acid sequence of CSII with the sequences of other leguminous seed lectins revealed regions of extensive homology. The amino acid residues of concanavalin A (Con A) involved in the metal binding site are highly conserved among those of CSII. A carbohydrate-binding peptide of CSII was obtained from the endoproteinase Asp-N digest of CSII by affinity chromatography on a column of GalNAc-Gel. This peptide was retained on the GalNAc-Gel column and was presumed to have affinity for the column. The amino acid sequence of the retarded peptide was determined using a protein sequencer. The retarded peptide was found to correspond to the putative metal-binding region of Con A. These results strongly suggest that this peptide represents the carbohydrate-binding and metal ion-binding sites of CSII.

Amino Acid Sequence↗

Development and metamorphosis of the sea-star, Astropecten scoparius Valenciennes.

1. Astropecten scoparius develops to a bipinnaria with simple ciliary bands and short bipinnari arms through a wrinkled blastula by holoblastic, radial cleavage. 2. About seven days after insemination, the posterior portion of the bipinnaria becomes swollen and fine spicules appear on it, while the anterior portion, the stalk, remains unchanged. 3. Metamorphosis takes place gradually at the posterior portion, while the metamorphosing bipinnaria is pelagic. Two weeks after insemination, the stalk rapidly shrinks and the larva sinks to the bottom. 4. About 18 days after insemination, the juvenile completes metamorphois with the opening of the mouth. The newly metamorphosed juvenile is 600 mu in diameter and each arm bears two pairs of the tube-feet, each having a sucker at the tip, and one terminal tentacle with red eye-spot. 5. The aboral skeletal system of the juvenile immediately after metamorphosis is composed of one central, one madreporic, ten radial and interradial plates, in addition to five terminal plates on the arms. 6. The juveniles smaller than about one cm in R do not bear some of the diagnostic features in this species. 7. A characteristic feature of the development of Astropecten, i.e., the lack of a brachiolaria stage, is stressed. The term "nonbrachiolarian type" is tentatively proposed to distinguish the development of Astropecten and Luidia, which do not pass through a brachiolaria stage, from the usual indirect type of development.

Animals↗

Role of specialized microvilli and the fertilization envelope in the spatial positioning of blastomeres in early development of embryos of the starfish Astropecten scoparius.

In the eggs of a wide range of animal species, various factors that determine the blastomeres' presumptive fate are known to locate unevenly within the egg. In the embryos of these animals, cleavage occurs not just to increase cell numbers, but also to distribute the factors to the respective blastomeres, resulting in cell specialization at the later stages. In the early cleavage stages, before the establishment of a device such as desmosomes to directly join the blastomeres, some other means is needed to keep the blastomeres together and maintain the relative positions among them. In this study, we found that the embryos of the starfish Astropecten scoparius lack the hyaline layer seen in sea urchin embryos and that blastomeres adhere to the fertilization envelope (FE) via filamentous cellular projections (fixing processes). Electron microscopy revealed the fixing processes to be specialized microvilli formed, after the elevation of the FE, by the elongation of short microvilli that pre-exist in unfertilized eggs. After the first cleavage, the two blastomeres separate from each other and finally attach to the FE. In the subsequent cleavages, the blastomeres undergo repeated cell division without separating from the FE. Between the blastomeres and the FE, only shortened fixing processes were observed. Destruction of the fixing processes caused release of the blastomeres from the FE and disturbance of the relative positions of the blastomeres, resulting in abnormal development of the embryos. These observations suggest that the fixing process is a device to keep the egg placed centrally in the FE up to the first cleavage, and after the first cleavage and beyond to anchor the blastomeres to the FE so that the FE can be used as a scaffold for morphogenesis. Electron microscopy also suggests that the inner layer of the FE, which is derived from the contents of cortical granules, reinforces the adhesion of the fixing processes to the FE. Immuno-electron microscopy, using an antibody against sea urchin hyaline layer, showed that the inner layer of the FE of starfish eggs and the hyaline layer of sea urchin eggs, which are both derived from cortical granules, contain some common elements.

Animals↗