Efficient separation and purification of allophycocyanin from <Emphasis Type=Italic>Spirulina</Emphasis> (<Emphasis Type=Italic>Arthrospira</Emphasis>) <Emphasis Type=Italic>platensis</Emphasis>

Allophycocyanin (APC) is a minor component of phycobiliproteins in cyanobacteria and red algae. This paper describes a simple and inexpensive extracting method for isolating APC from Spirulina (Arthrospira) platensis with high efficiency. The crude phycobiliprotein extract was pretreated by ammonium sulfate fractionation. Then, by adding hydroxylapatite into crude phycobiliprotein extract dissolved in 20 mM phosphate buffer (pH 7.0), APC was selectively adsorbed by hydroxylapatite but C-phycocyanin (C-PC) was not. The hydroxylapatite was collected and APC was extracted from the crude phycobiliprotein extract. Then, the enriched APC was washed off from the hydroxylapatite using 100 mM phosphate buffer (pH 7.0). In this simple extracting method it was easy to remove C-PC and isolate APC in large amounts. The absorbance ratio A ₆₅₀/A ₂₈₀ of extracted APC reached 2.0. The recovery yield was 70%, representing 4.61 mg · g⁻¹ wet weight. The extracted APC could be further purified by a simple anion-exchange chromatography with a pH gradient from 5.6 to 4.0. The absorbance ratio A ₆₅₀/A ₂₈₀ of the purified APC reached 5.0, and the overall recovery yield was 43%, representing 2.83 mg · g⁻¹ wet weight. Its purity was confirmed by native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate-PAGE.     

Cryopreservation of <Emphasis Type=Italic>Robinia</Emphasis><Emphasis Type=Italic>pseudoacacia</Emphasis>

Cryopreservation of Robinia pseudoacacia explants by vitrification achieved 78% survival following the stepwise preculture of shoot tips in (0.3 + 0.5 + 0.7 M) sucrose with a 80 min incubation in PVS2; compared to 87% survival after desiccation of explants to 30% water content, following 3 days alginate bead (with glycerol and sucrose treatments) preculture in 0.7 M sucrose.     

New species of <Emphasis Type=Italic>Dypsis</Emphasis> and <Emphasis Type=Italic>Ravenea</Emphasis> (Arecaceae) from Madagascar

Fourteen new species of palms (Arecaceae) from Madagascar are described and named, based on material collected over the last 15 years. Twelve species belong to the genus Dypsis, namely D. andilamenensis Rakotoarin. & J. Dransf., D. anjae Rakotoarin. & J. Dransf., D. betsimisarakae Rakotoarin. & J. Dransf., D. culminis Rakotoarin. & J. Dransf., D. dracaenoides Rakotoarin. & J. Dransf., D. gautieri Rakotoarin. & J. Dransf., D. gronophyllum Rakotoarin. & J. Dransf., D. jeremiei Rakotoarin. & J. Dransf., D. metallica Rakotoarin. & J. Dransf., D. reflexa Rakotoarin. & J. Dransf., D. sancta Rakotoarin. & J. Dransf. and D. vonitrandambo Rakotoarin. & J. Dransf. and two species belong to the genus Ravenea: R. beentjei Rakotoarin. & J. Dransf. and R. hypoleuca Rakotoarin. & J. Dransf. Despite the fact that most of these species have been recorded from protected areas that are difficult to access in the eastern region of Madagasacar, they are all threatened. Based on IUCN categories and criteria, seven are Critically Endangered and seven are Vulnerable.     

<Emphasis Type=Italic>Agrobacterium tumefaciens</Emphasis>-mediated transformation of oleaginous yeast <Emphasis Type=Italic>Lipomyces</Emphasis> species

Interest in using renewable sources of carbon, especially lignocellulosic biomass, for the production of hydrocarbon fuels and chemicals has fueled interest in exploring various organisms capable of producing hydrocarbon biofuels and chemicals or their precursors. The oleaginous (oil-producing) yeast Lipomyces starkeyi is the subject of active research regarding the production of triacylglycerides as hydrocarbon fuel precursors using a variety of carbohydrate and nutrient sources. The genome of L. starkeyi has been published, which opens the door to production strain improvements through the development and use of the tools of synthetic biology for this oleaginous species. The first step in establishment of synthetic biology tools for an organism is the development of effective and reliable transformation methods with suitable selectable marker genes and demonstration of the utility of the genetic elements needed for expression of introduced genes or deletion of endogenous genes. Chemical-based methods of transformation have been published but suffer from low efficiency. To address these problems, Agrobacterium-mediated transformation was investigated as an alternative method for L. starkeyi and other Lipomyces species. In this study, Agrobacterium-mediated transformation was demonstrated to be effective in the transformation of both L. starkeyi and other Lipomyces species. The deletion of the peroxisomal biogenesis factor 10 gene was also demonstrated in L. starkeyi. In addition to the bacterial antibiotic selection marker gene hygromycin B phosphotransferase, the bacterial β-glucuronidase reporter gene under the control of L. starkeyi translation elongation factor 1α promoter was also stably expressed in six different Lipomyces species. The results from this study demonstrate that Agrobacterium-mediated transformation is a reliable and effective genetic tool for homologous recombination and expression of heterologous genes in L. starkeyi and other Lipomyces species.     

<Emphasis Type=Italic>Barleria compacta</Emphasis>: a new species in <Emphasis Type=Italic>Barleria</Emphasis> sect. <Emphasis Type=Italic>Prionitis</Emphasis> (Acanthaceae) from Somalia

Following a re-examination of the material treated under Barleria brevispina (Fiori) Hedrén in the recent Flora of Somalia account of the Acanthaceae, it is concluded that two distinct species are involved and Barleria compacta Malombe & I. Darbysh. is described here from north-eastern Somalia. Its affinities and conservation status are discussed.     

Attenuation of fibrosis <Emphasis Type=Italic>in vitro</Emphasis> and <Emphasis Type=Italic>in vivo</Emphasis> with <Emphasis Type=Italic>SPARC</Emphasis> siRNA

Introduction  

SPARC is a matricellular protein, which, along with other extracellular matrix components including collagens, is commonly over-expressed in fibrotic diseases. The purpose of this study was to examine whether inhibition of SPARC can regulate collagen expression in vitro and in vivo, and subsequently attenuate fibrotic stimulation by bleomycin in mouse skin and lungs.     

<Emphasis Type=Italic>Boletus kermesinus</Emphasis>, a new species of <Emphasis Type=Italic>Boletus</Emphasis> section <Emphasis Type=Italic>Luridi</Emphasis> from central Honshu,Japan

Boletus kermesinus, a new species of Boletus section Luridi, is fully described and illustrated based on the materials collected in subalpine coniferous forests of central Honshu, Japan. It has distinctive features of dark-red basidiomata having distinct viscidity in the pileus surface, usually unchanging flesh, discolorous red pores, and an entirely reticulate stipe becoming coarsely lacerate-rimose with age.     

<Emphasis Type=Italic>In vitro</Emphasis> propagation of ten threatened species of <Emphasis Type=Italic>Mammillaria</Emphasis> (Cactaceae)

Mammillaria species are the most numerous within Cactaceae family, and some of them are threatened with extinction as a result of human activities. In this work, results of in vitro propagation are presented for ten Mammillaria species, testing 20 combinations of indole-3-acetic acid (IAA) and kinetin. Best results on shoot formation were obtained using kinetin at two levels: 27.9 and 46.5 μM. All IAA levels tested were able to induce de novo shoot formation in M. bocasana, M. densispina, M. hahniana, M. hutchisoniana, M. orcutii, M. pectinifera, M. perbella, M. picta, M. rhodantha, and M. zephyranthoides. Depending on the IAA level tested, four responding groups were observed concerning their highest shoot-formation number. For all species, the highest average of shoot formation was achieved with 5.7:46.5 or 11.4:46.5 μM IAA/kinetin, yielding 4.8 and 4.7 shoots per explant, respectively, in 60 d. Rooting of regenerated shoots was achieved by leaving the explants in their shoot-induction medium or transferring them to half-strength MS medium. Hardening of regenerated plants was successfully achieved by planting them in peat moss substrate after a desiccation treatment at room temperature for 3 d.     

Further investigation of the mechanism of <Emphasis Type=Italic>Vitreoscilla</Emphasis> hemoglobin (VHb) protection from oxidative stress in <Emphasis Type=Italic>Escherichia coli</Emphasis>

In Escherichia coli, Vitreoscilla hemoglobin (VHb) protects against oxidative stress, perhaps, in part, by oxidizing OxyR. Here this protection, specifically VHb-associated effects on superoxide dismutase (SOD) and catalase levels, was examined. Exponential or stationary phase cultures of SOD⁺ or SOD⁻ E. coli strains with or without VHb and oxyR antisense were treated with 2 mM hydrogen peroxide without sublethal peroxide induction, and compared to untreated control cultures. The hydrogen peroxide treatment was toxic to both SOD⁺ and SOD⁻ cells, but much more to SOD⁻ cells; expression of VHb in SOD⁺ strains enhanced this toxicity. In contrast, the presence of VHb was generally associated in the SOD⁺ background with a modest increase in SOD activity that was not greatly affected by oxyR antisense or peroxide treatment. In both SOD⁺ and SOD⁻ backgrounds, VHb was associated with higher catalase activity both in the presence and absence of peroxide. Contrary to its stimulatory effects in stationary phase, in exponential phase oxyR antisense generally decreased VHb levels.     

Micromorphological differences between some European and American <Emphasis Type=Italic>Fraxinus</Emphasis> (<Emphasis Type=Italic>Oleaceae</Emphasis>) species

Micromorphological differences in leaves and pollen between two American (Fraxinus americana L., F. pennsylvanica Marshall) and two European (F. angustifolia Vahl, F. excelsior L.) ash species were studied using scanning electron microscope. The types, dimensions and distribution of characteristic trichomes were established and measured. Capitate hairs on the leaves had the same shape in all researched ash species. Acicular hairs were regularly present in two American ash species, but very rarely in the glabrous phase of F. angustifolia and F. excelsior. Only F. americana had coronulate abaxial surface of leaves. Pollen of F. angustifolia and F. excelsior had 3 (tricolpate) apertures, and F. americana and F. pennsylvanica 4 (stephanocolpate) apertures. Based on the appearance of the reticulum it’s possible to clearly distinguish all four species. F. angustifolia and F. pennsylvanica had muri with transversal ridges and seldom granules. Muri of F. excelsior and F. americana had slightly visible transversal ridges, and because of that noticeable granules.     

<Emphasis Type=Italic>Gagea robusta</Emphasis> (Liliaceae), a new species from <Emphasis Type=Italic>Flora Iranica</Emphasis> area

A new species, endemic to the Flora Iranica area, Gagea robusta (Liliaceae) is described and illustrated. It differs from G. reticulata (Pall.) Schult. & Schult. f. in having a puberulent indumentum on the basal leaf and from G. setifolia Baker ex Aitch. in having a long neck (up to 12 cm long). Its ecology, distribution and conservation status are discussed.