<Emphasis Type="Italic">Eschweilera awaensis</Emphasis> and <Emphasis Type="Italic">Grias subbullata</Emphasis> (Lecythidaceae), two new species from northwestern Ecuador

Eschweilera awaensis and Grias subbullata, two new species known from the wet forests of northwestern Ecuador, are described. Eschweilera awaensis occurs in Esmeraldas, Los Ríos, and the westernmost parts of the Pichincha provinces between 25–650 m elevation. Grias subbullata, is known only from two localities in the Santo Domingo de los Tsáchilas province between 900–1670 m elevation. The new species are illustrated and their relationships with similar species are discussed.     

<Emphasis Type="Italic">Capparidastrum tuxtlense</Emphasis> (Capparaceae), a new Mexican endemic

We describe and illustrate Capparidastrum tuxtlense, a narrow endemic, known only from the eastern slopes of Volcán San Martín, (10–450 m), Veracruz, Mexico. Capparidastrum tuxtlense is similar to Capparidastrum discolor, but differs by foliar, floral, and fruit characters. A discussion of putative relatives and a key to the Central American species of Capparidastrum are included.     

<Emphasis Type="Italic">Grias theobromicarpa</Emphasis> (Lecythidaceae), a new species from northwestern Ecuador

Grias theobromicarpa, a new species from northwestern Ecuador is described. This species is found in wet forests at ca. 1500 m elevation, and is distinguished by its erect petals that conceal the androecium, filaments fused to form a very distinct staminal tube, anthers that are oblong with cucullate connectives enclosing most of the thecae, and especially by its fruits, which are prominently ridged when fresh. The species is illustrated and its relationship with its closest relative, G. purpuripetala, from southwestern Colombia, is discussed.     

Development of a real-time TaqMan assay to detect <Emphasis Type="Italic">mendocina</Emphasis> sublineage <Emphasis Type="Italic">Pseudomonas</Emphasis> species in contaminated metalworking fluids

A TaqMan quantitative real-time polymerase chain reaction (qPCR) assay was developed for the detection and enumeration of three Pseudomonas species belonging to the mendocina sublineage (P. oleovorans, P. pseudoalcaligenes, and P. oleovorans subsp. lubricantis) found in contaminated metalworking fluids (MWFs). These microbes are the primary colonizers and serve as indicator organisms of biodegradation of used MWFs. Molecular techniques such as qPCR are preferred for the detection of these microbes since they grow poorly on typical growth media such as R2A agar and Pseudomonas isolation agar (PIA). Traditional culturing techniques not only underestimate the actual distribution of these bacteria but are also time-consuming. The primer–probe pair developed from gyrase B (gyrB) sequences of the targeted bacteria was highly sensitive and specific for the three species. qPCR was performed with both whole cell and genomic DNA to confirm the specificity and sensitivity of the assay. The sensitivity of the assay was 10¹ colony forming units (CFU)/ml for whole cell and 13.7 fg with genomic DNA. The primer–probe pair was successful in determining concentrations from used MWF samples, indicating levels between 2.9 × 10³ and 3.9 × 10⁶ CFU/ml. In contrast, the total count of Pseudomonas sp. recovered on PIA was in the range of <1.0 × 10¹ to 1.4 × 10⁵ CFU/ml for the same samples. Based on these results from the qPCR assay, the designed TaqMan primer–probe pair can be efficiently used for rapid (within 2 h) determination of the distribution of these species of Pseudomonas in contaminated MWFs.     

<Emphasis Type="Italic">Tomentella furcata</Emphasis>, a new species from Benin (West Africa) with basidia forming internal hyphae

In this study, a new tropical African resupinate thelephoroid species, namely Tomentella furcata, is described. Microscopically, it is characterised by mostly simple septate and thick-walled (1–2 μm) subicular and subhymenial hyphae, suburniform basidia with transverse septa and/or infrequently with an internal hypha, large sterigmata of 2–4 μm at base, and distinctly yellow, subglobose to globose basidiospores (both in frontal and lateral view) with long (1–2 μm) and dense, simple or forked spines. Phylogenetically, T. furcata falls within terminal clades of investigated Tomentella species. We discuss ornamentation types of basidiospores for discrimination of resupinate thelepohoroid genera and confirm the limitation of ornament bifurcation as a diagnostic feature of Pseudotomentella. Anatomical dissimilarities between T. furcata and close species are given. Taxonomical novelty: Tomentella furcata Yorou & Agerer.     

Water relations advantages for invasive <Emphasis Type="Italic">Rubus</Emphasis> <Emphasis Type="Italic">armeniacus</Emphasis> over two native ruderal congeners

Despite species in the Rubus fruticosus complex (wild blackberry) being among the most invasive plants globally in regions with large annual fluctuations in water availability, little is known about their water relations. We compared water relations of a prominent member of the complex, R. armeniacus (Himalayan blackberry), with species native to the Pacific Northwest of North America (PNW), R. spectabilis (salmonberry) and R. parviflorus (thimbleberry). In eight stands of each species located near Portland, Oregon, USA, we measured mid-day hydraulic resistance (R _plant), and daily time series of stomatal conductance (g _s), leaf water potential (Ψ_lf), and environmental conditions at four time periods spanning the 2007 growing season. Although all species maintained Ψ_lf above −0.5 MPa in spring, R. armeniacus maintained less negative Ψ_lf (≥−1.0 MPa) than the natives in summer, a factor attributable to advantages in both its root and shoot systems. R _plant of R. armeniacus was ≤0.1 MPa mmol⁻¹ m² s for the duration of the study, and approximately 25–50% of R _plant for the native species in summer. R. armeniacus had higher g _s compared to the native species throughout the spring and summer, with approximately twice their rates in summer. Our R _plant and g _s results show that R. armeniacus has access to more water during PNW summers than congeneric natives, allowing it to maintain high water-use, and potentially helping it achieve higher growth and reproductive rates. Water relations may therefore be a critical component of the competitive and invasive success of R. armeniacus and other R. fruticosus species worldwide.     

A new armored searobin <Emphasis Type="Italic">Paraheminodus longirostralis</Emphasis> (Teleostei: Peristediidae) from New Caledonia

A new species of armored searobin, Paraheminodus longirostralis, is described from five specimens collected from New Caledonia at depths of 412–467 m. It is distinguishable from its three known congeners in having 34 bony plates in the upper lateral row, a forward-directed spine on each plate between the 23rd–26th and 31st–32nd plates in the upper lateral row, 6–7 + 1 + 20–21 = 27–28 gill rakers, an elongate body posterior to the anus (49.9–52.1% standard length), an elongate rostral projection (53.0–59.3% head length), short upper jaw (42.1–43.4% head length), an elongate pectoral fin (70.6–79.4% head length), and long preopercular spine (39.2–57.7% head length).     

A new species of <Emphasis Type="Italic">Lonchocarpus</Emphasis> (Leguminosae,Papilionoideae) from Mato Grosso do Sul,Brazil

Lonchocarpus variabilis, from Mato Grosso do Sul State in Midwestern Brazil, is newly described and illustrated on the basis of field and herbarium studies. It occurs in seasonally deciduous forests and gallery forests at 150–681 m elevation. This species grows in sandy, calcareous, and iron rocky soils. It is similar to L. fendleri, but differs in number of leaflets and shape of the calyx. Moreover, L. variabilis has a staminal tube dilated at the base and anthers with a glabrous base.     

Dos especies nuevas de <Emphasis Type="Italic">Borreria</Emphasis> (Rubiaceae), un nuevo registro de <Emphasis Type="Italic">Declieuxia</Emphasis> y observaciones taxonómicas

Two new species of Borreria are described: the first, Borreria johnwoodii from the Campo Cerrado area of the Noel Kempff Mercado National Park; the second, Borreria siberiana from the cloud forest of the Yungas, between 2600 and 3100 m above sea level. Declieuxia cordigera is recorded for the first time from Bolivia. Borreria nectarifera is shown to be synonymous with B. eryngioides var. affinis.     

Reclassification of Two <Emphasis Type="Italic">Peronospora</Emphasis> Species Parasitic on <Emphasis Type="Italic">Draba</Emphasis> in <Emphasis Type="Italic">Hyaloperonospora</Emphasis> Based on Morphological and Molecular Phylogenetic Data

On the family Brassicaceae, the causal agent responsible for downy mildew disease was originally regarded as a single species, Peronospora parasitica (now under Hyaloperonospora), but it was recently reconsidered to consist of many distinct species. In this study, 11 specimens of Peronospora drabae and P. norvegica parasitic on the genus Draba were investigated morphologically and molecularly. Pronounced differences in conidial sizes (P. drabae: 14–20 × 12.5–15.5 μm; P. norvegica: 20–29 × 15.5–22 μm) and 7.8% sequence distance between their ITS1-5.8S-ITS2 rDNA sequences confirmed their status as distinct species. Based on ITS phylogeny and morphology (monopodially branching conidiophores, flexuous to sigmoid ultimate branchlets, hyaline conidia and lobate haustoria), the two species unequivocally belong to the genus Hyaloperonospora and not to Peronospora to which they were previously assigned. Therefore, two new combinations, Hyaloperonospora drabae and H. norvegica, are proposed. The two taxa are illustrated and compared using the type specimen for H. norvegica and authentic specimens for H. drabae, which is lectotypified.     

Conversion of soybean sterols into 3,17-diketosteroids using actinobacteria <Emphasis Type="Italic">Mycobacterium neoaurum,Pimelobacter simplex</Emphasis>, and <Emphasis Type="Italic">Rhodococcus erythropolis</Emphasis>

Soybean sterols were converted into androst-4-ene-3,17-dione (AD) and 9α-hydroxyandrost-4-ene-3,17-dione (9-OH-AD) using three actinobacterium strains. The transformation of a microcrystallic substrate (particle size 5–15 μm) or the transformation in the presence of randomly methylated β-cyclodextrin (MCD) were carried out by Mycobacterium neoaurum with a phytosterol load of 30 g/l over 144 h with an AD content of 14.5 and 15.2 g/l, respectively. AD obtained in the presence of MCD was transformed into ADD (13.5 g/l) by Pimelobacter simplex cells over 3 h and into 9-OH-AD by Rhodococcus erythropolis cells after 22 h without the isolation of AD from the cultural liquid. The crude product ADD was obtained in 75% yield, based on phytosterol. It contained as by-products 1.25% of AD and 1.5% of 1,2-dehydrotestosterone. In a control experiment—the process of 1,2-dehydrogenation of 20 g/l AD in the water solution of MCD—no by-products were isolated. Thus, it is more expedient to introduce the 1,2-double bond into pure AD, whereas R. erythropolis strain with low destructive activity towards steroid nucleus can be used in the mixed culture with M. neoaurum. The crystal product contained, according to HPLC, 80% of 9-OH-AD, and 1.5% AD was obtained. The yield of 9-OH-AD (m.p. 218–220°C) based on transformed phytosterol was 56%.     

Utilization of dibenzothiophene as sulfur source by <Emphasis Type="Italic">Microbacterium</Emphasis> sp. <Emphasis Type="Italic">NISOC-06</Emphasis>

Oil-polluted soils were sampled from National Iranian South Oil Company (NISOC) for isolation and screening of C–S and not C–C targeted Dibenzothiophene (DBT) degrading microorganisms. Microbacterium sp. NISOC-06, a C–S targeted DBT degrading bacterium, was selected and its desulfurization ability was studied in aqueous phase and water-gasoline biphasic systems. The 16srRNA gene was amplified using universal eubacteria-specific primers, PCR product was sequenced and the sequence of nearly 1,500 bp 16srDNA was studied. Based on Gas Chromatography results Microbacterium sp. NISOC-06 utilized 94.8% of 1 mM DBT during the 2 weeks of incubation. UV Spectrophotometry and biomass production measurements showed that the Microbacterium sp. NISOC-06 was not able to utilize DBT as a carbon source. There was no accumulation of phenolic compounds as Gibb’s assay showed. Biomass production in a biphasic system for which DBT-enriched gasoline was used as the sulfur source indicated the capability of Microbacterium sp. NISOC-06 to desulfurize gasoline.     

Chemical composition of the essential oil from two wormwood species <Emphasis Type="Italic">Artemisia frigida</Emphasis> and <Emphasis Type="Italic">Artemisia argyrophylla</Emphasis>

The composition of the essential oil from the wormwood sage (Artemisia frigida Willd., Asteraceae) of populations growing in the Altai Territory, the Altai Republic, the Khakass Republic, the Tuva Republic, and the East-Kazakhstan region of the Republic of Kazakhstan and the representative species of the silver-leaved wormwood Artemisia argyrophylla Ledeb. growing in the Republic Altai has been studied by chromato-mass spectrometry. An analysis of 15 samples of the essential oil from A. frigida obtained over a period from 1999 to 2007 indicates that samples from different populations have similar sets of the main components: α-pinene (0.2–7.8%), camphene (1.9–5.8%), 1,8-cineole (8.9–33.8%), camphor (6.7–40.0%), borneol (3.9–12.3%), terpine-4-ol (1.5–6.5%), bornyl acetate (1.4–22.0%), and germacrene D (1.4–14.6%). Some samples contain substantial amounts of α- and β-thujones (in total up to 19.1%), which are completely absent in other samples. Some samples contain santolina alcohol (up to 13.8%) and its acetate (up to 4.8%). As differentiated from A. frigida, the essential oil of A. argyrophylla contains yomogi alcohol (1.2%), artemisia ketone (12.9%), artemisia alcohol (3.1%), artemisia alcohol acetate (3.9%), and small amounts of camphor (3.2%), borneol (0.3%), and bornyl acetate (0.2%).     

Direct gene transfer study and transgenic plant regeneration after electroporation into mesophyll protoplasts of <Emphasis Type="Italic">Pelargonium</Emphasis> <Emphasis Type="Italic">×</Emphasis> <Emphasis Type="Italic">hortorum</Emphasis>, ‘Panaché Sud’

Direct genetic transformation of mesophyll protoplasts was studied in Pelargonium × hortorum. Calcein and green-fluorescent protein (GFP) gene were used to set up the process. Electroporation (three electric pulses from a 33-μF capacitor in a 250-V cm⁻¹ electric field) was more efficient than PEG 6000 for membrane permeation, protoplast survival and cell division. Transient expression of GFP was detected in 33–36% of electroporated protoplasts after 2 days and further in colonies. A protoplast suspension conductivity of >1,500 μS cm⁻¹ allowed high colony formation and plant regeneration. Stable transformation was obtained using the plasmid FAJ3000 containing uidA and nptII genes. When selection (50 mg l⁻¹ kanamycin) was achieved 6 weeks after electroporation, regenerated shoots were able to grow and root on 100 mg l⁻¹ kanamycin. The maximum transformation efficiency was 4.5%, based on the number of colonies producing kanamycin-resistant rooted plants or 0.7% based on the number of cultured protoplasts. Polymerase chain reaction (PCR) analysis on in vitro micropropagated plants showed that 18 clones out of 20 contained the nptII gene, while the uidA gene was absent. These results were confirmed after PCR analyses of five glasshouse-acclimatized clones.     

Isolation of a <Emphasis Type="Italic">Ve</Emphasis> homolog, <Emphasis Type="Italic">mVe1</Emphasis>, and its relationship to verticillium wilt resistance in <Emphasis Type="Italic">Mentha longifolia</Emphasis> (L.) Huds

As a step toward greater understanding of the genetics of verticillium wilt resistance in plants, we report the sequencing of a candidate wilt resistance gene, mVe1, from the mint diploid model species, Mentha longifolia (Lamiaceae). mVe1 is a putative homolog of tomato (Solanum lycopersicum L.) verticillium wilt (Ve) resistance genes. The mVe1 gene has a coding region of 3,051 bp. The predicted mVe1 protein contains a leucine-rich repeat domain, a common feature of plant disease resistance proteins. We compared 13 mVe1 alleles from three mint species. These alleles shared 96.2–99.6% nucleotide identity. We analyzed four M. longifolia populations segregating with respect to mVe1 alleles and wilt resistance versus susceptibility and found one association between mVe1 genotype and wilt phenotype. We conclude that mVe1 may play a role in mint verticillium wilt resistance, but variation for resistance in our segregating progenies is likely polygenic. Therefore, further investigations of mVe1 and identification of additional candidate genes are both warranted.     

Synthesis of orthogonally protected (<Emphasis Type="Italic">3R</Emphasis>,<Emphasis Type="Italic">4S</Emphasis>)- and (<Emphasis Type="Italic">3S</Emphasis>,<Emphasis Type="Italic">4S</Emphasis>)-4,5-diamino-3-hydroxypentanoic acids

Summary.  The paper describes two methods of the synthesis of ethyl (3R,4S)- and (3S,4S)-4-[(benzyloxycarbonyl)amino]-5-[(tert-butyloxycarbonyl)amino]-3-hydroxypentanoates, useful for the syntheses of edeine analogs. Differently N-protected (S)-2,3-diaminopropanoic acid was used as a substrate in both procedures. The absolute configuration of newly generated asymmetric carbon atoms C-3 in β-hydroxy-γ,δ-diamino products was assigned by means of ¹H NMR spectroscopy after their transformation into corresponding piperidin-2-ones. Received May 24, 2002 Accepted October 10, 2002 Published online December 18, 2002 Acknowledgment The authors are indebted to the Faculty of Chemistry, Technical University of Gdańsk for financial support. Authors' address: Zbigniew Czajgucki, M. Sc., Department of Pharmaceutical Technology and Biochemistry, Faculty of Chemistry, Technical University of Gdańsk, 11/12 Narutowicza St., 80-952 Gdańsk, Poland, Fax +48 58 347 11 44, E-mail: zmczaj@wp.pl     

Cell lines from the soft tick <Emphasis Type="Italic">Ornithodoros moubata</Emphasis>

Primary cell cultures (n = 16) were initiated from tissues of embryonic and neonatal larval Ornithodoros moubata following methods developed for hard ticks. After maintenance for 20–25 months in vitro, cell multiplication commenced in surviving cultures, leading to the establishment of six cell lines designated OME/CTVM21, 22, 24, 25, 26 and 27. All lines are maintained at 28°C, with subculture at 2–8 week intervals. The cultures comprise heterogeneous populations of large cells of 15–100 μm in diameter, often with finger-like protrusions and/or intracellular crystals, rarely attached, predominantly floating and forming clumps or hollow multicellular vesicles up to 1 mm in diameter. Attempts to cryopreserve the cells are described. Tick-borne encephalitis virus has been serially passaged ten times in OME/CTVM21 cells without significant decrease in virus production and with no change in its biological properties as shown by the size and morphology of plaques produced in porcine kidney cells.     

Exploring the potential of clay in mitigating <Emphasis Type="Italic">Pyrodinium bahamense</Emphasis> var. <Emphasis Type="Italic">compressum</Emphasis> and other harmful algal species in the Philippines

Harmful algal bloom occurrences worldwide have prompted the testing and use of methods to control and mitigate their detrimental effects. This study investigates the potential of Philippine clay minerals to physically remove phytoplankton cells under laboratory conditions. Ball clay had the highest removal efficiency (∼95%) for Pyrodinium bahamense (paralytic shellfish poisoning causative organism) cells. A slight decrease in the efficiency by 10–20% was seen when culture volume was increased from 50 mL to 1 L. Removal efficiency was reduced to ∼95% when water motion was introduced. Removal of other phytoplankton species (Gymnodinium sanguineum, Amphidinium carterae, Pyrophacus horologium, Chatonella marina, and Alexandrium sp.) using ball clay was less efficient (<70%). Cell removal efficiencies differed with phytoplankton species belonging to the same taxonomic group. Possible mechanisms for cell removal are described.