Corynebacterium antarcticum sp. nov., Corynebacterium marambiense sp. nov., Corynebacterium meridianum sp. nov., and Corynebacterium pygosceleis sp. nov., isolated from Adélie penguins (Pygoscelis adeliae)

A taxonomic study was conducted on 16 bacterial strains isolated from wild Adélie penguins (Pygoscelis adeliae) from Seymour (Marambio) Island and James Ross Island. An initial screening by repetitive sequence-based PCR fingerprinting divided the strains studied into four coherent groups. Phylogenetic analysis based on 16S rRNA gene sequences assigned all groups to the genus Corynebacterium and showed that Corynebacterium glyciniphilum and Corynebacterium terpenotabidum were the closest species with 16S rRNA gene sequence similarities between 95.4 % and 96.5 %. Further examination of the strains studied with ribotyping, MALDI-TOF mass spectrometry, comprehensive biotyping and calculation of average nucleotide identity and digital DNA–DNA hybridisation values confirmed the separation of the four groups from each other and from the other Corynebacterium species. Chemotaxonomically, the four strains P5828^T, P5850^T, P6136^T, P7210^T representing the studied groups were characterised by C_16:0 and C_18:1 ω9c as the major fatty acids, by the presence of meso-diaminopimelic acid in the peptidoglycan, the presence of corynemycolic acids and a quinone system with the predominant menaquinone MK-9(H₂). The results of this study show that the strains studied represent four new species of the genus Corynebacterium, for which the names Corynebacterium antarcticum sp. nov. (type strain P5850^T = CCM 8835^T = LMG 30620^T), Corynebacterium marambiense sp. nov. (type strain P5828^T = CCM 8864^T = LMG 31626^T), Corynebacterium meridianum sp. nov. (type strain P6136^T = CCM 8863^T = LMG 31628^T) and Corynebacterium pygosceleis sp. nov. (type strain P7210^T = CCM 8836^T = LMG 30621^T) are proposed.     

Germplasm conservation of the tropical forest trees,Cedrela odorata L.,Guazuma crinita Mart., andJacaranda mimosaefolia D. Don., by shoot tip encapsulation in calcium-alginate and storage at 12–25°C

Germplasm conservation of the tropical forest trees,Cedrela odorata L.,Guazuma crinita Mart., andJacaranda mimosaefolia D. Don., at above-freezing temperatures following alginate-bead encapsulation was attempted. Shoot tips excised from in vitro plantlets were encapsulated in calcium-alginate beads and stored on different substrates at 12, 20, and 25 °C. Percent viability when encapsulated shoot tips were stored on substrate containing only water solidified with 1% (wt/vol) agar was 80% after 12 months at 12°C forC. odorata, 90% after 12 months at 25°C forG. crinita, and 70% after 6 months at 20°C forJ. mimosaefolia.Abbreviations BAP 6-Benzylaminopurine - KIN 6-Furfurylaminopurine     

Morphology and cell division of the oxytrichids Architricha indica nov. gen., nov. sp., and Histriculus histrio (Müller, 1773), Corliss, 1960 (Ciliophora, Hypotrichida)

The oxytrichid ciliate Architricha indica nov. gen., nov. sp., isolated from the river Yamuna, Delhi, shows a new combination of characters. It possesses a flexible body, 18 frontal-ventral-transverse (FVT) cirri, 3 right and 2 left marginal cirral rows, 6 dorsal bristle rows and 3 caudal cirri (CC). The FVT cirri arise from 6 primordia, which utilize 6 parental cirri in their origin as is typical of Oxytricha species. Multiple marginal rows (MMR) develop through 5 independent marginal primordia arising "within-row", 1 in each parental marginal row. All the 5 marginal rows are thus morphogenetically active. Such a mode of formation of MMR has not been recorded among oxytrichids and has necessitated separation of A. indica at the generic level. Histriculus, on the other hand, has well-known characteristics, viz. rigid body, confluent marginal rows and absence of CC. The morphogenesis of Histriculus histrio has been described by Berger and Foissner [1997. Cladistic relationships and generic characterization of oxytrichid hypotrichs (Protozoa, Ciliophora). Arch. Protistenkd. 148, 125-155]. Reinvestigation of very early stages of development revealed that (i) the FVT cirral primordia utilize kinetosomes from 5 parental FVT cirri, (ii) the primordium II of the proter is of a composite origin: kinetosomes from the oral primordium merge with the primordium II that originates from the buccal cirrus II/2 and (iii) the FVT primordia V and VI for the 2 daughter cells arise sequentially from the parental cirrus V/4. Thus, the genus Histriculus exhibits a new combination of characters with respect to the origin of FVT cirri, an additional pattern to be added to the known 6 patterns of FVT development in oxytrichids [Berger and Foissner, 1997; Berger, H., 1999. Monograph of the Oxytrichidae (Ciliophora, Hypotrichida), Kluwer Academic Publishers, Dordrecht/Boston/London].     

Seed structure in Crocus sativus L. ×, C. cartwrightianus Herb., C. thomasii Ten., and C. hadriaticus Herb. at SEM

Crocus sativus L. (Iridaceae) is a sterile triploid geophyte widely cultivated for the production of the spice saffron and only reproduced by means of corms. Extensive research has identified Crocus cartwrightianus Herb. as being a probable progenitor of C. sativus. However, other diploid Crocus species of the same C. sativus group, such as C. thomasii Ten. and C. hadriaticus Herb., have been considered as possible progenitors of saffron. Of the characteristics for distinguishing critical genera, species and intraspecific taxa of angiosperms, the most widely adopted have been seed organisation and patterns of spermoderma microstructure detected at SEM. The aim of this study is to use SEM to analyse the seeds of C. sativus ×, a cross obtained by C. sativus with pollen of C. cartwrightianus Herb. and the seeds of allopollinated C. cartwrightianus, C. thomasii Ten., and C. hadriaticus Herb. Results indicate that the seed surface microstructure of C. sativus × is very similar to that of C. cartwrightianus while being different from those of C. thomasii and C. hadriaticus.     

Seed banks of managed and degraded grasslands in the Krkonoše Mts., Czech Republic

Grassland seed banks are traditionally considered a source of new species in degraded communities. However, many recent studies have shown that the potential of the seed bank to restore many communities is rather limited. Two principal reasons for these limitations, loss of species from the seed bank or inability of the species to create any seed bank, are, however, usually not distinguished. This study aims to assess the role of seed bank composition and heterogeneity in the restoration of species-rich plant communities. It was carried out in mountain grasslands in the eastern part of the Krkono?e Mountains, Czech Republic. The composition of vegetation and seed bank were recorded and their relationship was assessedin 1.5m × 1.5 m plots placed in non-degraded and degraded parts of seven grasslands. Vegetation at currently managed sites is not degraded; degraded parts were without management (dominated byHolcus mollis, Bistorta major orRumex alpinus). The degree of heterogeneity of seed bank and vegetation was tested as the relationship between subplot similarity, distance, and degradation stage. Degradation had significant effects on composition of both aboveground vegetation and seed bank and increased heterogeneity both in the vegetation and in the seed bank. Species absent from the vegetation of degraded plots were also absent from the seed bank of both degraded and non-degraded plots, indicating that the absence of species from the seed bank is not due to their loss during the degradation process but rather due to the low number of seeds in the seed bank already in the non-degraded communities. Furthermore, the seed bank of the degraded communities largely results from the present vegetation of these communities. This supports the limited role of the seed bank in these communities. Restoration of these sites is thus impossible unless management will include methods with which seeds will arrive at the degraded sites.     

Phenotypic and genotypic analysis of clarithromycin-resistant Helicobacter pylori from Bogotá D.C., Colombia

Resistance of Helicobacter pylori to clarithromycin is the most common cause of treatment failure in patients with H. pylori infections. This study describes the MICs and the presence of 23S rRNA mutations of H. pylori isolates from Bogotá, D.C., Colombia. H. pylori were isolated from gastric biopsies from patients with functional dyspepsia. Clarithromycin susceptibility was investigated by agar dilution and strains were considered resistant if the MIC was ≥1 μg/ml. DNA sequences of the 23S rRNA gene of strains resistant and sensitive to clarithromycin were determined to identify specific point mutations. Clarithromycin resistance was present in 13.6% of patients by agar dilution. The A2143G, A2142G and A2142C mutations were found in 90.5, 7.1, and 2.4% of H. pylori strains with resistance genotype.The resistant phenotype was associated with 23S rRNA resistance genotype in 85.7% of isolates. The point mutations in 23S rRNA were well correlated with MICs values for clarithromycin.     

Neokomagataea gen. nov., with descriptions of Neokomagataea thailandica sp. nov. and Neokomagataea tanensis sp. nov., osmotolerant acetic acid bacteria of the α-Proteobacteria

Isolates AH11(T) and AH13(T) were isolated from flowers of lantana and candle bush respectively collected in Thailand. In phylogenetic trees based on 16S rRNA gene sequences, the two isolates formed an independent cluster, which was then connected to the type strain of Saccharibacter floricola. The calculated pair-wise 16S rRNA gene sequence similarities of isolate AH11(T) were 95.7-92.3% to the type strains of the type species of the 12 genera of acetic acid bacteria. The DNA base composition was from 51.2 to 56.8 mol % G+C, with a range of 5.6 mol %. When isolate AH11(T) was labeled, DNA-DNA similarities were 100, 12, 4, 5, and 4% respectively to isolates AH11(T) and AH13(T) and the type strains of Saccharibacter floricola, Gluconobacter oxydans, and Acetobacter aceti. The two isolates were non-motile and did not oxidize either acetate or lactate. No growth was found in the presence of 0.35% acetic acid w/v. The two isolates were not osmophilic but osmotolerant, produced 2,5-diketo-D-gluconate from D-glucose, and did not oxidize lactate, thus differing from strains of Saccharibacter floricola, which showed weak lactate oxidation. The two isolates contained unsaturated C(18:1)ω7c fatty acid as the major fatty acid, and were unique in the presence of a considerable amount of straight-chain C(18:1)2OH fatty acid. Q-10 was present as the major isoprenoid quinone. Neokomagataea gen. nov. was proposed with the two species, Neokomagataea thailandica sp. nov. for isolate AH11(T) (=BCC 25710(T)=NBRC 106555(T)), which has 56.8 mol % G+C, and Neokomagataea tanensis sp. nov. for isolate AH13(T) (=BCC 25711(T)=NBRC 106556(T)), which has 51.2 mol % G+C.     

Neokomagataea gen. nov., with Descriptions of Neokomagataea thailandica sp. nov. and Neokomagataea tanensis sp. nov., Osmotolerant Acetic Acid Bacteria of the α-Proteobacteria

Isolates AH11^T and AH13^T were isolated from flowers of lantana and candle bush respectively collected in Thailand. In phylogenetic trees based on 16S rRNA gene sequences, the two isolates formed an independent cluster, which was then connected to the type strain of Saccharibacter floricola. The calculated pair-wise 16S rRNA gene sequence similarities of isolate AH11^T were 95.7–92.3% to the type strains of the type species of the 12 genera of acetic acid bacteria. The DNA base composition was from 51.2 to 56.8 mol % G+C, with a range of 5.6 mol %. When isolate AH11^T was labeled, DNA-DNA similarities were 100, 12, 4, 5, and 4% respectively to isolates AH11^T and AH13^T and the type strains of Saccharibacter floricola, Gluconobacter oxydans, and Acetobacter aceti. The two isolates were non-motile and did not oxidize either acetate or lactate. No growth was found in the presence of 0.35% acetic acid w/v. The two isolates were not osmophilic but osmotolerant, produced 2,5-diketo-D-gluconate from D-glucose, and did not oxidize lactate, thus differing from strains of Saccharibacter floricola, which showed weak lactate oxidation. The two isolates contained unsaturated C_18:1ω7c fatty acid as the major fatty acid, and were unique in the presence of a considerable amount of straight-chain C_18:12OH fatty acid. Q-10 was present as the major isoprenoid quinone. Neokomagataea gen. nov. was proposed with the two species, Neokomagataea thailandica sp. nov. for isolate AH11^T (=BCC 25710 ^T =NBRC 106555^T), which has 56.8 mol % G+C, and Neokomagataea tanensis sp. nov. for isolate AH13^T (=BCC 25711^T=NBRC 106556^T), which has 51.2 mol % G+C.     

Cryopreservation of Cymbidium eburneum Lindl. and C. hookerianum Rchb. f., two threatened and vulnerable orchids via encapsulation–dehydration

A successful cryopreservation protocol for the long-term conservation of protocorms of two threatened and vulnerable orchids, Cymbidium eburneum Lindl. and Cymbidium hookerianum Rchb. f., was developed using encapsulation–dehydration. Protocorms were osmoprotected in liquid Murashige and Skoog medium (MS) containing 0.7 M sucrose for 20 h at 25?±?2°C on a rotary shaker, and incorporated into an encapsulation matrix [consisting of 3% (w/v) sodium alginate and 100 mM CaCl₂]. The encapsulated protocorms, which were desiccated in a laminar airflow cabinet for 6 h, were able to withstand cryostorage in liquid nitrogen. Maximum regeneration into complete plantlets (72% for C. eburneum and 70% for C. hookerianum) of the cryostored, encapsulated protocorms was obtained using MS medium containing 3% sucrose and 0.8% agar. Using this protocol of cryopreservation, long-term preservation for ex situ conservation of these two threatened orchids can be accomplished.