Novel Aerobic Methylotrophic Isolates from the Soda Lakes of the Southern Transbaikal Region

Twenty-one bacterial associations isolated from the soda lakes of the southern Transbaikal region were found to be able to actively grow at pH 9–10 on methanol as the source of carbon and energy. Two alkalitolerant facultatively methylotrophic strains, Bur 3 and Bur 5, were obtained in pure cultures. Both strains represent gram-negative, nonmotile, bean-shaped, encapsulated cells that reproduce by binary fission. The strains are able to grow at temperatures ranging from 6 to 42°C, with an optimum growth temperature of 25–29°C (strain Bur 3) and 35–37°C (strain Bur 5) and at pH between 6.5 and 9.5, with an optimum pH value of 8.0–8.5. At pH 9.0, strain Bur 3 exhibits an increased content of phosphatidylglycerol and a decreased content of phosphatidylethanolamine. Strains Bur 3 and Bur 5 are similar in the G+C content of their DNAs (66.2 and 65.5 mol %, respectively) and in the type of the dominant ubiquinone (Q ₁₀). Unlike Bur 5, strain Bur 3 is able to grow autotrophically in an atmosphere of CO₂+ O₂+ H₂. The strains oxidize, by the respective dehydrogenases, methanol to CO₂, which is assimilated by the ribulose bisphosphate pathway. Ammonium ions are assimilated in the glutamate cycle and by the reductive amination of -ketoglutarate. The strains are highly homologous to each other (92%) and are much less homologous (at a level of 28–35%) to representatives of the genus Ancylobacter, A. aquaticusATCC 25396^Tand A. vacuolatumDSM 1277. Based on the results obtained, both strains are assigned to a new species, Ancylobacter natronumsp. nov.     

A new nitrogen-fixing bacterium: Derxia gummosa nov. gen. nov. spec.

Summary A vigorously nitrogen-fixing bacterium was isolated from a West Bengal soil. Cells from young cultures on nitrogen-deficient agar medium appear as rods measuring approx. 3–6x1.0–1.2 ., with rounded ends and homogeneous cytoplasm. Older cells contain big refractile bodies representing vacuoles and fat deposits and show a wide variation in shape. Short motile cells with a single polar flagellum arise in liquid medium with combined (ammonia) nitrogen. The Gram reation is negative and endospores are not formed.The organism is aerobic but catalase-negative. It grows rather feebly in sugar-free protein media, but in nitrogen-deficient media it produces ah heavy and extremely tenacious growth, of a characteristic uneven appearance on agar and changing from white to dark red-brown. Glucose, fructose, ethanol, glycerol, mannitol and sorbitol are the best sources of carbon. Acid but no gas is formed from several carbohydrates. Nitrogen fixation is slow but may amount to 20–25 mg. per gram glucose or mannitol supplied. Molybdenum stimulates nitrogen fixation strongly and cannot be replaced by vanadium.Various forms of combined nitrogen, especially glutamic acid, are assimilated more readily than free nitrogen. Growth can be initiated within aPh-interval from approx. 5 to 9; growing cultures may be acidified to Ph 4.5 or less.The organism is morphologically, culturally and serologically different from Azotobacter and Beijerinckia and is regarded as representing a new genus.     

A new facultatively nitrite oxidizing bacterium,Nitrobacter vulgaris sp. nov.

A total of 17 facultatively lithoautotrophic strains of Nitrobacter were investigated. They all were found to be related on the species level by DNA hybridizations. The G+C content of DNA ranged between 58.9 and 59.9 mol %. The isolates originated from divers environments. The cells were 0.5–0.8×1.2–2.0 m in size and motile by one polar to subpolar flagellum. Cell-division normally occurred by budding. Polar caps of intracytoplasmic membranes as well as carboxysomes were present. The cells tended to excrete extracellular polymers forming aggregates or biofilms. Heterotrophic growth was slower than mixotrophic but often faster than litoautotrophic growth. In the presence of nitrite and organic substances the organisms often showed diphasic growth. First nitrite and then the organic material was oxidized. In the absence of oxygen growth was possible by dissimilatory nitrate reduction. Nitrite, nitric and nitrous oxide as well as ammonia were formed. Depending on growth conditions the generation times varied from 12 to 140 h. The new Nitrobacter spec. may be one of the most abundant nitrite-oxidizing bacteria in soils, fresh waters and natural as well as artificial stones. For this organism the name Nitrobacter vulgaris is proposed.The type strain is filed with the culture collection of the Institut für Allgemeine Botanik, Universität Hamburg, FRG.     

Ectothiorhodospira halochloris sp. nov., a new extremely halophilic phototrophic bacterium containing bacteriochlorophyll b

A new bacteriochlorophyll b containing phototrophic bacterium was isolated from extremely saline and alkaline soda lakes in Egypt. Enrichment and isolation were performed using a synthetic medium with high contents of sodium carbonate, sodium sulfate and sodium chloride. Photoautotrophic growth occurred with hydrogen sulfide as photosynthetic electron donor. During oxidation of sulfide to sulfate extracellular elemental sulfur globules appeared in the medium. Cells were also capable to grow under photoheterotrophic conditions with acetate, propionate, pyruvate, succinate, fumarate or malate as carbon sources and electron donors. Under these conditions sulfate was assimilated. Optimal growth under the applied experimental conditions occurred at a total salinity of 14–27%, a pH-range between 8.1 and 9.1 and a temperature between 47°C and 50°C. The cells were 0.5–0.6 m wide and, depending on cultural conditions, 2.5–8.0 m long; they were spiral shaped, multiplied by binary fission and were motile by means of bipolar flagella. Intercytoplasmic photosynthetic membranes were present as stacks. Bacteriochlorophyll b was the main photosynthetic pigment; small amounts of carotenoids were mainly present as glucosides of rhodopin and its methoxy derivative. The new organism is described as Ectothiorhodospira halochloris.Dedicated to Professor C. B. van Niel on the occasion of his 80th birthday     

High-frequency somatic embryogenesis from small suspension-cultured clusters of cells of an interspecific hybrid of Oryza

Suspension cultures in which cell clusters were small and had a high capacity for somatic embryogenesis (about 60%) were established from immature panicles of F₁ plants from a cross between Oryza sativa and Oryza latifolia The cells were subcultured at seven-day intervals in a modified N6 medium. The cell clusters were quite small (approximately 30–200m in diameter) after culture for two months in this medium. When small clusters of cells were transferred to N6 medium, that had been diluted with an equal volume of water and supplemented with -naphthalenacetic acid (53 nM), 4-pyridylurea (2.2 M) and sucrose (30 gl^-1), somatic embryogenesis occurred at high frequency (about 60%). The system established in the present work is useful for biochemical and molecular biological research of the somatic embryogenesis in the Gramineae.Abbreviations 2,4-d 2,4-dichlorophenoxyacetic acid - NAA -naphthalenacetic acid - 4-PU 4-pyridylurea - MS Murashige and Skoog (1962)     

Production of thermostable α-galactosidase from thermophilic fungusHumicola sp

The thermophilic fungus,Humicola sp isolated from soil, secreted extracellular -galactosidase in a medium cotaining wheat bran extract and yeast extract. Maximum enzyme production was found in a medium containing 5% wheat bran extract as a carbon source and 0.5% beef extract as a carbon and nitrogen source. Enzyme secretion was strongly inhibited by the presence of Cu²⁺, Ni²⁺ and Hg²⁺ (1mM) in the fermentation medium. Production of enzyme under stationary conditions resulted in 10-fold higher activity than under shaking conditions. The temperature range for production of the enzyme was 37° C to 55°C, with maximum activity (5.54 U ml^–1) at 45°C. Optimum pH and temperature for enzyme activity were 5.0 and 60° C respectively. One hundred per cent of the original activity was retained after heating the enzyme at 60°C for 1 h. At 5mM Hg²⁺ strongly inhibited enzyme activity. TheK _m andV _max forp-nitrophenyl--d-galactopyranoside were 60M and 33.6 mol min^–1 mg^–1, respectively, while for raffinose those values were 10.52 mM and 1.8 mol min^–1 mg^–1, respectively.     

Purification and characterization of hyperthermotolerant leucine aminopeptidase from Geobacillus thermoleovorans 47b

A thermophilic bacterium, which we designated as Geobacillus thermoleovorans 47b was isolated from a hot spring in Beppu, Oita Prefecture, Japan, on the basis of its ability to grow on bitter peptides as a sole carbon and nitrogen source. The cell-free extract from G. thermoleovorans 47b contained leucine aminopeptidase (LAP; EC 3.4.11.10), which was purified 164-fold to homogeneity in seven steps, using ammonium sulfate fractionation followed by the column chromatography using DEAE-Toyopearl, hydroxyapatite, MonoQ and Superdex 200 PC gel filtration, followed again by MonoQ and hydroxyapatite. The enzyme was a single polypeptide with a molecular mass of 42,977.2 Da, as determined by matrix-assisted laser desorption ionization and time-of-flight mass spectrometry, and was found to be thermostable at 90°C for up to 1 h. Its optimal pH and temperature were observed to be 7.6–7.8 and 60°C, respectively, and it had high activity towards the substrates Leu-p-nitroanilide (p-NA)(100%), Arg-p-NA (56.3%) and LeuGlyGly (486%). The K_m and V_max values for Leu-p-NA and LeuGlyGly were 0.658 mM and 25.0 mM and 236.2 mol min^–1 mg^–1 protein and 1,149 mol min^–1 mg^–1 protein, respectively. The turnover rate (k_cat) and catalytic efficiency (k_cat/ K_m) for Leu-p-NA and LeuGlyGly were 10,179 s^–1 and 49,543 s^–1 and 15,470 mM^–1 s^–1 and 1981.7 mM^–1 s^–1, respectively. The enzyme was strongly inhibited by EDTA, 1,10-phenanthroline, dithiothreitol, -mercaptoethanol, iodoacetate and bestatin; and its apoenzyme was found to be reactivated by Co²⁺ .     

Characterization of a pyridine-degrading branched Gram-positive bacterium isolated from the anoxic zone of an oil shale column

Summary From the anoxic zone of an oil shale leachate column three pyridine-degrading bacterial strains were isolated. Two strains were Gram-negative facultative anaerobic rods and one strain was a branched Gram-positive bacterium. The branched Gram-positive strain had the best pyridine-degrading ability. This organism was aerobic, non-motile, catalase positive, oxidase negative, and had no flagellum. The G+C content of the DNA was 66.5 mol%. The major menaquinone was MK-8(H₂). The main cellular fatty acids were saturated and monounsaturated straight chains. This organism contained mycolic acid, meso-diaminopimelic acid, arabinogalactan and glycolyl residues in the cell wall. Due to morphological, physiological and chemotaxonomic characteristics this strain was placed in the genus Rhodococcus. The optimum culture conditions were as follows: temperature 32° C, pH 8.0 and 0.1% v/v of pyridine as sole carbon, energy and nitrogen source. Utilization of pyridine by a batch fermentor culture of Rhodococcus sp. was characterized by a specific growth rate of 0.13 h^–1, growth yield of 0.61 mg cell·mg pyridine^–1 and a doubling time of 5.3 h^–1. Offprint requests to: S.-T. Lee     

Cryopreservation of in vitro-cultured multiple bud clusters of asparagus (Asparagus officinalis L. cv Hiroshimagreen (2n=30) by the techniques of vitrification

Summary A culture line of asparagus forming green bulbous structures consisting of numerous multiple bud clusters designated bud clusters was induced from a meristem culture of asparagus (Asparagus officinalis L.cv. Hiroshimagreen, 2n=30). Small cubic segments (2 mm3) cut from bud clusters were cryopreserved using three different cryogenic protocols. Only vitrification produced very high levels of shoot formation after cooling to –196°C. Segments were treated with a vitrification solution (PVS2) at 25°C for 45 min or at 0°C for 120 min prior to a direct plunge into liquid nitrogen. After rapid warming, the segments were expelled into Murashige and Skoog medium containing 1.2 M sucrose for 10 min and then plated on agar shoot outgrowth medium. The average rate of shoot formation of vitrified segments producing normal shoots was near 90% without any preculture and/or cold-acclimation treatment. Revived segments resumed growth within 3 days and developed about three shoots per segment. In vitro-cultured bud clusters appear promising as material for cryopreserving asparagus germplasm.Abbreviations DMSO dimethyl sulfoxide - PVS 2-vitrification solution - LN liquid nitrogen - IBA 3-indolbutyric acid - BA 6-benzylaminopurine - FDA fluorescein diacetate - DSC differential scanning calorimeter     

Optical resolution of RS-(±)-mandelic acid by Pseudomonas sp.

For the optical resolution of R-(–)-mandelic acid from (±)-mandelic acid, Pseudomonas sp. MA02, which assimilated S-(+)-mandelic acid as carbon and energy source, was isolated from soil. Using the fed-batch culture under optimal condition, R-(–)-mandelic acid was accumulated up to the maximum theoretical yield of 50% (30 g l^–1) and entiomeric excess of 99.4%.     

Thermoanaerobium brockii gen. nov. and sp. nov., a new chemoorganotrophic,caldoactive, anaerobic bacterium

The isolation of a new anaerobic thermophilic bacterium, Thermoanaerobium brockii, from volcanic features is described. Successful enrichment required a complex medium containing glucose or other fermentable sugars and incubation temperatures of 55–80° C. Strains of T. brockii were gram positive, rods of uneven length that existed singly, in pairs, chains or filaments. Electron micrographs of thin sections of cell revealed a monolayered cell wall and a constrictive or pinching off cell division process. The organism was nonsporeforming, obligately anaerobic and chemoorganotrophic. The optimal temperature for growth was 65–70° C, the maxium was below 85° C and the minimum above 35° C. The doubling time at the optimal temperature for growth was about 1 h. The DNA base composition of three strains of T. brockii varied from 30.0–31.4 mol % guanosine plus cytosine. Fermentable carbohydrates included glucose, sucrose, maltose, lactose, cellobiose and insoluble starch. The fermentation products of cells grown on glucose were ethanol, lactic acid, acetic acid, hydrogen and carbon dioxide. Growth of all strains tested was inhibited by fairly low concentrations of cycloserine, penicillin, streptomycin, tetracycline and chloramphenicol. The possible ecological, evolutionary, and industrial significance, and taxonomic relationships of Thermoanaerobium are discussed.Abbreviations TYEG complex medium containing mineral salts, 0.3% yeast extract, 1.0% tryptone and 0.5% glucose - O.D. optical density - G+C guanosine plus cytosine     

Optimum growth conditions and light utilization efficiency of Spirulina platensis (= Arthrospira fusiformis) (Cyanophyta) from Lake Chitu, Ethiopia

Spirulina platensis (= Arthrospira fusiformis) was isolated from Lake Chitu, a saline, alkaline lake in Ethiopia, where it forms an almost unialgal population. Optimum growth conditions were studied in a turbidostat. Cultures grown in modified Zarrouk's medium and exposed to a range of light intensities (20–500 µmol photons m^–2s^–1) showed a maximum specific growth rate (µ_max) of 1.78 d^–1. Quantum yield for growth (µ) was 3.8% at the optimum light for growth of 330 µmol photons m^–2s^–1, and ranged from 2.8 to 9.4%. With increase in irradiance, the chlorophyll a concentration decreased, and the carotenoids/chlorophyll a ratio increased by a factor of 2.4. The phosphorus to carbon ratio (P/C) showed some variation, while the nitrogen to carbon ratio (N/C) remained relatively constant, thus causing fluctuations in the N:P ratio (7–11) of cells. An optimum N:P ratio of about 7 was attained in cells growing at the optimum light for growth. Results from the continuous culture experiments agreed well with maximum values of photosynthetic efficiency given in the literature for natural populations of S. platensis in the soda lakes of East Africa, Lake Arenguade (Ethiopia), and Lake Simbi (Kenya).     

Plant regeneration from cultured protoplasts of the cooking banana cv. Bluggoe (Musa spp., ABB group)

Summary Suspensions of embryogenic cells of a triploid banana (Musa spp., cv. Bluggoe) were initiated from the uppermost part of meristematic buds, and used as protoplast source. After 20 weeks in culture, the suspension contained a mixture of globular structures or globules and embryogenic cell clusters, as well as single cells. Two types of protoplasts were obtained from embryogenic suspension culture: small (20–30 m) and larger (30–50 m) protoplasts with a dense cytoplasm and large starch grains respectively. The small protoplasts probably originated from embryogenic cell clusters, and also from pseudocambial cells of globules, while larger protoplasts were probably released from oval starchy cells and those of the globule peripheral area. In co-culture with a suitable feeder, consisting of suspensions of diploid banana cells, the protoplasts of triploid banana reformed the cell wall within 24 h and underwent sustained divisions leading to the formation of small clusters of 2–3 cells within 7 days. The latter developed directly into embryos without passing through an apparent callus phase. 10% of such embryos gave rise to plantlets when subcultured in 2.2 M 6-benzylaminopurine and 2 M 4 amino-3,5,6-trichloropicolinic acid for 1 week, before transfer to MS medium containing 10 M 6-benzylaminopurine. The rest of the embryos underwent intensive direct secondary embryogenesis which could lead to the formation of plantlets with a frequency of up to 50% upon further transfer to hormone-free medium.Abbreviations BAP 6-benzylaminopurine - MS Murashige and Skoog (1962) medium - 2,4-D dichlorophenoxyacetic acid - UV ultraviolet light - FDA fluorescein diacetate - MES 2-(N-morpholino)ethanesulfonic acid - Picloram 4 amino-3,5,6-trichloropicolinic acid     

Nutritional control of copper uptake in the cyanobacteriumNostoc calcicola Bréb

Summary The kinetics of Cu uptake in nutritionally starved cells of the diazotrophic cyanobacteriumNostoc calcicola Bréb. have been compared with those in cells recovering from starvation. Unstarved cyanobacterial cells assimilated 97.0 nmol Cu mg^–1 protein within 1 h when incubated in medium containing 40 M Cu. Uptake was markedly inhibited in carbon-starved cells and, to a lesser extent, in cells starved of nitrogen or sulphur. The intracellular concentrations of protein and photopigments were markedly lower in cells starved of carbon, nitrogen, sulphur or phosphorus, whilst that of carbohydrate was lower in cells starved of carbon, sulphur or phosphorus, but almost doubled in cells starved of nitrogen. The ability to assimilate Cu was partially restored in cells after 72 h of recovery from phosphorus or sulphur deprivation, but showed little improvement during recovery from carbon or nitrogen starvation. A possible role of phosphorus in regulating Cu transport and accumulation is discussed.     

<Emphasis Type="Italic">Halobacillus locisalis</Emphasis> sp. nov., a halophilic bacterium isolated from a marine solar saltern of the Yellow Sea in Korea

A Gram-positive, motile, endospore-forming and rod-shaped halophilic bacterial strain MSS-155 (KCTC 3788 and KCCM 41687) was isolated from a marine solar saltern of the Yellow Sea in Korea and was subjected to a polyphasic taxonomic study. This organism grew at temperature of 10.0–42.0°C with an optimum of 35°C. Strain MSS-155 grew optimally in the presence of 10% NaCl and did not grow in the absence of NaCl. The cell wall peptidoglycan type of strain MSS-155 was A4 based on l-Orn-d-Asp. Strain MSS-155 was also characterized chemotaxonomically by having menaquinone-7 (MK-7) as the predominant isoprenoid quinone and anteiso-C_15:0 as the major fatty acid. The DNA G+C content was 44.0 mol%. Phylogenetic analysis based on 16S rDNA sequences showed that strain MSS-155 falls within the radiation of the cluster comprising Halobacillus species. Levels of 16S rDNA sequence similarity between strain MSS-155 and the type strains of four Halobacillus species were in the range 97.6–98.8%. Strain MSS-155 exhibited levels of DNA-DNA relatedness of 6.2–11.2% to the type strains of Halobacillus species described previously. On the basis of phenotypic properties, phylogeny, and genomic data, strain MSS-155 should be placed in the genus Halobacillus as a member of a novel species, for which we propose the name Halobacillus locisalis sp. nov.Communicated by W.D. Grant     

Physiology of a Methanobacterium strain AZ

A methanogenic bacterium using H₂ and CO₂ as sole energy and carbon source has been isolated in pure culture from digested sludge. Its colonies on mineral agar are translucent, convex, circular with entire margins and yellow to brownish in colour. Cells are gram-positive, non motile and appear as straight cods, normally about 3 m long. A marked pleomorphism depending on the media was observed. The organism is chemolithoheterotrophic, has a pH optimum of 7.0 and an optimal temperature for growth of 33–40°C; no growth occurs above 45°C. The generation time at optimal conditions is less than 5 h. Cysteine must be supplied in the growth medium. It can act as sole sulfur source. The addition of sulfide accelerates the growth at an optimum concentration of 10^-4 to 10^-5 molar. A growth factor, not identical with SH-coenzyme M, occurring in anaerobic sewage sludge and yeast extract shows a stimulatory effect. 7.0–8.2% of the total carbon dioxide uptake is assimilated and 11.2% of the energy obtained from the reduction of carbon dioxide to methane is refound in the caloric value of the biomass. 0.01 ppm of dissolved oxygen completely inhibits growth and methane production. However, the bacteria do not loose their viability when exposed to high oxygen concentrations. Further informations are needed before this organism (DSM 744) is specifically identified.List of Abbreviations TOC total organic carbon - DOC dissolved organic carbon - POC particulate organic carbon     

Factors affecting in vitro microrhizome production in turmeric

In vitro microrhizome production was obtained in turmeric (Curcuma longa Linn.). Freshly sprouted buds with small rhizome portions excised from stored mature rhizomes were cultured on semi-solid culture initiation medium –- MS basal medium + 0.88 M BAP (6-benzylaminopurine) + 0.92 M kinetin + 5% coconut water + 2% sucrose + 0.5% agar –- resulting in bud elongation. Multiple shoots were produced from these elongated buds by culturing in liquid shoot multiplication medium –- MS basal medium + 2.2 M BAP + 0.92 M kinetin + 5% coconut water + 2% sucrose –- at 25±1°C and 16-h light (at 11.7 mol m^–2 s^–1)/8-h dark cycles. Clumps of four to five multiple shoots/single shoots were used in various experiments. Cultures were incubated in the dark at 25±1°C. Half strength MS basal medium supplemented with 80 g l^–1 sucrose was found to be optimal for microrhizome production. Cytokinin BAP had an inhibitory effect on microrhizome production. At the highest concentration of BAP tried (35.2 M) microrhizome production was totally inhibited. Microrhizome production depended on the size of the multiple shoots used. Microrhizomes produced were of a wide range in size (0.1–2.0 g) and, readily regenerated when isolated and cultured in vitro on culture initiation medium or shoot multiplication medium. Under in vivo conditions, small (0.1–0.4 g), medium (0.41–0.8 g) and big (>0.81 g) microrhizomes regenerated. Plantlets developed from big microrhizomes grew faster.     

Energy conservation during aerobic growth in Paracoccus denitrificans

Paracoccus denitrificans was aerobically grown in chemostat culture with succinate or gluconate as carbon source. Due to the presence of two phosphorylation sites in the respiratory chain and the absence of branching, theoretical P/O ratios of 1.71 and 1.82 were calculated for cells growing respectively with succinate and gluconate as carbon source. Using these data, 95% confidence intervals for the P/O ratio were determined, via a mathematical model, at 0.91–1.15 and 1.00–1.37 for sulphate-limited cultures, with respectively succinate and gluconate as carbon source.These results and measurements of P/O ratios in membrane particles and of proton translocation in whole cells have led to the conclusion that site I phosphorylation is affected under sulphate-limited conditions.Under conditions of carbon source-limitation the endogenous H⁺/O ratio is about 7–8. Average values of 3.40 and 4.78 were respectively found for sulphate-limited succinate- and gluconate grown cells. For starved cells, oxidizing succinate as exogenous substrate, the H⁺/O ratios were determined at about 3–4, independent of the growth limiting factor. It is concluded that the number of protons ejected per pair of electrons per energy-conserving site (H⁺/site ratio) is about 3–4, instead of 2 as postulated by the chemiosmotic hypothesis.     

Acetonema longum gen.nov.sp.nov., an H2/CO2 acetogenic bacterium from the termite,Pterotermes occidentis

A previously undescribed, H₂-oxidizing CO₂-reducing acetogenic bacterium was isolated from gut contents of the wood-feeding termite, Pterotermes occidentis. Cells of representative strain APO-1 were strictly anaerobic, Gram-negative, endospore-forming motile rods which measured 0.30–0.40×6–60 m. Cells were catalase positive, oxidase negative, and had 51.5 mol percent G+C in their DNA. Optimum conditions for growth on H₂+CO₂ were at 30–33°C and pH (initial) 7.8, and under these conditions cells formed acetate according to the equation: 4 H₂+2 CO₂CH₃COOH+2 H₂O. Other energy sources supporting good growth of strain APO-1 included glucose, ribose, and various organic acids. Acetate and butyrate were major fermentation products from most organic compounds tested, however propionate, succinate, and 1,2-propanediol were also formed from some substrates. Based on comparative analysis of 16S rRNA nucleotide sequences, strain APO-1 was related to, but distinct from, members of the genus Sporomusa. Moreover, physiological and morphological differences between strain APO-1 and the six known species of Sporomusa were significant. Consequently, it is proposed herewith that a new genus, Acetonema, be established with strain APO-1 as the type strain of the new species, Acetonema longum. A. longum may contribute to the nutrition of P. occidentis by forming acetate, propionate and butyrate, compounds which are important carbon and energy sources for termites.     

DNA base composition heterogeneity in some agarophytes (Gracilariales,Rhodophyta) from Mexico and the Philippines

Estimates of nuclear DNA base composition by determination of thermal denaturation temperatures (T_m) indicate guanine + cytosine (G + C) levels of 35.4–46.8% for ten species of the Gracilariaceae, representing the generaGracilaria andHydropuntia. T_m values were found to be reproducible with variation among most samples and replicates of less than 1 °C and 2 mol%. Interspecific variation in G + C values was less than 11.4% amongGracilaria species. Calculation of intragenomic base pair composition distribution based on mid-resolution thermal denaturation (A 1 °C/min with 4s interval H and dT logging) indicated an inverse relationship between maximum similarity values and taxonomic rank. Intraspecific (population level) maximum similarity (homology) values were estimated to range from 79–90% inGracilaria tikvahiae (4 isolates). Interspecific values of 46–69% were found in 13 species ofGracilaria. Nucleotide distribution similarity values for the Gracilariaceae are compared with previous information for genome organization and complexity, genome size and karyotype patterns.Author for correspondence