Sequence analysis of duplicated actin genes in Lagenidium giganteum and Pythium irregulare (Oomycota)

Southern analysis of genomic DNA identified multiple-copy actin gene families in Lagenidium giganteum and Pythium irregulare (Oomycota). Polymerase chain reaction (PCR) protocols were used to amplify members of these actin gene families. Sequence analysis of genomic coding regions demonstrated five unique actin sequences in L. giganteum (Lg-Ac 1, 2, 3, 4, 5) and four unique actin sequences in P. irregulare (Pi-Acl, 2, 3, 4); none were interrupted by introns. Maximum parsimony analysis of the coding regions demonstrated a close phylogenetic relationship between oomycetes and the chromophyte alga Costaria costata. Three types of actin coding regions were identified in the chromophyte/oomycete lineage. The type 1 actin is the single-copy coding region found in C. costata. The type 2 and type 3 actins are found in the oomycetes and are the result of a gene duplication which occurred soon after the divergence of the oomycetes from the chromophyte algae. The type 2 coding regions are the single-copy sequence of Phytophthora megasperma, the Phytophthora infestans actB gene, Lg-Ac5 and Pi-Ac2. The type 3 coding regions are the single-copy sequence of Achlya bisexualis, the P. infestans actA gene, Lg-Ac1, 2, 3, 4 and Pi-Acl, 3, 4. Correspondence to: D. Bhattacharya     

Reduction of Cr(VI) by Desulfovibrio vulgaris and Microbacterium sp.

Many industrial wastes contain Cr(VI), a carcinogen and mutagen, the toxicity of which can be ameliorated by reduction to Cr(III). Microbacterium sp. NCIMB 13776 andDesulfovibrio vulgaris NCIMB 8303 reduced Cr(VI) to Cr(III) anoxically using 25 mM sodium citrate buffer (pH 7), with 25 mM sodium acetate and 25 mM sodium formate as electron donors at 30 °C, under which conditions the rates of reduction of 500 M sodium chromate were 77 and 6 nmol h^–1 mg dry cell wt for D. vulgaris and Microbacterium sp., respectively, these being increased to 127 and 17 nmol h^–1 mg dry cell wt in the presence of 20 mM MOPS/NaOH buffer.     

Biological control of alligatorweed (Alternanthera philoxeroides) with a Fusarium sp.

Alligatorweed (Alternanthera philoxeroidesG.) has become a serious weed in different crops in China. A fungal pathogen was found in Chongqing and Sichuan Provinces and was identified as a species in the Fusarium genus. The fungus produced macroconidia and chlamydospores abundantly on potato sucrose agar (PSA) plates. The bestconidial production and germination and colonygrowth of Fusarium sp. were at 23–31°C and pH 6.7–7.0. Light period and flooding did not affect fungal growth and conidium formation. The herbicides, glyphosate and paraquat, inhibited the fungal development in vitro. The fungus did not affect seed germination and seedling growth of paddy rice, wheat, maize, oilseed rape and broad bean inlaboratory or greenhouse trials. Inoculum density and wetness duration influenced the efficiency of Fusarium sp. to control alligatorweed; a concentration of 1.0 × 10⁵ spores^–1 ml and 12 h of high humidity duration after inoculation produced goodinfections on the weed at 23°C in the laboratory. When the fungus was applied to alligatorweed grown in greenhouse and in the field, good biocontrol efficiency was obtained: the plants started to wilt after four to five (greenhouse) or six days (field), and were killed 9–10 (greenhouse) or 13–14 (field) days after spraying the fungal inoculum. This was similar to the control efficiency resulting from glyphosate treatment. Therefore, this Fusarium sp. appeared to be a good candidatefor further studies and a promising biocontrol agent to manage alligatorweed in some terrestrial and aquatic crops.     

Evidence for a peptidoglycan envelope in the cyanelles of Glaucocystis nostochinearum Itzigsohn

Glaucocystis nostochinearum is a eukaryotic organism with chloroplasts that have usually been assumed to be cyanelles — i.e., endosymbiotic cyanobacteria. Previous attempts by others to support this assumption by demonstrating the presence of a limiting peptidoglycan envelope have been unsuccessful.In the present study disruption of intact Glaucocystis cells with a glass tissue homogenizer permitted the isolation of the uniquely-shaped cyanelles. That these cyanelles were lunited by a peptidoglycan-containing envelope was concluted from the following evidence: (1) stability of isolated cyanelles in distilled water as determined by the preservation of their intactness and peculiar asymmetrical shape; (2) lysozyme sensitivity as demonstrated by lysis of isolated cyanelles when treated with low concentrations of lysozyme; (3) inhibition of the lysozyme-mediated lysis by N-acetyl-glucosamine-2, a known competitive inhibitor of lysozyme, (4) visualization of a thin, electron dense layer between the two limiting membranes around the cyanelle, and (5) isolation and identification of the peptidoglycan-specific amino acid, diaminopimelic acid, from the cyanelles.     

Four novel yeasts from decaying organic matter: Blastobotrys robertii sp. nov., Candida cretensis sp. nov., Candida scorzettiae sp. nov. and Candida vadensis sp. nov

Four novel yeast species are described, two from decaying mushrooms, viz. Candida cretensis and Candida vadensis, and two from rotten wood, viz. Blastobotrys robertii and Candida scorzettiae. Accession numbers for the CBS and ARS Culture Collections, and GenBank accession numbers for the D1/D2 domains of the large subunit of ribosomal DNA are: B. robertii CBS 10106^T, NRRL Y-27775, DQ839395; C. cretensis CBS 9453^T, NRRL Y-27777, AY4998861 and DQ839393; C. scorzettiae CBS 10107^T, NRRL Y-27665, DQ839394; C. vadensis CBS 9454^T, NRRL Y-27778, AY498863 and DQ839396. The GenBank accession number for the ITS region of C. cretensis is AY498862 and that for C. vadensis is AY498864. C. cretensis was the only species of the four that displayed fermentative activity. All four type strains grew on n-hexadecane. C. scorzettiae is the only one of the new species that assimilates some phenolic compounds, viz. 3-hydroxy derivatives of benzoic, phenylacetic and cinnamic acids, but not the corresponding 4-hydroxy acids. This is indicative of an operative gentisate pathway.     

Degradation of carbazole and its derivatives by a Pseudomonas sp.

Carbazole, carbazoles with monomethyl or dimethyls substituted on different positions (C₁-carbazoles or C₂-carbazoles), and benzocarbazoles, as toxic and mutagenic components of petroleum and creosote contamination, were biodegradable by an isolated bacterial strain Pseudomonas sp. XLDN4-9. C₁-carbazoles were degraded in preference to carbazole and C₂-carbazoles. The biodegradation of C₁-carbazoles or C₂-carbazoles was influenced by the positions of methyl substitutions. Among C₁-carbazole isomers, 1-methyl carbazole was the most susceptible. C₂-carbazole isomers with substitutions on the same benzo-nucleus were more susceptible at a concentration of less than 3.4 μg g⁻¹ petroleum, especially when harboring one substitution on position 1. In particular, 1,5-dimethyl carbazole was the most recalcitrant dimethyl isomer.     

Reduction of Cr(VI) by a Bacillus sp.

A Bacillus sp. RE was resistant to chromium and reduced Cr(VI) without accumulating chromium inside the cell. When Cr(VI) was 10 and 40 μg ml⁻¹, >95% of the total Cr(VI) was reduced in 24 and 72 h of growth, respectively, whereas at 80 μg Cr(VI) ml⁻¹ only 50% of Cr(VI) was reduced. However growth was not affected; the cell mass was 0.7–0.8 mg ml⁻¹ in all cases. The cell-free extract showed Cr(VI) reducing enzyme activity which was enhanced (>5 fold) by NADH and NADPH. Like whole cells the enzyme also reduced Cr(VI) with decreasing efficiency on increasing Cr(VI) concentration. The enzyme activity was optimal at pH 6.0 and 30 °C. The enzyme was stable up to 30 °C and from pH 5.5 to 8, but from pH 4 to 5 the enzyme was severely destabilized. Its K_m and V_max were 14 μm and 3.8 nmol min⁻¹ mg⁻¹ respectively. The enzyme activity was enhanced by Cu²⁺ and Ni²⁺ and inhibited by Hg²⁺. Received 21 September 2005; Revisions requested 5 October 2005; Revisions received 16 November 2005; Accepted 16 November 2005     

Candida alocasiicola sp. nov., Candida hainanensis sp. nov., Candida heveicola sp. nov. and Candida musiphila sp. nov., novel anamorphic, ascomycetous yeast species isolated from plants

In a taxonomic study on the ascomycetous yeasts isolated from plant materials collected in tropical forests in Yunnan and Hainan Provinces, southern China, four strains isolated from tree sap (YJ2E(T)) and flowers (YF9E(T), YWZH3C(T) and YYF2A(T)) were revealed to represent four undescribed yeast species. Molecular phylogenetic analysis based on the large subunit (26S) rRNA gene D1/D2 domain sequences showed that strain YJ2E(T) was located in a clade together with Candida haemulonii and C. pseudohaemulonii. Strain YF9E(T) was most closely related to C. azyma and strain YWZH3C(T) to C. sorbophila and C. spandovensis. Strain YYF2A(T) was clustered in a clade containing small-spored Metschnikowia species and related anamorphic Candida species. The new strains differed from their closely related described species by more than 10% mismatches in the D1/D2 domain. No sexual states were observed for the four strains on various sporulation media. The new species are therefore assigned to the genus Candida and described as Candida alocasiicola sp. nov. (type strain, YF9E(T) = AS 2.3484(T) = CBS 10702(T)), Candida hainanensis sp. nov. (type strain, YYF2A(T) = AS 2.3478(T) = CBS 10696(T)), Candida heveicola sp. nov. (type strain, YJ2E(T) = AS 2.3483(T) = CBS 10701(T)) and Candida musiphila sp. nov. (type strain, YWZH3C(T) = AS 2.3479(T) = CBS 10697(T)).     

Phyllodiaptomus praedictus n. sp. (Copepoda,Calanoida) from Thailand

Phyllodiaptomus praedictus n. sp. is described from the Bangkok area. It might be endemic to Thailand. It belongs to the blanci-group of species, and is most closely related to P. annae and P. wellekensae.     

Microbial Metabolism of Quinoline by Comamonas sp.

An aerobic bacterial strain which can use quinoline as the sole carbon and energy source has been isolated from activated sludge and identified as Comamonas sp. The microbial metabolism of quinoline by this strain has been investigated. A pH 8 and a temperature of 30 °C were the optimum degradation conditions of quinoline. Five intermediates including 2-oxo-1,2-dihydroquinoline, 5-hydroxy-6-(2-carboxyethenyl)-1H-2-pyridone, 6-hydroxy-2-oxo-1,2-dihydroquinoline, 5,6-dihydroxy-2-oxo-1,2-dihydroquinoline, and 8-hydroxy-2-oxo-1,2-dihydroquinoline were found during quinoline biodegradation. The presence of these intermediates suggested that at least two pathways were involved for quinoline degradation by Comamonas sp. and a reasonable degradation route was proposed to account for the intermediates observed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Eodiaptomus shihi n. sp. (Copepoda,Calanoida) from central India

Eodiaptomus shihi n. sp. is described from the material collected from Gandhisagar reservoir, River Narmada and a hillpool in the Madhya Pradesh State of central India. The salient features of this new species include: in female, the left metasomal wing is strongly developed, and the endopodite in leg 5 is 2-segmented and longer than first exopodite-segment; in male, the right caudal ramus is armed with a tooth-like chitinous structure near ventro-distal region, and in right leg 5, the basis is devoid of any hyaline lobe on inner margin and the first exopodite-segment is produced into a large spinous process at disto-outer corner.     

Atkinsiella infection in the rotiferBrachionus plicatilis

Atkinsiella infection in the rotiferBrachionus plicatilis has not hitherto been reported. The first known case of this infection is described herein. The fungal growth in vitro was inhibited by 0.05µg/ml of malachite green or 15.6 ppm of formalin. It was experimentally demonstrated that the fungus is pathogenic to the swimming crab,Portunus trituberculatus in the zoeal stage.     

Secondary carotenoids formation by the green alga Chlorococcum sp.

The effects of temperature, pH and oxygen level onsecondary carotenoids (SC) accumulation in Chlorococcum sp. were investigated. The optimaltemperature, pH and oxygen level for the yields ofsecondary carotenoids and astaxanthin were35 ^°C, pH 8 and 10% (v/v), respectively. Whilethe ratio (R) of products containing hydroxyl group(s) to those containing keto group (s) increased withthe increase of temperature and pH, R value decreasedwith the level of oxygen. These results indicate thathigher temperature and pH favor the introduction ofhydroxyl group to the corresponding substrates,however, under high oxygen level keto group wasrelatively easy to be added compared to the additionof hydroxyl.     

Simultaneous production of hydrogen and bioflocculant by Enterobacter sp. BY-29

Hydrogen and a bioflocculant could be produced simultaneously by anaerobic culture of Enterobacter sp. BY-29. For production of hydrogen and the bioflocculant by cell culture of the bacterium in batch cultures, cultivation at 37 °C in a medium containing glucose as a carbon source and Polypepton as a nitrogen source was found to be suitable. In continuous production of hydrogen and the bioflocculant by cell culture or immobilized cells of the bacterium, the hydrogen production rate and hydrogen yield by the immobilized cells on porous glass beads in stirred and column reactors were higher than those by the cell culture in a stirred reactor. However, production of the bioflocculant by the cell culture was superior to that by the immobilized cells in continuous production.     

Light-path length and population density in photoacclimation of Nannochloropsis sp. (Eustigmatophyceae)

Photoacclimation in the marine eustigmatophyte Nannochlropsis sp., used extensively as a food chaincomponent in aquaculture, was studied both in thelaboratory and outdoors. Cell-chlorophyll andcarotenoids were used as markers to assessphotoacclimation to strong light, as well as todecreasing growth irradiance due to cellproliferation. Focusing on practical aspects involvedin mass cultivation, three different approaches wereused as follows: (a) cultures initially exposed to lowlight (150 mol photon m^-2 s^-1) thentransferred to strong light (1000 to 3000 molphoton m^-2 s^-1); (b) initially low celldensity cultures grown in reactors of differentlight-paths, exposed to strong PFD, in the laboratoryand outdoors; (c) initially low or high cell densitycultures exposed to strong light. As has already beenestablished in many reports, cell-chlorophyllrepresented a sensitive parameter in assessing cellresponse to changes in the intensity of the lightsource as well as to modifications in the light regimeto which the cells were exposed. Cell-chlorophyllconcentration sharply decreased initially upontransferring the culture from low PFD cell^-1 tohigh PFD cell^-1 due to either culture dilution(i.e. decrease in cell density and mutual shading) orto an increase in PFD. After some 7 days ofphotoacclimating to 2000 and 3000 mol photonm^-2 s^-1, chlorophyll a content began to riseat a much faster rate than cell number, which alsoincreased in response to the higher irradiance.Cell-chlorophyll in the culture exposed to 2000mol photon m^-2 s^-1 increased afteracclimation earlier and at a faster rate than in theculture exposed to 3000 mol photon m^-2s^-1, indicating the later irradiance affected astronger stress. The length of the reactor's lightpath exerted a decisive effect on cell response tostrong light through its influence on the light regimein the culture. Upon a sharp increase in PFD,carotenoids in the 1-cm reactor increased in muchhigher rate than chlorophyll, compared with the 3-cmlight path reactors. This marked difference in cellresponse to a shift-up in light was attributed to thevast variations in the light regime associated withdifferences in the length of the light path and areal density. Growth oflow cell density cultures ceased temporarily upontransfer to strong light, in contrast with high celldensity cultures transferred to strong light, whichcontinued growth without a lag.     

Arctodiaptomus (Rhabdodiaptomus) michaeli n.sp. (Copepoda,Calanoida) from Kashmir,India

Arctodiaptomus (Rhabdodiaptomus) michaeli n.sp. is described from the material collected in Alapathar Lake and Marsar Lake of Kashmir, India.     

Production and characteristics of a bioflocculant produced by Bacillus sp. F19

A bioflocculant-producing bacterium isolated from soil was identified as Bacillus sp. and the bioflocculant produced was named MBFF19. Effects of physico-chemical conditions including pH, carbon sources and nitrogen sources on MBFF19 production were studied. Chemical analyses of the purified bioflocculant MBFF19 indicated that it was a sugar-protein derivative, composed of neutral sugar (3.6%, w/w), uronic acid (37.0%, w/w), amino sugars (0.5%, w/w) and protein (16.4%, w/w). The two neutral sugar components were mannose and glucose and the molar ratio was 1.2:1. Infrared spectrophotometry analysis revealed that MBFF19 contained carboxyl, hydroxyl and methoxyl groups in its structural. Flocculating properties of bioflocculant MBFF19 was examined using kaolin, activated carbon and fly coal suspension. Cation supplement had no positive effects on the flocculating activity whereas the presence of Fe3+ inhibited flocculation. Influences of pH and bioflocculant dosage on the flocculation were also examined.     

An immunofluorescence study of mitosis in a mite-pathogen,Neozygites sp. (Zygomycetes: Entomophthorales)

Summary Mitosis in a mite-pathogenic species ofNeozygites (Zygomycetes: Entomophthorales) was investigated by indirect immunofluorescence microscopy using an antibody against -tubulin for visualization of microtubules (MTs). DAPI and rhodamine-conjugated phalloidin were used to stain chromatin and actin, respectively. Salient features of mitosis inNeozygites sp. are (1) a strong tendency for mitotic synchrony in any given cell, (2) conical protrusions at the poles of metaphase and anaphase nuclei revealed by actin staining, (3) absence of astral and other cytoplasmic MTs, (4) a spindle that occupies most of the nuclear volume at metaphase, (5) a spindle that remains symmetrical throughout most of mitosis, (6) kinetochore MTs that shorten during anaphase A, (7) a central spindle that elongates during anaphase B, pushing the daughter nuclei into the cell apices, and (8) interpolar MTs that continue to elongate even after separation of the daughter nuclei. Cortical cytoplasmic MTs are present in a few interphasic and post-cytokinetic cells. The data presented show thatNeozygites possesses features unique to this genus and support the erection of theNeozygitaceae as a separate family in theEntomophthorales.Abbreviations DAPI 4,6-diamidino-2-phenylindole - MT microtubule - SPB spindle pole body