<Emphasis Type="Italic">In vitro</Emphasis> screening of sugarcane to evaluate smut susceptibility

Tissue-cultured plantlets of three sugarcane (Saccharum spp.) cultivars having a known field smut reaction were screened for susceptibility to Ustilago scitaminea H&P Sydow. Plantlets were inoculated with 0.5 l of a suspension of equally mixed quantities of plus and minus mating type sporidia of U. scitaminea at concentrations ranging from 1×10¹ to 1×10⁶ cells. Fungal sori (whips) were produced in cultivar N12 (intermediate) 6weeks following inoculation with 1×10⁵ mixed sporidia and thereafter in cultivar NCo310 (susceptible) but not in cultivar N19 (resistant). Sori bearing teliospores were produced up to 3months following inoculation and incubation at 26°C. No sori were produced at mixed sporidial concentrations lower than 1×10⁵cells. The in vitro soral production in cultivars N19, N12 and NCo310 was 0, 27.5 and 47.5% respectively. Plantlets inoculated with 1×105sporidia of only one mating-type did not produce sori in any of the three cultivars tested. Blind scoring of an unknown sugarcane cultivar by this method corresponded exactly with its field smut rating.     

Relation between trace element levels in plasma and myocardium during coxsackievirus B3 myocarditis in the mouse

During most infections the plasma levels of trace elements change, but it is not clear if this reflects changes in the infected tissues. Coxsackievirus B3 (CB3) infection may result in viral replication, subsequent inflammation and changed trace element levels in the myocardium. In the present study, the trace element levels in the plasma and heart of adult male A/J mice were determined during the pre-inflammatory stage (day 4) of CB3 myocarditis for the following trace elements: aluminium (Al), arsenic (As), calcium (Ca), cobalt (Co), copper (Cu), iron (Fe), magnesium (Mg), manganese (Mn), selenium (Se), silver (Ag), vanadium (V) and zinc (Zn). The severity of the infection was assessed through clinical signs of disease and trace element levels were measured through inductively-coupled plasma mass-spectrometry (ICP-MS). In the heart, the levels decreased for V (59%; p<0.01), Co (38%; p<0.01), Al (81%; p< 0.01), As (66%; p<0.01) and Se (16%; p<0.01). Increased levels were detected for Mn (13%; p<0.05), Fe (48%; p<0.01), Cu (34%; p<0.01) and Ag (46%; p< 0.01). In the plasma, decreases were detected in the level of Zn (32%; p<0.05), whereas increases were seen in Mn (362%; p<0.05), Fe (272%; p<0.01), Co (71%; p<0.05), Cu (25%; n.s.) and Mg (43%; p<0.01) levels. A correlation was found between the levels in plasma and myocardium for Co (r _s=–0.636; p<0.05), Fe (r _s=0.764; p<0.05), Mn (r _s=0.682; p<0.05) and Mg (r _s=–0.791; p<0.05). Thus, determination of some of these trace elements in the plasma may be useful to indicate target tissue involvement in the early pre- inflammatory stage of an infectious disease. Some of these elements are important nutrients for the immune system, while others may be associated with the development of disease complications, such as cardiac arrhythmias.     

Proliferation and germination of somatic embryos from embryogenic suspension cultures in <Emphasis Type="Italic">Phœnix dactylifera</Emphasis>

The present study demonstrates a procedure for the rapid development of a high number of somatic embryos from embryogenic suspension culture. This method might be efficient for mass propagation of Phnix dactylifera L. Embryogenic callus placed in liquid medium with 10^–5M ABA yielded an average 72 embryos per 100ml of culture medium within 2months, while those placed on solid medium yielded an average of 33, 20 and 16 embryos per 100ml of culture medium respectively for 10^–7, 10^–6 and 10^–5 M ABA after 4months. The combination of 2,4-DIchlorophenoxyacetic acid (2,4-D) (4.5×10^–7M), glutamine (6.7×10^–4M), and ABA (10^–5M) (L8 liquid medium) showed a beneficial effect on somatic embryos production compared to 2,4-D and glutamine alone, while this combination significantly (p<0.05) increased the accumulation of storage proteins (144 and 138mgg^–1 DW respectively for Jihel and Bousthami noir cultivars) in somatic embryos. The somatic embryos which underwent maturation on medium containing only 4.5×10^–7M 2,4-D and 10^–5M ABA (L6 liquid medium) accumulated more sugars (292 and 265mgg^–1 DW respectively for Jihel and Bousthami noir) than those matured on any other liquid medium. Histological studies revealed that somatic embryos (developed in L6 and L8 liquid media) accumulated less reserve compounds (proteins and sugars) than zygotic embryos. The addition of activated charcoal (0.25 and 0.5gl^–1) and phytagel^® (2.5gl^–1) to the germination medium may be useful for enhancing the germination of Phnix dactyliferasomatic embryos.     

Flexibility on the karyotype evolution in bitterlings (Pisces, Cyprinidae)

In bitterlings (Acheilognathinae) C- and Ag-banding karyotypes of 6 species-subspecies collected in China and South Korea were analyzed. The chromosomal constitution of 2n=46 (4SM+42ST) in Rhodeus atremius fangi was quite different from that of 2n=48 (8M+20SM+20ST) in other species-subspecies in Rhodeus. It was concluded from the analysis of banded chromosomes that the increase in number of ST during the karyotype change from 2n=48 to 2n=46 was achieved by a series of pericentric inversions from 24 M-SM to 24 ST, and the decrease in the diploid number was caused by an additional tandem fusion of 4 ST chromosomes, forming a new ST pair in the 2n=46 karyotype. The karyotype of Tanakia koreensis, T. signifer, and Acheilognathus macropterus is 2n=48 (8M+20SM+20ST), 2n=48 (8M+20SM+14–16ST+4–6 A), 2n=44 (14M+16SM+14ST), respectively. In R. ocellatus ocellatus, T. koreensis, T. signifer and A. macropterus, karyotype changes from 2n=48 to 2n=44 due to centric fusion and inversion have also been estimated. It was suggested that C-banding heterochromatin was greatly concerned with the karyotype evolution in bitterlings.     

Light utilization and competition between Echinacea purpurea, Panicum virgatum and Ratibida pinnata under greenhouse and field conditions

Competitive interactions were compared under field and greenhouse conditions for three representative tallgrass prairie species, Echinacea purpurea, Panicum virgatum and Ratibida pinnata. These were planted in monoculture and in mixtures of two species using a replacement series design with groups of four or eight plants. Competition was determined from shoot dry weight data collected during 120days in the greenhouse and after 415days in the field. Yields declined with increased density of a single species in the greenhouse from 40days onward and in the field. Relative yields were up to 100% higher in mixtures than in monocultures for all species early on in the greenhouse experiment. Later in the experiment and in the field relative yields of E.purpurea decreased in the presence of P.virgatum and R.pinnata, whereas relative yields of these two species increased in the presence of E.purpurea. There were correlations in relative yield between the field and the greenhouse experiment at 80 and 120days. In the greenhouse P.virgatum maintained higher net assimilation rates than the other species. Relative growth rates of all species were higher in monoculture and in mixtures up to 40days, after which they declined, especially for E.purpurea in mixtures. In the field, higher light intensities occurred in pure stands of E.purpurea than in mixed stands with other species. The order of competitive ability that was apparent from these field and greenhouse studies, P.virgatum=R.pinnata>E.purpurea, could be partially explained by photosynthetic rates in relation to canopy light interception.     

Recombinant Components of the EcoRII Restriction–Modification System: Restriction Endonuclease Can Interact with DNA · RNA Duplexes

To obtain recombinant restriction endonuclease (R) and methylase (M) of the EcoRII restriction–modification system, bacterial strains overproducing their functional hexahistidine derivatives were constructed. Active full-length R·EcoRII was produced only in cells that also expressed M·EcoRII from a multicopy plasmid. Recombinant R·EcoRII bound with hybrid DNA·RNA duplexes.     

Effect of temperature and additional carbon sources on phenol degradation by an indigenous soil <Emphasis Type="Italic">Pseudomonad</Emphasis>

A new indigenous soil bacterium Pseudomonas sp. growing on phenol and on a mixture of phenol, toluene, o-cresol, naphthalene and 1,2,3-trimethylbenzene (1,2,3-TMB) was isolated and characterized. Phylogenetic analysis suggested its classification to Pseudomonadaceae family and showed 99.8% DNA sequence identity to Pseudomonas pseudoalcaligenes species. The isolate was psychrotroph, with growth temperatures ranging from ca. 0 to 40 °C. The GC–MS structural analysis of metabolic products of phenol degradation by this microorganism indicated a possible ortho cleavage pathway for high concentrations (over 200 mg L^–1) of phenol. Biodegradation rates by this species were found to be three times more effective than those previously reported by other Pseudomonas strains. The effect of temperature on phenol degradation was studied in batch cultures at temperatures ranging from 10 to 40°C and different initial phenol concentrations (up to 500mgL^–1). Above 300mgL^–1 of initial phenol concentration no considerable depletion was recorded at both 10 and 40°C. Maximum degradation rates for phenol were recorded at 30°C. The biodegradation rate of phenol was studied also in the presence of additional carbon sources (o-cresol, toluene, naphthalene, 1,2,3-TMB) at the optimum growth temperature and was found significantly lower by a factor of eight in respect to the strong competitive inhibition between the substrates and the more available sources of carbon and energy. The Haldane equation =_m S/(K_S+S+S²/K_I) was found to best fit the experimental data at the optimum temperature of 30°C than the Monod equation with kinetic constants _m=0.27h^–1, K_S=56.70mgL^–1, K_I=249.08mgL^–1.     

Improvement of growth and camptothecin yield by altering nitrogen source supply in cell suspension cultures of Camptotheca acuminata

Nitrate at 70m gave the highest biomass of Camptotheca acuminata in suspension culture in MS medium, but a NH₄ ⁺/NO₃ ^– molar ratio of 5:1 (giving a total of 40 m N) gave the maximum camptothecin yield. A two-stage flask culture system was established to improve culture efficiency; cell dry weight, camptothecin content and yield was increased by 30%, 280% and 340%, respectively when compared with those of control, reaching up to 36g l^–1, 0.36mgg^–1, and 12.8mgl^–1, respectively.     

The Effects of Hypoxia on Three Sympatric Shark Species: Physiological and Behavioral Responses

Behavioral and physiological responses to hypoxia were examined in three sympatric species of sharks: bonnethead shark Sphyrna tiburo, blacknose shark, Carcharhinus acronotus, and Florida smoothhound shark, Mustelus norrisi, using closed system respirometry. Sharks were exposed to normoxic and three levels of hypoxic conditions. Under normoxic conditions (5.5–6.4mg l^–1), shark routine swimming speed averaged 25.5 and 31.0cm s^–1 for obligate ram-ventilating S. tiburo and C. acronotus respectively, and 25.0cm s^–1 for buccal-ventilating M. norrisi. Routine oxygen consumption averaged about 234.6 mg O₂kg^–1h^–1 for S. tiburo, 437.2mg O₂kg^–1h^–1 for C. acronotus, and 161.4mg O₂ kg^–1 h^–1 for M. norrisi. For ram-ventilating sharks, mouth gape averaged 1.0cm whereas M. norrisi gillbeats averaged 56.0 beats min^–1. Swimming speeds, mouth gape, and oxygen consumption rate of S. tiburo and C. acronotus increased to a maximum of 37–39cm s^–1, 2.5–3.0cm and 496 and 599mg O₂ kg^–1 h^–1 under hypoxic conditions (2.5–3.4mg l^–1), respectively. M. norrisi decreased swimming speeds to 16cm s^–1 and oxygen consumption rate remained similar. Results support the hypothesis that obligate ram-ventilating sharks respond to hypoxia by increasing swimming speed and mouth gape while buccal-ventilating smoothhound sharks reduce activity.     

Enhancement of photorespiration in immobilized <Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis> cells

Immobilization of Chlamydomonas reinhardtii in alginate increases its photorespiration rate. In the immobilized cells, the photorespiratory enzyme, phosphoglycolate phosphatase, was 75% higher than in freely suspended cells. Thus, the immobilized cells produced glycolate at twice the rate than in freely suspended cells when treated with aminooxyacetate (a transaminase inhibitor). With immobilized cells in a batch reactor, 270mol glycolatemg^–1 Chl was produced after 12h.Revisions requested 27 October 2004; Revisions received 13 December 2004     

Effect of microrelief and vegetation on methane emission from wet polygonal tundra, Lena Delta, Northern Siberia

The effect of microrelief and vegetation on methane (CH₄) emission was investigated in a wet polygonal tundra of the Lena Delta, Northern Siberia (72.37N, 126.47E). Total and plant-mediated CH₄ fluxes were measured by closed-chamber techniques at two typical sites within a low-centred polygon. During the study period, total CH₄ flux averaged 28.0±5.4mgm^–2d^–1 in the depressed polygon centre and only 4.3±0.8mgm^–2d^–1 at the elevated polygon rim. This substantial small-scale spatial variability of CH₄ emission was caused by strong differences of hydrologic conditions within the microrelief of the polygon, which affected aeration status and organic matter content of the soils as well as the vegetation cover. Beside water table position, the vegetation cover was a major factor controlling CH₄ emission from polygonal tundra. It was shown that the dominant vascular plant of the study area, Carex aquatilis, possesses large aerenchyma, which serve as pathways for substantial plant-mediated CH₄ transport. The importance of plant-mediated CH₄ flux was strongly influenced by the position of the water table relative to the main root horizon. Plant-mediated CH₄ transport accounted for about two-thirds of the total flux in the wet polygon centre and for less than one-third of the total flux at the moist polygon rim. A clipping experiment and microscopic-anatomical studies suggested that plant-mediated CH₄ transport via C. aquatilis plants is driven only by diffusion and is limited by the high diffusion resistance of the dense root exodermes.     

Uptake of sewage derived nitrogen by<Emphasis Type="Italic"> Ulva rigida</Emphasis> (Chlorophyceae) in Bahía Nueva (Golfo Nuevo,Patagonia, Argentine)

Uptake kinetics of nitrogen derived from sewage–seawater mixtures (2.5–20% v/v effluent) were determined in the laboratory for Ulva rigida (Chlorophyceae) native from Bahía Nueva (Golfo Nuevo, Patagonia, Argentine). In terms of nitrogen concentration, experimental enrichment levels varied between 53.7 and 362.3M of ammonium and between 0.77 and 6.21M of nitrate+nitrite. Uptake rates were fitted to the Michaelis–Menten equation, with the following kinetic parameters: ammonium: V_max = 591.2molg^–1h^–1, K _s=262.3M, nitrate+nitrite: V _max=12.9molg^–1h^–1, K _s=3.5M). Both nutrients were taken up simultaneously, but ammonium incorporation was faster in all cases. The results show a high capability of Ulva rigida to remove sewage-derived nitrogen from culture media. In the field, most of the nitrogen provided by the effluent would be tied up in algal biomass, supporting low nitrogen levels found at a short distance away from the source.     

Inoculation of sugar mill by-products compost with N2-fixing bacteria

Inoculation of sugar mill by-products compost with N₂-fixing bacteria may improve its quality by increasing total N and available P. Compost was inoculated with Azotobacter vinelandii(ATCC 478), Beijerinckia derxii (ATCC 49361), and Azospirillumsp. TS8, each alone and all three together. Numbers of all N₂-fixing bacteria in compost declined from an initial population of 5×10⁵cellsg^–1 during incubation. The population of Azotobacter declined to approximately 2×10²cellsg^–1 and the population of Beijerinckia and Azospirillum declined to approximately 9×10³ and 3.5×10⁴cellsg^–1 respectively, at day 50. Inoculation with N₂-fixing bacteria increased acetylene reduction, total N by 6–16 and available P by 25–30% in comparison to the uninoculated control. Increasing the N content and P availability of compost increases its value and there may be additional benefit from providing N₂ fixing bacteria.     

Flax anther culture: effect of genotype,cold treatment and media

We report on screening of wide range of flax cultivars for androgenic response and on testing of induction conditions for flax (Linum usitatissimum L.) anther culture and plant regeneration. Anthers were cultured on four different media: Mo, N6, MS and N&N supplemented with various combinations of growth regulators. The induction of callus formation from cultured anthers was the highest on N6 (with cultivar PR FGL 77 – 12 %) and N&N media (with cultivar Carolin – 2.8 %), preferentially after cold pretreatment (7days at 8 °C). Shoots were formed on calli derived from the microspores inside the cultured anthers on media N&N and N6 supplemented with 1mgl^–1 zeatin or 1mgl^–1BAP + 1mgl^–1NAA, respectively and elongated on MS medium supplemented with 2mgl^–1 zeatin. The highest number of shoots (120) was observed with cultivar Red Wing. Shoots were rooted on MS medium supplemented with 2mgl^–1IAA. Our experiments resulted in total in 62 % anther response and 155 plants regenerated and transferred into soil.     

Increased erythritol production in Torula sp. by Mn2+and Cu2+

The production of erythritol and the erythritol yield from glucose by Torula sp. were improved, in increasing order, by supplementing with 10 mg MnSO₄4H₂O l^–1, 2 mg CuSO₄5H₂O l^–1, and both 10 mg MnSO₄4H₂O l^–1 and 2 mg CuSO₄5H₂O l^–1. Mn²⁺ decreased the intracellular concentration of erythritol, whereas Cu²⁺ increased the activity of erythrose reductase in cells. These results suggest that Mn²⁺ altered the permeability of cells, whereas Cu²⁺ increased the activity of erythrose reductase in cells.     

Larval development and food habits of the marbled parrotfish, <Emphasis Type="Italic">Leptoscarus vaigiensis</Emphasis>, associated with drifting algae

The larval development and food habits of the marbled parrotfish, Leptoscarus vaigiensis (Scaridae) associated with drifting algae were studied. In this study, 628 L. vaigiensis of various developmental stages ranging from postflexion larvae (9.4mm in standard length, SL) to adults (192.0mmSL) were sampled from drifting algae at two fishing ports in Nakagusuku Bay of Okinawa Island. In 3969 fish comprising 65 taxa in 34 families of Teleostei collected with drifting algae, L. vaigiensis occupied 15.8% of samples and occurred generally throughout the whole year. A large number of L. vaigiensis were collected from July to October accompanied by an occurrence of drifting algae composed of Sargassum spp. Larvae and early juveniles ranging from 11.1 to 14.9mmSL appeared sporadically throughout the year, and postflexion larvae 11mmSL occurred from July to November. Their food shifted from planktonic copepods in postflexion larvae and juveniles ranging from 10.0 to 14.9mmSL to seaweed in the juveniles ranging from 15.0 to 24.9mmSL. Furthermore, adults and young over 25mmSL fed almost exclusively on seaweed, with Sargassum spp. constituting the drifting algae. These facts indicate that drifting algae may have a role concerning food and habitat, and may act as a nursery for L. vaigiensis.     

First investigation of ultraoligotrophic alpine Lake Puma Yumco in the pre-Himalayas,China

Lake Puma Yumco is a typical alpine lake (altitude 5030m) located in the pre-Himalayas of Tibet, China, and this study was the first limnological investigation ever conducted on it. Lake Puma Yumco (28°34N, 90°24E) has the following morphometric properties: maximum length 31km, maximum width 14km, mean width 9km, shoreline 90km, surface area 280km², and shoreline development 1.5. Transparency was approximately 10m, even in the thawing season. The extinction coefficient of the lake water was calculated as 0.15m^–1. Annual maximum transparency was estimated from the depth of the Chara zone to be 30m. Dissolved oxygen was 7mg O₂ l^–1 and showed saturated values, and salinity was 360mgl^–1. The chemical type of the lake water was Mg-Ca-HCO₃-SO₄, and it was slightly alkaline in character. Total nitrogenous nutrients (sum of ammonia, nitrite, nitrate, and urea nitrogen), phosphate, and silicate were extremely low at 1, 0.02, and 9µM, respectively. Dissolved organic carbon, nitrogen, and phosphorus concentrations were 160, 11, and 0.08µM and the molar ratio was calculated as 2100:140:1. Chlorophyll a concentration was 0.2mgm^–3. Phytoplankton and zooplankton were dominated by Aphanocapsa sp. and Diaptomidae. Both nitrogen and phosphorus appear to be the limiting parameters for phytoplankton growth. Organic carbon and nitrogen contents in lake sediments were low and the sediments contained a large amount of CaCO₃. The grain size of sediment was that of silt-sand in most cases. The present results indicate that the pre-Himalayan alpine freshwater Lake Puma Yumco is an ultraoligotrophic lake.     

A Cytogenetic Study of the Leaf Beetle Genus Cyrtonus (Coleoptera, Chrysomelidae)

The chromosomes of ten species of Cyrtonus and the genome sizes of six are surveyed. Among the total of 15 chromosomally studied species, 11 have 2n=28 chromosomes and a 13+Xy_p male meioformula, three have 2n=40 and 19+Xy_p and one 2n=46 and 22+Xy_p. All but one species with 28 chromosomes show only metacentric or submetacentric chromosomes, whereas the species with 40 and 46 chromosomes display some telocentrics or subtelocentrics, that are probably derived from the former by centric fissions. However, since the number of major chromosome arms is strikingly higher in these latter species (NF=70 and 78) than in the 28-chromosome species (mostly NF=56), other chromosomal rearrangements such as pericentric inversions or heterochromatin accretions could also be involved. The genome sizes display a narrow range, from 1C=0.6–1.22pg, and they are not significantly correlated with the chromosome numbers. Some possible factors implied in the rough chromosomal evolution of Cyrtonus are discussed in relation to a few other genera of the subfamily Chrysomelinae.     

Gross nitrogen transformations in soils from uncut and cut boreal upland and peatland coniferous forest stands

Gross and net nitrogen (N) ammonification and nitrification were measured in soils from an uncut and recently cut upland and peatland conifer stand in northwestern Ontario, Canada. Rates of gross total inorganic N immobilization were similar to gross mineralization, resulting in low net mineralization rates in soils from all four upland and peatland conifer stands. Gross ammonification rates were variable but similar in soils from uncut and cut peatland hollows (18–19mgNkg^–1day^–1) and upland forest floor soils (14–19mgNkg^–1day^–1). Gross ammonium ( ) immobilization rates were also variable but similar to ammonification rates. Median gross nitrification rates were within 0–2mgNkg^–1day^–1 in soils from all four upland and peatland cut and uncut stands, although rates were consistently higher for the soils from the cut stands. Large variability in gross nitrification rates were observed in peatland soils, however the highest gross nitrification rates were measured in saturated peatland soils. Net rates remained low in the soils from all four stands due to high nitrate ( ) immobilization and very fast turnover (<0.2 day). Our results suggest that potential losses may be negated by high immobilization in uncut and cut boreal forest stands. This study reveals the potential for the interaction of N production and consumption processes in regulating N retention in upland and peatland conifer forests, and the resilience of the boreal forest to disturbance.     

Purification and characterization of aspartate aminotransferase from developing embryos of the camel tick Hyalomma dromedarii

Aspartate transaminase (AST) activity in the camel tick Hyalomma dromedarii was followed throughout embryogenesis. During purification of AST to homogeneity, ion exchange chromatography lead to four separate forms (termed I, II, III and IV). AST II with the highest specific activity was pure after chromatography on Sephacryl S-300. The molecular mass of AST II was 52KDa for the native enzyme, composed of one subunit of 50KDa. AST II had a Km value of 0.67mM for -ketoglutarate and 15.1mM for aspartate. AST II had a pH optimum of 7.5 with heat stability up to 50°C for 15min. The enzyme was activated by MnCl₂, and inhibited by CaCl₂, MgCl₂, NiCl₂, and ZnCl₂.