A comparison of in vitro with in vivo flowering in Gentian

Young nodal explants of Gentiana triflora Pall. var. axillariflora were cultured in a woody plant medium (WPM) supplemented with B5 vitamins, sucrose (3%) and kinetin (2.0 μM). A novel observation was made in that in vitro flowering occurred following development of the axillary bud of the cultures. A comparison was made between in vitro and in vivo flowers. Although smaller in size, the in vitro flowers were morphologically comparable to the in vivo ones. Flowers from both sources were semi-opened or not opened. The colour of the in vitro flowers was paler than those in vivo. Stigma development was generally poor in both in vitro and in vivo flowers. Pollen viability was over 90% in both types of flowers. About 11% and 34% of pollen from in vitro and in vivo flowers, respectively, germinated on WPM containing 100 g l⁻¹ sucrose solidified with 10 g l⁻¹ agar. Hand pollination of stigma could raise viable seed production in in vivo-flowering plants from 0.3 o/o (i.e. without aided pollination) to 3% but none in in vitro-flowering plants where only seed-like structures, probably unfertilised ovules, were found. This revised version was published online in August 2006 with corrections to the Cover Date.     

Occurrence of Ochratoxin A-Producing Fungi in Commercial Corn Kernels in Argentina

Potentially ochratoxigenic Aspergillus and Penicillium species were identified and the natural occurrence of ochratoxin A (OTA) in corn kernels was evaluated. Likewise, the capacity to produce OTA by Aspergillus section Nigri and Circumdati was investigated. A total of 50 corn samples for human consumption was collected in the south of Córdoba Province. The surface-disinfected method for mycobiota determination was used. The OTA detection was performed by HPLC. OTA production was tested in strains belonging to section Nigri and Circumdati. Statistical analysis demonstrated that the specie A. flavus was isolated in higher frequency (p<0.01) from corn kernels in DRBC and DG18 media. The percentage of corn kernels contaminated by A. niger var. niger was similar in DRBC and DG18 media. The frequency of grains contaminated by A. flavus and A. niger var. awamori was higher than A. niger var. niger and A. japonicus var. japonicus (p<0.01) in DG18 media. The other potentially ochratoxigenic species, A. ochraceus, was isolated between 5% and 10% of the corn kernels in DG18 and DRBC media, respectively. The OTA producing species P. verrucosum was not isolated. All samples of corn were OTA negative (<1 ng g⁻¹). Thirty strains (25%) of the black Aspergillus were OTA producers. From four strains of A. ochraceus isolated, only one produced OTA. Due to the storage variable conditions could not be adequate in this substrate, the presence of ochratoxigenic strains of section Nigri and OTA needs to be evaluated for a longer time to establish the toxicological risk for human beings. The contamination of stored corn kernels with A. flavus and Aspergillus section Nigri was significant.     

Early post-fire tree regeneration in a Picea-Vaccinium forest in northern Sweden

Abstract. Age and size structure of saplings of Picea abies, Pinus sylvestris and Betula pubescens were examined in a 26-yr old forest fire area in a Picea abies-Vaccinium myrtillus forest in northern Sweden. Picea, which is a shade-tolerant species, had its maximum regeneration prior to the shade-intolerant Pinus. The shift from Picea to Pinus regeneration in the late 1970s, did not seem to be related to variations in summer temperature. Instead, it is suggested that Picea established in the shade created by dead trees, and, that increased reindeer browsing of Betula in combination with a simultaneous thinning of the tree layer, favoured Pinus recruitment. These regeneration patterns do not confirm conventional views of post-fire succession in Sweden.     

Predators of Mesoplatys ochroptera in sesbania planted-fallows in eastern Zambia

Mesoplatys ochroptera Stål (Coleoptera: Chrysomelidae) is a serious pest of the tropical legume sesbania (Sesbania sesban (L.) Merrill) widely used for soil fertility improvement in southern Africa. Surveys were conducted between October 1997 and June 1999 in order to identify the predators of M. ochroptera in sesbania fallows in eastern Zambia. The Heteroptera Afrius yolofa (Guérin-Méneville), Glypsus conspicuus (Westwood), Macrorhaphis acuta Dallas, Mecosoma mensor (Germar), Rhinocoris segmentarius (Germar), and Deraeocoris ostentans (Stål), the carabid beetle Cyaneodinodes fasciger (Chaudoir), the ants Tetramorium sericeiventre Emery and Pheidole sp., and the lacewing Mallada sp. were recorded as predators of M. ochroptera for the first time. The pentatomid bugs G. conspicuus, M. acuta and M. mensor were the most common predators in sesbania fallows. Adults and the different nymphal stages of the three species preyed on larvae, adults and occasionally on eggs of the beetle. In the insectary, the adults of G. conspicuus, M. acuta and M. mensor consumed a significantly higher number of larvae (p < 0.001) compared to adults of M. ochroptera under both free-choice and no-choice conditions. The voracity of the three species showed a positive response to prey density. The fifth instar nymphs showed variation in daily consumption of larvae, the highest number of prey being consumed two to three days after the fourth molt. These predators also attacked other pests associated with sesbania and crop plants. The potential role of the predators in the natural control of pests in agroforestry systems is discussed, with a review of their prey species.     

RTA2, a novel gene involved in azole resistance in Candida albicans

Widespread and repeated use of azoles, particularly fluconazole, has led to the rapid development of azole resistance in Candida albicans. Overexpression of CDR1, CDR2, and CaMDR1 has been reported contributing to azole resistance in C. albicans. In this study, hyper-resistant C. albicans mutant, with the above three genes deleted, was obtained by exposure to fluconazole and fluphenezine for 28 passages. Thirty-five differentially expressed genes were identified in the hyper-resistant mutant by microarray analysis; among the 13 up-regulated genes, we successfully constructed the rta2 and ipf14030 null mutants in C. albicans strain with deletions of CDR1, CDR2 and CaMDR1. Using spot dilution assay, we demonstrated that the disruption of RTA2 increased the susceptibility of C. albicans to azoles while the disruption of IPF14030 did not influence the sensitivity of C. albicans to azoles. Meanwhile, we found that ectopic overexpression of RTA2 in C. albicans strain with deletions of CDR1, CDR2 and CaMDR1 conferred resistance to azoles. RTA2 expression was found elevated in clinical azole-resistant isolates of C. albicans. In conclusion, our findings suggest that RTA2 is involved in the development of azole resistance in C. albicans.     

Variation in alcohol dehydrogenase activity in vitro in flies from natural populations ofDrosophila melanogaster

Alcohol dehydrogenase (ADH) activity variation in male flies taken directly from seven natural populations ofDrosophila melanogaster is largely accounted for by segregation of alleles at theAdh structural gene locus. There was little overlap in the ADH activities ofAdh ^F andAdh ^s homozygotes. Body weights varied only slightly betweenAdh genotypes and contributed little to ADH variation. Between and within population variation in ADH activity and ADH protein in flies in the wild is mainly due to the relative frequencies ofAdh ^F andAdh ^s.     

Displacement in two invasive species of leafminer fly in different localities

Liriomyza sativae was displaced by L. trifolii in the USA probably due to the lower insecticide susceptibility of L. trifolii, but L. trifolii was displaced by L. sativae in Japan. To know the possible causes of the species displacement observed in Japan, differences in the insecticide susceptibility, fecundity, effects of natural enemies, and reproductive interference between L. trifolii and L. sativae were evaluated. As a result, the higher fecundity of L. sativae and differential effects of parasitoids on the two Liriomyza species are considered to be contributing factors.     

Density distribution analysis of in vivo and in vitro colony forming cells in bone marrow

The technique of buoyant density separation in gradients of Bovine Serum Albumin has been used to separate hemopoietic cell populations in mouse bone marrow that form in vivo spleen colonies and in vitro colonies of granulocytes and macrophages in an agar culture system. The density distribution profiles showed a number of reproducible density subpopulations of both in vivo and in vitro colony forming cells (C.F.C.'s). The mean density of in vitro C.F.C.'s exceeded that of the in vivo but overlap of the density profiles of the two populations was evident. Density-related differences in seeding efficiency of in vivo C.F.C.'s were observed. Freund's adjuvant treatment increased marrow and spleen in vitro C.F.C. populations. Marrow density profiles obtained three and seven days after adjuvant showed a progressive increase in in vitro C.F.C.'s in a restricted density region with no associated elevation of in vivo activity. The antimitotic agent, vinblastine, revealed differences in mitotic activity between the two cell populations, reducing the in vitro C.F.C. population to .07% and the in vivo to 5% of normal in 24 hours. Density separation of vinblastine-treated marrow produced density regions devoid of in vitro activity but containing in vivo in vivo C.F.C.'s which, upon transfer to irradiated recipients, regenerated both in vivo and in vitro density distribution profiles.     

Climate-driven changes in biomass allocation in pines

Future increases in air temperature resulting from human activities may increase the water vapour pressure deficit (VPD) of the atmosphere. Understanding the responses of trees to spatial variation in VPD can strengthen our ability to predict how trees will respond to temporal changes in this important variable. Using published values, we tested the theoretical prediction that conifers decrease their investment in photosynthetic tissue (leaves) relative to water‐conducting tissue in the stem (sapwood) as VPD increases. The ratio of leaf/sapwood area (A_L/A_S) decreased significantly with increasing VPD in Pinus species but not in Abies, Pseudotsuga, Tsuga and Picea, and the average A_L/A_S was significantly lower for pines than other conifers (pines: 0.17 m² cm^?2; nonpines: 0.44 m² cm^?2). Thus, pines adjusted to increasing aridity by altering above‐ground morphology while nonpine conifers did not. The average water potential causing a 50% loss of hydraulic conductivity was ?3.28 MPa for pines and ?4.52 MPa for nonpine conifers, suggesting that pines are more vulnerable to xylem embolism than other conifers. For Pinus ponderosa the decrease in A_L/A_S with high VPD increases the capacity to provide water to foliage without escalating the risk of xylem embolism. Low A_L/A_S and plasticity in this variable may enhance drought tolerance in pines. However, lower A_L/A_S with increasing VPD and an associated shift in biomass allocation from foliage to stems suggests that pines may expend more photosynthate constructing and supporting structural mass and carry less leaf area as the climate warms.     

Undergraduate teaching in gerodontology in Austria, Switzerland and Germany

Objective: To survey the present state of undergraduate teaching in the domain of gerodontology in Germany, Switzerland and Austria. Study participants: All universities of Austria (A), Germany (D) and Switzerland (CH). Protocol: A questionnaire on undergraduate teaching in gerodontology was mailed to all Deans (A: n = 3; CH: n = 4; D: n = 31) and all independent departments except paediatric dentistry and orthodontics (A: n = 11; CH: n = 15; D: n = 111). Results: The questionnaires were completed and returned by 29 Deans (A: n = 2; CH: n = 4; D: n = 23) and 102 departments (A: n = 7; CH: n = 8; D: n = 87). In Austria, gerodontology is a very small component of the dental curriculum and the Deans did not want this to be increased. Most German universities claimed to teach some aspects of gerodontology to undergraduate students and 87.4% of the Deans voted for separate lectures in gerodontology. In Switzerland, gerodontology seems well established. The results of questionnaires from the independent departments revealed that in all three countries lectures were more prevalent (A: n = 0; CH: n = 4; D: n = 6) than practical training in nursing homes (A: n = 0; CH: n = 3; D: n = 6). Conclusion: Considering the demographical shift which is leading to an increasing proportion of elderly in the population, the weighting of gerodontology in the undergraduate dental curriculum should be considered for revision in Austria and Germany.     

Occurrence of toxigenic strains of Fusarium in maize and barley in Germany

Summary A survey was made of maize and barley in Germany for the occurrence of toxigenic strains of Fusarium and of the mycotoxins produced in culture by these strains.The following 6 species of Fusarium were found: F. avenaceum, F. culmorum, F. equiseti, F.oxysporum, F. poae, and F. tricinctum. The species most commonly isolated from bird-damaged maize ears was F. avenaceum while F. culmorum was consistently isolated from maize stem rot. The predominant species in barley grain was F. poae while F. avenaceum, F. culmorum, and F. tricinctum were also isolated frequently.Cultures on autoclaved maize of all the Fusarium strains were assayed for toxicity by feeding to 1-day-old chickens for 14 days. Some strains of F. avenaceum, F. culmorum, F. equiseti, and F. oxysporum proved to be acutely toxic to chickens and caused mortality as well as marked reductions in weight gain and feed consumption. All the strains of F. poae and F. tricinctum had a low degree of toxicity.Culture material of all the strains were analyzed for the presence of 11 known Fusarium mycotoxins. The following 4 mycotoxins were detected in the strains examined: moniliformin in 9 out of 9 F. avenaceum strains (2 to 760 ppm) and in the single strain of F. oxysporum (1150 ppm); zearalenone in 4 out of 5 F. culmorum strains (320 to 1400 ppm); deoxynivalenol in 3 out of 5 F. culmorum strains.(1 to 15 ppm); and acetyldeoxynivalenol (1 to 2 ppm) in 3 out of 5 F. culmorum strains. This is the first report of moniliformin production by F. avenaceum and F. oxysporum and also the first report of the occurrence of moniliformin-producing Fusarium strains in Europe.     

Occurrence of yeasts in psittacines droppings from captive birds in Italy

Three-hundred twenty five droppings from parrots raised in the premises of 4 breeders and in several private households were cultured for yeasts. One-hundred sixty droppings (49.2%) resulted positive. From these specimens 212 isolates belonging to 27 different species were obtained. Mainly Candida species such as C. albicans, C. catenulata, C. curvata, C. famata, C. glabrata, C. guilliermondi, C. holmii, C. intermedia, C. krusei, C. lambica, C. lusitaniae, C. membranaefaciens, C. parapsilosis, C. pelliculosa, C. sake and C. valida were isolated. Debaryomyces marama, D. polymorphus, Geotrichum sp., Pichia etchelsii, P. ohmeri, Rhodotorula glutinis, R. rubra, Rhodotorula sp., Saccharomyces cerevisiae, S. kluyiveri and Zygosaccharomyces sp. were also obtained. Dark colonies on Staib medium were never observed. The psittacine birds apparently serve as carriers for several Candida species or their perfect states and to a lesser extent for other opportunistic yeasts such as Rhodotorula, Trichosporon and Saccharomyces spp., which are considered part of the transient microbiota of the gastrointestinal tract. The most striking finding was the absence of Cryptococcus spp. among the isolates. The present survey confirms the role of pet birds in carrying potential zoonotic yeasts. This revised version was published online in June 2006 with corrections to the Cover Date.     

A nonsense mutation in a putative sulphate transporter gene results in low phytic acid in barley

Low phytic acid grains can provide a solution to dietary micronutrient deficiency and environmental pollution. A low phytic acid 1-1 (lpa1-1) barley mutant was identified using forward genetics and the mutant gene was mapped to chromosome 2HL. Comparative genomic analysis revealed that the lpa1-1 gene was located in the syntenic region of the rice Os-lpa-MH86-1 gene on chromosome 4. The gene ortholog of rice Os-lpa-MH86-1 (designated as HvST) was isolated from barley using polymerase chain reaction and mapped to chromosome 2HL in a doubled haploid population of Clipper×Sahara. The results demonstrate the collinearity between the rice Os-lpa-MH86-1 gene and the barley lpa1-1 region. Sequence analysis of HvST revealed a single base pair substitution (C→T transition) in the last exon of the gene in lpa1-1 (M422), which resulted in a nonsense mutation. These results will facilitate our understanding of the molecular mechanisms controlling the low phytic acid phenotype and assist in the development of a diagnostic marker for the selection of the lpa1-1 gene in barley.     

Distribution of cephalopods in the upper epipelagic northwestern Bering Sea in autumn

In the northwestern Bering Sea in autumn, the epipelagic cephalopod community was represented by the boreal fauna, and was found to be composed of three families and nine species of the order Teuthida: Gonatidae (Berryteuthis magister, Boreoteuthis borealis, Gonatopsis japonicus, Gonatus madokai, Gonatus kamtschaticus, Gonatus onyx, and Gonatus pyros), Chiroteuthidae (Chiroteuthis calyx) and Onychoteuthidae (Onychoteuthis borealijaponica). Two pelagic gonatid species (B. borealis and G. kamtschaticus) dominated the cephalopod community in the upper 50 m. The distribution patterns of B. borealis and G. onyx were associated with diel vertical migrations of these squid. The distribution of two distinct size groups of G. kamtschaticus suggested ontogenetic migration of larger squid to deeper layers, and adds to previous data suggesting that this species may be a heterogeneous assemblage. Demersal B. magister rarely occurred in the surface waters. The occurrence of maturing O. borealijaponica in the southern marine area indicated that these were occasional seasonal migrants from the ocean. The occurrence of juvenile C. calyx suggested that these squid may conduct vertical forage migrations from deep waters to the surface layers.     

Predation on snails by an indigenous fish,Sargochromis codringtonii,in ponds in Zimbabwe

The extent to which Sargochromis codringtonii (a known predator of snails) can control snail numbers was investigated in cementlined ponds in order further to evaluate the fish's potential as an agent for the biological control of fresh water snails, especially Bulinus globosus which is the intermediate host for Schistosoma haematobium in Zimbabwe. Bulinus tropicus of all sizes were vulnerable to predation by fish, but both Melanoides tuberculata and Bulinus globosus with shell heights greater than 10mm seemed to be less so. Surprisingly, the density of B. globosus was lower in ponds without fish than in ponds with fish, while the density of B. tropicus was very high in ponds without fish. It was not possible to conclude whether B. globosus benefited by the presence of fish, or was less palatable to the fish than B. tropicus, or whether the high density of B. tropicus in ponds without fish exerted competitive effects on B. globosus. Consequently, further experiments are required to determine whether S. codringtonii can reduce populations of B. globosus in the absence of B. tropicus.     

Detection of European Peristenus digoneutis loan in mirid populations in southern Ontario

Lygus spp. Hahn (Hemiptera: Miridae) are serious pests of a wide variety of economically important crops in North America. A European parasitoid, Peristenus digoneutis Loan (Hymenoptera: Braconidae), released in the northeastern USA for the biological control of Lygus, has successfully established in nine states and in eastern Canada, including southeastern Ontario, southern Quebec and Nova Scotia. To determine the extent to which P. digoneutis has dispersed and established in mirid populations in Ontario, a single-step multiplex PCR assay, designed to differentiate European species P. digoneutis and P. relictus from native North American Peristenus species, was used to identify parasitoid larvae dissected from field-collected Miridae. A total of 222 parasitoid larvae were analysed with the multiplex assay. Most (172) were identified as members of the P. pallipes Curtis complex; however, three from the Niagara region were identified as P. digoneutis. Specimens that did not amplify using the Peristenus multiplex PCR assay were screened with a PCR primer set designed to detect Leiophron (Hymenoptera: Braconidae) species, and most were confirmed to be Leiophron larvae. This study confirms the presence of P. digoneutis species in southern Ontario, and is another example of the utility of molecular methods for the detection of newly introduced or dispersed parasitoids. The presence of exotic P. digoneutis in southern Ontario may expedite future releases to augment already-established populations. Following such releases, the Peristenus multiplex PCR assay will be a useful component of post-release studies to evaluate the success of the biological control programme for Lygus plant bugs in Canada.     

Neisseria gonorrhoeae acquires mutations in analogous regions of gyrA and parC in fluoroquinolone-resistant isolates

Neisseria gonorrhoeae homologues of gyrA and parC have been identified using hybridization probes generated from conserved regions of diverse gyrA genes. These genes have been tentatively identified as gyrA and parC, based on predicted amino acid sequence homologies to known GyrA homologues from numerous bacterial species and to ParC from Escherichia coli and Salmonella typhimurium. The gyrA gene maps to a physical location distant from the gyrB locus on the gonococcal chromosome, which is similar to the situation found in E. coli. The parC gene is not closely linked (i.e. greater than 9 kb) to an identifiable parE gene in N. gonorrhoeae. The gonococcal GyrA is slightly larger than its E. coli homologue and contains several small insertions near the O-terminus of the predicted open reading frame. A series of ciprofloxacin-resistant mutants were selected by passage of N. gonorrhoeae on increasing concentrations of the antibiotic. Sequential passage resulted in the selection of isolates with minimum inhibitory concentrations approximately 10000-fold higher than the parental strain. Mutations within gyrA resulted in low to moderate levels of resistance, while strains with high-level resistance acquired analogous mutations in both gyrA and parC. Resistance mutations were readily transferred between N. gonorrhoeae strains by transformation. The frequencies of transformation, resulting in different levels of ciprofloxacin resistance, further support the notion that both gyrA and parC genes are invoived in the establishment of extreme levels of ciprofloxacin resistance.     

Recent advances in Pelargonium in vitro regeneration systems

Recent advances in the development of protocols for in vitro culture and genetic manipulation have provided new avenues for the development of novel varieties of Pelargonium and for use as model systems for investigating the factors controlling plant morphogenesis. Optimized techniques of meristem culture have supplemented the culture indexing methods in commercial greenhouse production resulting in availability of large-scale pathogen indexed planting material. Currently, technologies are available for the mass in vitro propagation of F1 hybrid Pelargonium through both organogenesis and somatic embryogenesis. The somatic embryogenesis model system has allowed researchers to identify critical factors controlling plant morphogenesis in vitro such as regulation of regeneration by growth regulators, choice of explant and characterization of induction and expression phases of morphogenesis in Pelargonium. Also, optimization of technologies for genetic transformation of Pelargonium opened up the possibilities for developing genotypes with novel characters, including resistance to some of the major diseases. Finally, the development of regeneration systems for Pelargonium spp. has facilitated conventional crop improvement programs, thereby providing a valuable resource to the horticultural industry.     

Lack of intraspecific variation in cpDNA in Trichocolea tomentella

Abstract

The Asian genus Struckia Müll.Hal. is reduced to a synonym of Plagiothecium Bruch &; Schimp. according to a phylogenetic analysis involving S. argentata (Mitt.) Müll.Hal., S. enervis (Broth.) Ignatov, T.J.Kop. &; D.G.Long and 13 representative boreal species of Plagiothecium. Two nuclear regions, ITS and partial gapC, and five chloroplast regions, rbcL, rps4-trnS, psaB, trnG and trnL-F, were utilized to reconstruct the phylogenetic relationship between Struckia and Plagiothecium. Bayesian, maximum parsimony, and maximum likelihood analyses resulted in a strongly supported clade including Struckia and Plagiothecium. Plagiothecium handelii Broth. and P. paleaceum (Mitt.) A.Jaeger, which share similar geographical ranges with Struckia and intermediate morphological traits between Struckia and Plagiothecium, grouped with the Chinese Struckia, and all of them appeared as sister to the ‘core’ of Plagiothecium. The accurate position of P. piliferum (Sw.) Schimp. remains unclear due to moderate support from all analyses, and we suggest that it is retained in Plagiothecium until further evidence is available.