Phylogenetic identification of fungi isolated from the marine sponge Tethya aurantium and identification of their secondary metabolites

Fungi associated with the marine sponge Tethya aurantium were isolated and identified by morphological criteria and phylogenetic analyses based on internal transcribed spacer (ITS) regions. They were evaluated with regard to their secondary metabolite profiles. Among the 81 isolates which were characterized, members of 21 genera were identified. Some genera like Acremonium, Aspergillus, Fusarium, Penicillium, Phoma, and Trichoderma are quite common, but we also isolated strains belonging to genera like Botryosphaeria, Epicoccum, Parasphaeosphaeria, and Tritirachium which have rarely been reported from sponges. Members affiliated to the genera Bartalinia and Volutella as well as to a presumably new Phoma species were first isolated from a sponge in this study. On the basis of their classification, strains were selected for analysis of their ability to produce natural products. In addition to a number of known compounds, several new natural products were identified. The scopularides and sorbifuranones have been described elsewhere. We have isolated four additional substances which have not been described so far. The new metabolite cillifuranone (1) was isolated from Penicillium chrysogenum strain LF066. The structure of cillifuranone (1) was elucidated based on 1D and 2D NMR analysis and turned out to be a previously postulated intermediate in sorbifuranone biosynthesis. Only minor antibiotic bioactivities of this compound were found so far.     

Observations on different mildew sources used in apple breeding at Ahrensburg

Fifty-six populations of common bean (Phaseolus vulgaris L.) were grown in Pontevedra (Northwestern Spain) in four different environments in order to study their genetic diversity in 18 agronomical traits. All characters showed significant differences among populations, and most of them had significant genotype-environment interactions. Broad-sense heritability for this pool of characters ranged from 0.87 (seed length) to 0.12 (seed yield). Sixteen populations which deserve special attention because of their breeding value for earliness, yield, pod and seed size have been identified.     

Molecular assessment of genetic diversity in winter barley and its use in breeding

Summary During the last decades extensive progress has been achieved in winter barley breeding with respect to both, yield and resistance to fungal and viral diseases. This progress is mainly due to the efficient use of the genetic diversity present within high yielding adapted cultivars and – with respect to resistance – to the extensive evaluation of genetic resources followed by genetic analyses and introgression of respective genes by sexual recombination. Detailed knowledge on genetic diversity present on the molecular level regarding specific traits as well as on the whole genome level may enhance barley breeding today by facilitating efficient selection of parental lines and marker assisted selection procedures. In the present paper the state of the art with respect to virus diseases, i.e. Barley mild mosaic virus, Barley yellow mosaic virus, and Barley yellow dwarf virus is briefly reviewed and first results on a project aiming on a genome wide estimation of genetic diversity which in combination with data on yield and additional agronomic traits may facilitate the detection of marker trait associations and a more efficient selection of parental genotypes are presented. By field tests of 49 two-rowed and 64 six-rowed winter barley cultivars the genetic gain in yield for the period 1970–2003 was estimated at 54.6 kg ha⁻¹ year⁻¹ (r² = 0.567) for the six-rowed cultivars and at 37.5 kg ha⁻¹ year⁻¹ (r² = 0.621) for the two-rowed cultivars. Analysis of 30 SSRs revealed a non-homogenous allele distribution between two and six-rowed cultivars and changes of allele frequencies in relation to the time of release. By PCoA a separation between two and six-rowed cultivars was observed but no clear cut differentiation in relation to the time of release. In the two-rowed cultivars an increase in genetic diversity (DI) from older to newly released cultivars was detected.     

Molecular markers and doubled haploids in European plant breeding programmes

The breeding companies and laboratories involved in this article cover a wide range of crops grown in the temperate climate zone: small grain cereals, oilseed crops, forage crops, turf, vegetables and potato. Speed and efficiency are becoming increasingly important in variety breeding and doubled haploids (DH) and genetic markers are important biotechnological tools to accelerate materials to market. Collaborative research between universities, research institutions and breeding companies has resulted in the routine use of DH technology and molecular markers in practical breeding of barley, wheat and rapeseed. DH populations have been established not only for barley, wheat and rapeseed, but for rye, oat and triticale, where DH technology is less developed. A driver here is the value of the crop e.g. although wheat is less responsive to DH production the value of the end product makes the effort worthwhile. Simple and rapid DNA extraction methods used in high-throughput marker assisted selection (MAS) systems are essential for routine use of markers. MAS is used both to monitor the presence of genes of interest and also to monitor the genetic background. DH technology in forage, turf and vegetables is still in progress and the practical use of markers in all crops is limited by access to trait linked markers. Collaboration and technology transfer with universities, research institutions and breeding companies is essential for the improvement of both DH protocols in recalcitrant crops and marker technology in all crops.     

Coexpression of avian influenza virus H5 and N1 by recombinant Newcastle disease virus and the impact on immune response in chickens

To analyze the contribution of neuraminidase (NA) toward protection against avian influenza virus (AIV) infection, three different recombinant Newcastle disease viruses (NDVs) expressing hemagglutinin (HA) or NA, or both, of highly pathogenic avian influenza virus (HPAIV) were generated. The lentogenic NDV Clone 30 was used as backbone for the insertion of HA of HPAIV strain A/chicken/Vietnam/P41/05 (H5N1) and NA of HPAIV strain A/duck/Vietnam/TG24-01/05 (H5N1). The HA was inserted between the genes encoding NDV phosphoprotein (P) and matrixprotein (M), and the NA was inserted between the fusion (F) and hemagglutinin-neuraminidase protein (HN) genes, resulting in NDVH5VmPMN1FHN. Two additional recombinants were constructed carrying the HA gene between the NDV P and M genes (NDVH5VmPM) or the NA between F and HN (NDVN1FHN). All recombinants replicated well and stably expressed the HA gene, the NA gene, or both. Chickens immunized with NDVH5VmPMN1FHN or NDVH5VmPM were protected against two different HPAIV H5N1 and also against HPAIV H5N2. In contrast, immunization of chickens with NDVN1FHN induced NDV- and AIV N1-specific antibodies but did not protect the animals against a lethal dose of HPAIV H5N1. Furthermore, expression of AIV N1, in addition to AIV H5 by NDV, did not increase protection against HPAIV H5N1.     

Bactericidal activity of porcine neutrophil secretions

Antimicrobial proteins in neutrophil granules exert their bactericidal activity both within the neutrophil phagolysosome and as components of neutrophil extracellular traps. This study evaluated the bactericidal activity of porcine neutrophil secretions against four bacterial pathogens of swine. Porcine neutrophils were treated with or without phorbol myristate acetate (PMA), then the resulting supernatants were incubated with Escherichia coli K-12, Streptococcus suis, Actinobacillus suis, or Pasteurella multocida, and the surviving colony forming units were enumerated. Supernatants of PMA-activated neutrophils killed an average of 95% of E. coli K-12 cells, relative to supernatants from untreated neutrophils. Inhibition of elastase activity using chloromethylketone (CMK) prior to PMA stimulation significantly reduced the bactericidal activity of the neutrophil supernatants; 57% of the PMA-induced bactericidal activity against E. coli K-12 was estimated to be elastase-dependent. The same neutrophil supernatants had lower bactericidal activity against S. suis, A. suis, and P. multocida, with 30%, 36% and 13% reduction in bacterial numbers, respectively. The cathelicidin porcine myeloid antimicrobial peptide (PMAP)-36 and lactotransferrin were among the proteins identified in the supernatants of PMA-stimulated neutrophils by mass spectrometry. These findings imply that elastase-activated proteins, such as cathelicidins, are partially responsible for the bactericidal effect of porcine neutrophil secretions, but non-elastase-dependent proteins such as lactoferrin may also contribute. Further, the secretions of activated neutrophils were effective in killing the avirulent E. coli K-12 but were less effective against the other bacteria tested, suggesting that these pathogens may have evolved mechanisms to resist neutrophil-mediated killing.     

Einfluss differenzierter organisch-mineralischer Düngung auf die partikuläre organische Substanz des Bodens

Zusammenfassung

Im Internationalen Organischen Stickstoff-Dauerdüngungs-Versuch (IOSDV) am Standort Berlin-Dahlem wurden nach 20 Versuchsjahren chemische und physikalische Untersuchungen der organischen Bodensubstanz durchgeführt. Die langjährig differenzierte organisch-mineralische Düngung hat die Gehalte an organischem sowie heißwasserlöslichem Kohlenstoff und Gesamtstickstoff signifikant beeinflusst. Auf physikalischem Wege wurden die organischen Bestandteile des Bodens in eine leichte, mittlere und schwere Fraktion getrennt. Die Ergebnisse zeigen ein anderes Bild als die chemische Analyse. Mineralische Stickstoffdüngung und die kombinierte organische Düngung von Stroh, Rübenblatt und Zwischenfruchtgrünmasse führten im Vergleich zur ungedüngten Kontrolle zu signifikant höheren Gehalten der leichten Fraktion (LF). Stallmistdüngung beeinflusste demgegenüber die Gehalte der schweren Fraktion (SF) positiv. Stroh-, Rübenblatt- und Gründüngung erhöhten den Gehalt der schweren Fraktion nur bei gleichzeitiger mineralischer Stickstoffdüngung. Alleinige Mineralstickstoffdüngung veränderte die schwere Fraktion hingegen nicht. Mit den Fraktionen der partikulären organischen Substanz werden andere Bestandteile der organischen Bodensubstanz erfasst als bei der chemischen Humusanalyse. Somit können zusätzliche Informationen zur Beschaffenheit der organischen Bodensubstanz gewonnen werden.     

Greenhouse gas fluxes during rewetting of peatlands by use of effluents – a lysimeter study

Abstract

The present study aims at assessing the effect of using the effluent of a wastewater treatment plant as an alternative measure for rewetting nutrient-rich fen soils over the growing season on the emission of greenhouse gas (GHG) and at discussing possible changes in the greenhouse potential as a result of this practice. In order to allow a discussion on GHG based on integrated CH₄, N₂O, and CO₂ flux rates, fluxes were measured in our lysimeter study using the chamber methodology from May to December in 2003 and 2004. The study compares the gaseous fluxes of fen soils in lysimeters treated with the effluent and/or freshwater for rewetting. Only freshwater was applied to the control lysimeter. The source of water hardly had any statistically significant effect on trace gas fluxes. However, there was a trend towards higher CH₄ emissions at the effluent lysimeters compared to the control lysimeter. Effluent usage did not decrease the greenhouse effect at the same rate, which could be observed at the control. Nevertheless, regarding gaseous emissions the use of effluents could prove to be a solution to the current problem of today's major peat oxidation and fen soil loss by drainage.