Design and evaluation of nematode 18S rDNA primers for PCR and denaturing gradient gel electrophoresis (DGGE) of soil community DNA

Consensus nematode 18S ribosomal DNA primers were designed by aligning available 18S sequences and identifying a variable region flanked by highly conserved regions. These primers were then used to amplify nematode 18S rDNA from whole soil community DNA extracted from a range of European grassland types. Cloning of the PCR amplicons (778 bp) followed by restriction digest analysis (RFLP) resulted in the recovery of 34 unique nematode sequences from the four grasslands studied. Comparison of these data with the limited number of 18S rDNA nematode sequences currently held in on-line databases revealed that all of the sequences could be assigned to known nematode taxa albeit tentatively in some cases. Two of the sequences recovered from the site in the Netherlands (wet, hay-grassland) were recovered in a clade that included a sequence of the genus Trichodorus whilst other sequences from this site showed similarity with 18S rDNA sequences of the genus Prismatolaimus (five sequences), Xiphinema (one sequence) and Enoplus (one sequence). Of the remaining sequences, two showed some affinity with Mylonchulus (UK, upland peat), four with Steinernema (UK) and one sequence with Mesorhabditis (Hungary, east European Steppe). Three sequences from the Netherlands and one from Hungary were recovered in a clade that included a sequence of the genus Pratylenchoides whilst three further sequences from the Netherlands and two from Hungary were recovered in a clade encompassing the genus Globodera. Of the remaining nine sequences, two (NL6, NL62) formed a distinct lineage within the Adenophorea with 90% bootstrap recovery in a paraphyletic clade that included sequences of Prismatolaimus and Trichodorus. Seven sequences (three from the Netherlands, three from the UK and one from Greece) were left unassigned though the tree topology suggested some relationship (58% bootstrap recovery) with the genus Cephalobus. To assess whether primers used to amplify 18S rDNA might be used to fingerprint genetic diversity in nematode communities in soil, the environmental sequence data were used to design a second set of primers carrying a GC-clamp. These primers amplified a 469 bp fragment internal to the region flanked by the primer set used to derive the nematode trees and were used to amplify 18S rDNA for subsequent analysis using denaturing gradient gel electrophoresis (DGGE). DGGE analysis of six major European grassland types revealed considerable genetic diversity between sites. However, the relationships seen with the DGGE data were inconsistent with previous studies where the same soils had been characterized with respect to functional and morphological diversity. To confirm that this second set of primers was amplifying nematode sequences, selected bands on the DGGE gels were extracted, PCR amplified and sequenced. The final alignment was 337 bases. These analyses revealed the presence of sequence signatures from the genera Paratrichodorus, Plectus, Steinernema, Globodera, Cephalobus and Pratylenchoides.     

Landscape-moderated importance of hedges in conserving farmland bird diversity of organic vs. conventional croplands and grasslands

Hedges provide important nesting, feeding and sheltering sites for birds in agricultural areas, while organic farming also enhances farmland birds. However, it is little known how the interaction of these local variables (amount of hedges and organic management) with landscape scale variables affects birds. We selected paired conventional and organic winter wheat fields and meadows in each of 10 landscapes in Germany. Birds were surveyed in the fields and in the adjoining hedges. More bird species occurred in organic than in conventional fields regardless of land-use type (wheat fields and meadows). However, hedge length had a much stronger effect on bird richness than organic farmland management. We found an interaction of landscape complexity and hedge length in that hedge length enhanced bird richness only in case of simple landscapes (<17% semi-natural areas within a 500 m radius around the centre of bird survey plots). In more complex landscapes the local effect of hedge length levelled off because bird richness was high even without local hedges. Therefore, adding hedges or introducing organic farming practices should be primarily promoted in simple landscapes, where it really makes a difference for biodiversity.     

Local early induced resistance of plants as the first line of defence against bacteria

This paper is an overview of a non-specific local early induced resistance (EIR) mechanism, distinct from the incompatible-specific hypersensitive reaction (HR). We have shown that the local induced resistance (LIR) described earlier is not a single and uniform response to pathogen infection, because an early (EIR) and a late form can be distinguished. EIR operates from 3-6 h post-inoculation (hpi) until about 20 hpi, and is inhibited by a short heat-shock or the eukaryotic protein synthesis inhibitor, cycloheximide. In contrast, LIR, which corresponds to the induced resistance forms discovered earlier, requires more time (about 24 h) and intensive illumination to develop, and is effective for a longer period. EIR develops parallel with HR and is sometimes able to prevent it when the induction time of HR is longer than the time required for the development of EIR. It seems that EIR inhibits the metabolism of bacteria and the activity of hrp genes which otherwise are required for the induction of HR. In a compatible host-pathogen relationship the effect of EIR fails to take place. The rapid development of EIR is greatly influenced by temperature and the physiological state of the plant. EIR activates the accumulation of hydrogen peroxide at the bacterial attachment, expressing new peroxidase isoenzymes in the initiated plant tissue. It seems that this is a native general local defence mechanism which can localise foreign organisms even at the penetration site.     

NORMAL DUPLEX DOPPLER WAVEFORMS OF MAJOR ABDOMINAL BLOOD VESSELS IN DOGS: A REVIEW

The normal flow velocity profile and duplex Doppler waveform of the major abdominal blood vessels (aorta, caudal vena cava and the portal vein as well as their major branches) were examined by Doppler ultrasound. The flow velocity profile of an artery is largely determined by its diameter. The pulsatility of the waveform is related to the vascular impedance downstream to the point of measurement. Early systolic peak is present in the Doppler pattern of some vessels in some dogs. The waveform of the veins is mainly affected by the pressure conditions of the right atrium and the intrathoracic and intraabdominal pressure changes due to the respiration. Simultaneous electrocardiogram was used to reveal the effect of the heart beats on the Doppler patterns of the veins.     

Characterisation of Pasteurella multocida isolated from fowl cholera outbreaks on turkey farms

SUMMARY: Biochemical profiles, restriction endonuclease analysis (REA) and ribotyping were used to investigate Pasteurella multocida isolates from outbreaks of fowl cholera on 7 turkey farms in New South Wales. While only a single isolate was available from 5 of the farms, multiple isolates, 4 and 12 respectively, were available from the other 2 farms. The available field evidence suggested that 8 outbreaks had occurred with one farm suffering 2 outbreaks. The isolates obtained were all confirmed as Pasteurella multocida . Biochemical profiles allocated the isolates to 4 groups, 3 being variants of P multocida subsp multocida and the fourth being P multocida subsp septica . REA performed with Hpall established 7 groups. Ribotyping using the Hpall digests probed with the 16S rRNA operon of Haemophilus paragallinarum recognised the same 7 groups as REA. Unlike the biochemical profiles, both REA and ribotyping provided a fine subdivision that identified outbreaks as either related or unrelated. The REA and ribotyping patterns as well as biochemical profiles were stable for all isolates from the outbreaks in which multiple isolates were obtained from either the same bird or from different birds. REA and ribotyping were found to be superior to biotyping methods for the investigation of fowl cholera outbreaks.     

Model legumes contribute to faba bean breeding

Faba bean is an excellent candidate crop to provide nitrogen input into temperate agricultural systems. However, its growth is hampered by several factors including environmental stresses and the presence of anti-nutritional factors. To solve these limitations, breeding programs have been initiated that were successful for monogenic traits but not so for multigenic traits. The large genome size of faba bean has slowed down breeding processes. Several other legumes have emerged as model legumes including Medicago truncatula, Lotus japonicus, Glycine max and Pisum sativum. The establishment of these models has already boosted our understanding of important processes such as the nitrogen-fixing symbiotic interaction. The high level of synteny and collinearity existing between legumes makes possible the transfer of key knowledge from model legumes to faba bean. Here we review the most recent knowledge gained from model legumes on grain quality, resistance to biotic and abiotic stresses, nitrogen-fixing symbiosis and how this knowledge can be employed for faba bean breeding.     

Crown plasticity and neighborhood interactions of European beech (Fagus sylvatica L.) in an old-growth forest

Competition for canopy space is a process of major importance in forest dynamics. Although virgin and old-growth European beech (Fagus sylvatica L.) forests in Europe have been studied for many years, there are to date no studies of individual-tree crown plasticity and the way this is influenced by local neighborhood interactions in these forests. In this study, we analyzed crown plasticity and local neighborhood interactions of individual trees in the upper canopy of the old-growth beech forests of Serrahn, northeast Germany. In a 2.8-ha sample plot, we measured crown radii of all upper canopy trees and analyzed the direction and extent of crown asymmetry. Size, relative position, and distance of neighboring trees were used to construct vectors of neighborhood asymmetry within different distances from target trees. The crowns of beech trees showed strong morphological plasticity. Mean absolute and relative displacement of crown centers from the stem base were 1.95 m and 0.37, respectively. Circular–circular rank correlation coefficients between the direction of crown displacement and the direction of neighborhood pressure showed that trees strongly positioned their crowns away from local neighbors. Highest correlation coefficients were obtained when basal area and relative position of neighboring trees within a radial distance of 12 m were considered. Clark and Evans index and Ripley’s K-function showed that crowns were more regularly distributed than stems. Projected canopy cover was about 10% higher than canopy cover with simulated circular crowns. We conclude that the crowns of older beech trees have a high ability to plastically respond to changes in the local canopy conditions, enabling very effective exploitation of canopy space.     

Evaluation of kaolin and cinnamon essential oil to manage two pests and a fungal disease of sour cherry at different tree canopy levels

Journal of Plant Diseases and Protection - We studied the effects of kaolin particle film applications on major sour cherry (Prunus cerasus L.) pests and fruit quality and evaluated the ability of...