REMI (Restriction Enzyme Mediated Integration) and its Impact on the Isolation of Pathogenicity Genes in Fungi Attacking Plants

Development of molecular techniques for phytopathogenic fungi aims at the identification of fungal genes whose products are essential for successful infection of the host plant. Initial approaches have relied on isolating candidate genes and generating null-mutations by homologous recombination. Unfortunately, the results of this strategy have not been overly successful. This has led to a search for alternatives which allow an unbiased identification of pathogenicity genes. One method, which has proved successful in several systems, is a tagging mutagenesis procedure termed restriction enzyme mediated integration (REMI). In this mini-review we describe this procedure and review its features and results of its use when applied to the identification of fungal genes required for disease development in planta.     

Niche‐based species distribution models and conservation planning for endangered freshwater crayfish in south‐western Germany

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RADIOGRAPHIC IMAGING OF OTITIS MEDIA AND INTERNA IN PIGS

Middle and inner ear infections have been reported as a clinical entity in swine, other animal species and humans. In pigs, the anatomical-pathological and microbiological findings have been described. In this report, we describe radiographic findings in affected pigs. A total of 25 pigs with a head tilt and circling, as clinical signs of otitis media and interna, were examined. The majority were weaner-pigs with dyspnea or rhinitis. In radiographs, there was an increased opacity of the bulla tympanica, often accompanied by marginal destruction or thickening of the bulla wall. The radiographic findings confirmed the clinical diagnosis in each affected pig, but there were 5 false positive interpretations.     

Health screening of free-ranging European brown hares (Lepus europaeus) on the German North-Sea island Pellworm

Background

A sudden decline of the European brown hare (Lepus europaeus) population in one of the best hunting districts for small game species in northern Germany, the German North-Sea island Pellworm, in the years 2007/08 following marked habitat changes led to the implementation of a thorough health assessment program of the population. 110 animals were collected during the normal hunting season in the years 2010 and 2011. A post-mortem examination and histopathological investigation was performed on all animals. Additionally, routine bacteriology of the small intestine and parasitology were carried out. Sera of hares were tested for European Brown Hare Syndrome (EBHS) by enzyme linked immunosorbent assay, and for Treponema sp. by indirect immunofluorescent test. Additional testing was performed when deemed necessary.

Results

The most striking result was a shift in the intestinal bacterial flora towards Gram-negative Enterobacteriaceae with a predominance of either Escherichia coli, or Aeromonas sp., or a high-grade double-infection with these two pathogens with subsequent catarrhal enteritis. Additionally, a marked coccidiosis, and varying infestations with the nematode Trichostrongylus retortaeformis were found. The sero-prevalence for EBHS was 78.1%, and for Treponema 43.9%.

Conclusions

The shift and decrease in diversity of the intestinal flora was the main and most consistent result found. In the authors’ opinion the change of the habitat combined with other stressors increased the animals’ sensitivity to ubiquitous bacterial species and parasites which usually would not have such fatal effects.     

Effects of the nitrification inhibitor nitrapyrin and the plant growth regulator gibberellic acid on yield-scale nitrous oxide emission in maize fields under hot climatic conditions

Nitrification inhibitors are widely used in agriculture to mitigate nitrous oxide(N₂O)emission and increase crop yield.However,no concrete information on their mitigation of N₂O emission is available under soil and environmental conditions as in Pakistan.A field experiment was established using a silt clay loam soil from Peshawar,Pakistan,to study the effect of urea applied in combination with a nitrification inhibitor,nitrapyrin(2-chloro-6-tri-chloromethyl pyridine),and/or a plant growth regulator,gibberellic acid(GA_3),on N₂O emission and the nitrogen(N)uptake efficiency of maize.The experimental design was a randomized complete block with five treatments in four replicates:control with no N(CK),urea(200 kg N ha^-1)alone,urea in combination with nitrapyrin(700 g ha^-1),urea in combination with GA_3(60 g ha^-1),and urea in combination with nitrapyrin and GA_3.The N₂O emission,yield,N response efficiency,and total N uptake were measured during the experimental period.The treatment with urea and nitrapyrin reduced total N₂O emission by 39%–43%and decreased yield-scaled N₂O emission by 47%–52%,relative to the treatment with urea alone.The maize plant biomass,grain yield,and total N uptake increased significantly by 23%,17%,and 15%,respectively,in the treatment with urea and nitrapyrin,relative to the treatment with urea alone,which was possibly due to N saving,lower N loss,and increased N uptake in the form of ammonium;they were further enhanced in the treatment with urea,nitrapyrin,and GA_3 by 27%,36%,and 25%,respectively,probably because of the stimulating effect of GA_3 on plant growth and development and the reduction in biotic and abiotic stresses.These results suggest that applying urea in combination with nitrapyrin and GA_3 has the potential to mitigate N₂O emission,improve N response efficiency,and increase maize yield.     

Nitrification inhibitor nitrapyrin does not affect yield-scaled nitrous oxide emissions in a tropical grassland

Urea is the most common nitrogen(N)fertilizer used in the tropics but it has the risk of high gaseous nitrogen(N)losses.Use of nitrification inhibitor has been suggested as a potential mitigation measure for gaseous N losses in N fertilizer-applied fields.In a field trial on a tropical Andosol pastureland in Costa Rica,gaseous emissions of ammonia(NH₃)and nitrous oxide(N₂O)and grass yield were quantified from plots treated with urea(U;41.7 kg N ha^-1application^-1)and urea plus the nitrification inhibitor nitrapyrin(U+NI;41.7 kg N ha^-1application^-1and 350 g of nitrapyrin for each 100 kg of N applied)and control plots(without U and NI)over a six-month period(rainy season).Volatilization of NH₃(August to November)in U(7.4%±1.3%of N applied)and U+NI(8.1%±0.9%of N applied)were not significantly different(P>0.05).Emissions of N₂O in U and U+NI from June to November were significantly different(P<0.05)only in October,when N₂O emission in U+NI was higher than that in U.Yield and crude protein production of grass were significantly higher(P<0.05)in U and U+NI than in the control plots,but they were not significantly different between U and U+NI.There was no significant difference in yield-scaled N₂O emission between U(0.31±0.10 g N kg^-1dry matter)and U+NI(0.47±0.10 g N kg^-1dry matter).The results suggest that nitrapyrin is not a viable mitigation option for gaseous N losses under typical N fertilizer application practices of pasturelands at the study site.     

Quantifying moisture and roughness with Support Vector Machines improves spectroscopic soil organic carbon prediction

The challenges of Vis‐NIR spectroscopy are permanent soil surface variations of moisture and roughness. Both disturbance factors reduce the prediction accuracy of soil organic carbon (SOC) significantly. For improved SOC prediction, both disturbance effects have to be determined from Vis‐NIR spectra, which is especially challenging for roughness. Thus, an approach for roughness quantification under varying moisture and its impact on SOC assessment using Support Vector Machines is presented here.     

Erosion processes and erodibility of cultivated soils in North Rhine‐Westphalia under artificial rain. II. Results of field experiments and comparison with laboratory trials

In the densely populated state of North Rhine‐Westphalia, soil erosion by water causes substantial on‐site degradation and off‐site damages. The implementation of soil‐conservation measures is improved, if soil erodibilities and erosion processes are known. In a state‐wide investigation, we aimed to representatively assess soil‐erosion processes and erodibilities of cultivated soils. For this purpose, we measured surface runoff and soil‐loss rates of 28 cultivated soils with field plots under artificial rain. In the field experiments, surface runoff and soil loss indicated high sealing susceptibilities and high erodibilities on soils of quite different textures including a clay silt, a loam silt, a loam sand, a sand loam, and two standard loams. Rill formation causing high soil‐loss rates was observed on a clay silt (soil BM) and on a loam silt (soil RB), the latter yielding an empirical K‐factor of 1.66 t ha^–1 h N^–1. K‐factors of other silty soils ranged from 0.04 to 0.48, whereas sandy soils and clayey soils had K‐factors ranging from 0.00 to 0.32, and 0.00 to 0.12, respectively. Comparatively high erodibilities of two silt clays were due to saturation overland flow. Erosion processes and erodibilities of soils with similar texture varied to a large extent, possibly caused in part by seasonal differences in the timing of erosion tests. Surface runoff was different in field experiments compared with laboratory experiments (companion paper) conducted with topsoil material taken from the field plots. In addition, higher concentrations of suspended sediment were recorded on average in the field than in the laboratory. These differences might reflect the influence of the subsoil and are due to higher transport capacities on longer plots in the field. Thus, laboratory experiments can complement but not replace costly field trials for K‐factor determination. Empirical K‐factors derived from field and laboratory experiments are in general lower than K‐factors of other soils in Germany or calculated K‐factors derived from pedotransfer functions, which might be attributed to a more maritime‐type climate in North Rhine‐Westphalia. Since the temporal variability of erodibility was not assessed, the reported K‐factors should be regarded as preliminary.