Real‐time PCR detection of Erwinia amylovora on blossoms correlates with subsequent fire blight incidence          下载免费PDF全文

M. Hinze  L. Köhl  S. Kunz  S. Weißhaupt  M. Ernst  A. Schmid  R. T. Voegele 《Plant pathology》2016,65(3):462-469

Fire blight is the most devastating bacterial disease of rosaceous plants. Forecasting fire blight infections is important to allow for countermeasures that reduce economic damage in pome fruit production. Current computerized forecasting models are solely based on physical factors such as temperature and moisture, but not on the actual presence of the pathogen Erwinia amylovora. Although the inoculum concentration is considered to be crucial for infection and disease outbreak, most current approaches used for identification of fire blight inoculum including morphological, biochemical, serological, and DNA‐based methods are nonquantitative. Based on a real‐time PCR approach previously published, an improved protocol to be used directly on whole bacteria in the field is described. The method allows for early detection and quantification of the pathogen prior to the occurrence of first symptoms. There is a clear correlation between bacterial abundance and subsequent disease development. However, in some cases, no disease symptoms could be observed despite a pathogen load of up to 3·4 × 10⁶ cells per blossom. Integration of the amount of pathogen detected into refined prediction algorithms may allow for the improvement of applied forecasting models, finally permitting a better abatement of fire blight.  相似文献   

Alternative inoculum sources for fire blight: the potential role of fruit mummies and non‐host plants          下载免费PDF全文

S. Weißhaupt  L. Köhl  S. Kunz  M. Hinze  M. Ernst  A. Schmid  R. T. Voegele 《Plant pathology》2016,65(3):470-483

Fire blight is the most damaging bacterial disease in apple production worldwide. Cankers and symptomless infected shoots are known as sites for the overwintering of Erwinia amylovora, subsequently providing primary inoculum for infection in the spring. In the present work, further potential sources of inoculum were investigated. Real‐time PCR assays covering a 3‐year‐period classified 19·9% of samples taken from fruit mummies as positive. Bacterial abundance in fruit mummies during autumn, winter and spring was up to 10⁹ cells per gram of tissue and correlated well with later infection rates of blossoms. Blossoms of non‐host plants growing close to infected trees were also shown to be colonized by E. amylovora and to enable epiphytic survival and propagation of bacteria. The results indicate a potential role of fruit mummies and buds in overwintering and as a source of primary inoculum for dissemination of the pathogen early in the growing season. Non‐host blossoms may also serve as an inoculum source in the build‐up of the pathogen population. Both aspects may contribute significantly to the epidemiology of E. amylovora. The significance of infected rootstocks as an inoculum source is also discussed. Fruit mummies might be used to determine pathogen pressure in an orchard before the beginning of the blooming period.  相似文献   

Isolation,partial identification and application of diazotrophic rhizobacteria from traditional Indian rice cultivars     

Bhavanath Jha  Mukund C. Thakur  Iti Gontia  Valerie Albrecht  Marion Stoffels  Michael Schmid  Anton Hartmann 《European Journal of Soil Biology》2009,45(1):62-72

A diversity of N₂-fixing (diazotrophic) bacteria was isolated from two traditional rice cultivars, Sataria and Kartiki, from the rice growing area of Mithila region of North Bihar, India, where low levels of nitrogen fertilizers are applied. Nitrogen-free semisolid media NFb, JMV and LGI with different carbon sources and pH-values were used for enrichment and isolation of root-associated diazotrophs. The colonization density of roots by diazotrophs, as estimated from positive pellicle formation at highest dilution in nitrogen-free enrichment media, was 10⁶–10⁸ diazotrophic bacteria per g fresh root weight. Roots of the cultivar Kartiki were found to be more densely colonized endophytically by diazotrophs as detected after chloramine T (1%) surface disinfection. To ascertain the phylogenetic affiliation of the isolates, phylogenetic oligonucleotide probes and the Fluorescent in situ Hybridization (FISH) technique were applied. Using group-specific rRNA directed oligonucleotide probes, the majority of the isolates could be identified as alpha-, beta-, or gamma-proteobacteria. Using 16S and 23S rRNA-directed genus- or species-specific probes, Herbaspirillum seropedicae, Azospirillum amazonense, Burkholderia cepacia/vietnamiensis, Rhizobia and Pseudomonas spp. were found to be the most prominent root associated culturable diazotrophs. Diazotrophic Gluconacetobacter spp. were also demonstrated as colonizers of rice roots. Burkholderia cenocepacia, Pseudomonas sp. and three diazotrophic PGPR reference strains were used for the inoculation of axenically grown rice seedlings to determine the plant growth promoting potential. Significant increases in the shoot length (up to 60%), shoot dry weight (up to 33%) and the grain yield (up to 26%) per plant were observed in non-axenic pot and field trials. Using semisolid enrichment media after surface sterilization of field grown inoculated rice roots and oligonucleotide probing of the diazotrophic enrichment cultures, a sustainable colonization with the inoculated bacteria could be demonstrated.  相似文献   

Land-cover change and human population trends in the greater Serengeti ecosystem from 1984–2003     

Anna Bond Estes  Tobias Kuemmerle  Hadas Kushnir  Volker Christian Radeloff  Herman Henry Shugart 《Biological conservation》2012,147(1):255-263

The growth of human populations around protected areas accelerates land conversion and isolation, negatively impacting biodiversity and ecosystem function, and can be exacerbated by immigration. It is often assumed that immigration around protected areas is driven by attraction in the form of economic benefits, but in many cases, people may be pushed from their areas of origin toward protected areas. Mitigating the effects of immigration around protected areas necessitates understanding the actual mechanisms causing it, which can be aided by analysis of patterns of land-cover change. Our goal was to identify the reasons for human population growth and land-cover change around the protected areas in the greater Serengeti ecosystem (henceforth “the park”), and to relate agricultural conversion from 1984–2003 to trends in human demography. We found that conversion of natural habitats to agriculture was greatest closer to the park (up to 2.3% per year), coinciding with the highest rates of human population growth (3.5% per year). Agricultural conversion and population growth were greatest where there was less existing agriculture, and population density was lowest. Lack of unfarmed land farther from the park, coupled with greater poverty near the park, suggest that movement away from areas with high population densities and land scarcity was likely driving immigration near the park, where arable land was available. Our results are essential for conservation planning for one of Africa’s hallmark ecosystems, and should encourage further examination of population growth and land-cover trends near protected areas throughout the developing world.  相似文献   

Locking compression plate osteosynthesis of sacral fractures in three heifers     

Schmid TC  Kummer MR  Hagen RU  Auer JA  Nuss KA 《Veterinary surgery : VS》2011,40(3):374-378

Objective: To describe a technique used for reduction and internal fixation of sacral fractures in cattle. Study Design: Case series. Animals: Three heifers aged 19–32 months with deformation of the tail head and tail paralysis attributable to a closed fracture of S5 or S4 and S5. Methods: Fractures were reduced and repaired under general anesthesia. A narrow, 6–10‐hole, locking compression plate (LCP) was placed on the spinous processes of S5 and the first coccygeal (Co1) using 5.0 mm locking head screws. Follow‐up inquiries were made by telephone (3/3), and cows were examined clinically and radiographically 2 (1 cow) to 9 (1cow) months postoperatively. Results: No complications were encountered. The range of movement of the tail improved so that soiling of the tail during defecation was no longer a problem. In all heifers, the normal top line was restored and there was no narrowing of the pelvic canal. Unassisted calving occurred in all heifers, and follow‐up radiographs for 2 cows showed complete fracture healing. Conclusion: LCP‐osteosynthesis of the spinous processes provided sufficient stability for internal fixation of sacral fractures. Marked improvement in tail function occurred postoperatively without decompression by laminectomy.  相似文献   

GC-based analysis of plant stanyl fatty acid esters in enriched foods     

Barnsteiner A  Lubinus T  di Gianvito A  Schmid W  Engel KH 《Journal of agricultural and food chemistry》2011,59(10):5204-5214

Approaches for the capillary gas chromatographic (GC) based analysis of intact plant stanyl esters in enriched foods were developed. Reference compounds were synthesized by enzyme-catalyzed transesterifications. Their identities were confirmed by means of mass spectrometry. Using a medium polar trifluoropropylmethyl polysiloxane stationary phase, long-chain plant stanyl esters could be separated according to their stanol moieties and their fatty acid chains. Thermal degradation during GC analysis was compensated by determining response factors; calibrations were performed for ten individual plant stanyl esters. For the analysis of low-fat products (skimmed milk drinking yogurts), the GC separation was combined with a "fast extraction" under acidic conditions. For fat-based foods (margarines), online coupled LC-GC offered an elegant and efficient way to avoid time-consuming sample preparation steps. The robust and rapid methods allow conclusions on both, the stanol profiles and the fatty acid moieties, and thus provide a basis for the authentication of this type of functional food ingredients.  相似文献   

Occurrence of tympanic bone spicules and bone crests in domestic cats     

Tobias Beck  Andreas Brühschwein  Andrea Meyer-Lindenberg 《Anatomia, histologia, embryologia》2020,49(2):216-221

  相似文献   

Effect of calcareous and siliceous amendments on N2O emissions of a grassland soil     

Teodora Todorcic Vekic  Shahid Nadeem  Lars Molstad  Vegard Martinsen  Elisabeth Gautefall Hiis  Lars Bakken  Tobias Rütting  Leif Klemedtsson  Peter Dörsch 《Soil Use and Management》2023,39(3):1082-1095

Liming of acidic agricultural soils has been proposed as a strategy to mitigate nitrous oxide (N₂O) emissions, as increased soil pH reduces the N₂O/N₂ product ratio of denitrification. The capacity of different calcareous (calcite and dolomite) and siliceous minerals to increase soil pH and reduce N₂O emissions was assessed in a 2-year grassland field experiment. An associated pot experiment was conducted using homogenized field soils for controlling spatial soil variability. Nitrous oxide emissions were highly episodic with emission peaks in response to freezing–thawing and application of NPK fertilizer. Liming with dolomite caused a pH increase from 5.1 to 6.2 and reduced N₂O emissions by 30% and 60% after application of NPK fertilizer and freezing–thawing events, respectively. Over the course of the 2-year field trial, N₂O emissions were significantly lower in dolomite-limed than non-limed soil (p < .05), although this effect was variable over time. Unexpectedly, no significant reduction of N₂O emission was found in the calcite treatment, despite the largest pH increase in all tested minerals. We tentatively attribute this to increased N₂O production by overall increase in nitrogen turnover rates (both nitrification and denitrification) following rapid pH increase in the first year after liming. Siliceous materials showed little pH effect and had no significant effect on N₂O emissions probably because of their lower buffering capacity and lower cation content. In the pot experiment using soils taken from the field plots 3 years after liming and exposing them to natural freezing–thawing, both calcite (p < .01) and dolomite (p < .05) significantly reduced cumulative N₂O emission by 50% and 30%, respectively, relative to the non-limed control. These results demonstrate that the overall effect of liming is to reduce N₂O emission, although high lime doses may lead to a transiently enhanced emission.  相似文献   

Seasonal development of biomass yield in grass–legume mixtures on different soils and development of above- and belowground organs of Medicago sativa     

L. Chmelíková  S. Wolfrum  H. Schmid  M. Hejcman  K.-J. Hülsbergen 《Archives of Agronomy and Soil Science》2013,59(3):329-346

Grass–legume mixtures are suitable for crop rotations under organic farming. Little attention has been paid to seasonal development of mixtures with alfalfa under field conditions. We investigated the effects of site and cut on herbage and belowground biomass yields of grass–legume mixture and on above- and belowground traits of Medicago sativa. Six sites in southern Germany were monitored during 2011. Dry matter herbage yield ranged from 9 to 16 t ha^?1. The total herbage yield of three cuts per year decreased from 45% to 36% and 19%. The belowground biomass in the upper 30 cm soil layer ranged from 1.7 to 3.8 t ha^?1.There was no seasonal trend. Diameter of the root neck and maximum order of branching of alfalfa increased during the season. The number of nodules per plant decreased from 9.5–17.0 in May to 7.5–13.0 in August. By the last cut, roots with larger diameter created smaller nodules. More branched roots created more nodules independent of their shape. Thinner roots have more active nodules. Plant height, number of stems and inflorescences per plant were higher in July and August than in May. In conclusion, a holistic analysis including above- and belowground traits should be used for the evaluation of fodder crops.  相似文献   

Elastic moduli of living epithelial pancreatic cancer cells and their skeletonized keratin intermediate filament network     

Walter N  Busch T  Seufferlein T  Spatz JP 《Biointerphases》2011,6(2):79-85

In simple epithelia, such as living epithelial pancreatic cancer cells (Panc-1), unusual amounts of keratin filaments can be found, which makes these cells an ideal model system to study the role of keratin for cell mechanical properties. In this work, the elastic moduli of Panc-1 cells and their extracted in-situ subcellular keratin intermediate filament network are determined and compared with each other. For this, the living adherent cells and their extracted keratin network were probed with local quasistatic indentation testing during large deformations using the Atomic Force Microscope (AFM). We determined the elastic modulus of the skeletonized but structurally intact keratin network to be in the order of 10 Pa, while the living cell elastic modulus ranged from 100 to 500 Pa. By removing microfilaments, microtubules, membranes and soluble cytoplasmic components during keratin network extraction, we excluded effects caused by crosslinking with other filamentous fibers and from the viscosity of the cytoplasm. Thus, the determined elastic modulus equals the actual elastic modulus inherent to such a keratin filamentous network. In our assessment of the effective mechanical contribution of the architecturally intact, skeletonized keratin network to living cell mechanics, we come to the conclusion that it plays only a very limited role. Evidently, the quantitative dominance of keratin in these cells does not reflect a strong influence on determining the cell's elastic modulus. Instead, keratin like other filamentous structures in the cell's scaffolding, e.g., F-actin and microtubuli, is one part of a greater whole.  相似文献