Untersuchungen zur Übertragung von Clavibacter michiganensis ssp. sepedonicus auf gesunde Kartoffelknollen

Zusammenfassung In den Jahren 1999 bis 2003 wurde in Freiland-, Klimakammer- und Lagerungsversuchen überprüft, ob ein Risiko für die Übertragung des Erregers der Bakteriellen Ringfäule der Kartoffel (Clavibacter michiganensis ssp. sepedonicus) besteht, wenn (a) gesunde Kartoffelknollen in Kontakt mit Maschinen und Geräten kommen, die mit dem Erreger kontaminiert sind (indirekter Kontakt) und (b) gesunde Kartoffelknollen direkt in Kontakt mit infizierten Knollen kommen (direkter Kontakt). Nach indirektem Kontakt konnte nur beim nachfolgenden Anbau der kontaminierten Knollen in der Klimakammer Befall in Kraut und Knollen festgestellt werden. Im Freiland konnte der Erreger, auch bei wiederholtem Nachbau der geernteten Knollen, nicht nachgewiesen werden. Nach direktem Kontakt und nachfolgendem Anbau der kontaminierten Knollen in der Klimakammer und im Freiland, wurde der Erreger in allen Fällen in den geerntete Knollen nachgewiesen. Befall im Kraut wurde nur in dem Klimakammerversuch und in einem Freilandversuch ermittelt. Wurden durch direkten Kontakt kontaminierte Knollen eingelagert, konnte der Erreger in allen untersuchten Knollen festgestellt werden. Insgesamt besteht ein hohes Risiko, dass gesunde Knollen infiziert werden, wenn oberflächliche Kontaminationen mit dem Erreger erfolgen. Die Wahrscheinlichkeit von Infektionen steigt mit zunehmender Kontaminationsstärke.     

Überleben von Ralstonia solanacearum Biovar 2 (Rasse 3), dem Erreger der Schleimkrankheit der Kartoffel, in Boden und auf verschiedenen Materialien (Mikrokosmos)

Microcosm studies were carried out to test the survival of Ralstonia solanacearum biovar 2 (race 3) in soil at the permanent wilting point (wp) water content and at field capacity (fc) water content and on various material. Soils were placed at permanent ?5°C, 4°C, 15°C and 20°C and weekly fluctuating ?10/0/+10°C and the material at 5, 15 °C, 20°C with relative humidity (rh) uncontrolled or at constant 10% or 90%. In soil, survival was clearly dependent on temperature independent of water content. At 20°C Ralstonia solanacearum could be reisolated up to 364 days, at 15°C up to 290 days, at 4°C up to 209 days and at fluctuating temperatures (?10/0/+10°C) only up to 18 days. The lower the temperature, the more the population declined. At 15°C and 20°C appr. 10⁷ cfu/g soil were detected after 100 days, whereas at ?5°C only 10² cfu/g soil were detected after only 18 days. The pathogen was longer detectable in sandy-clay loam than in lighter sandy soil. It could be longer reisolated at wilting point and the populations did not decline as rapidly as at field capacity. Ralstonia solanacearum could best survive on material surfaces like rubber, plastic and varnished metal with maximum survival of 40 days at 5°C and 10% rh. In general there is a low risk of Ralstonia solanacearum overwintering under European climatic conditions when the fields are cleared of plant debris and the soil is frozen. Contamined material surfaces pose the risk of pathogen transmission to healthy tubers.     

Short-term storage and cryopreservation of milt from Atlantic salmon and sea trout

Experiments on short-term preservation of sperm were performed with Atlantic salmon (Salmo salar). Fertility was maintained for up to 10 days when 2 mm thick samples were stored at 0° C under an oxygen atmosphere in the presence of antibiotics (125 IU penicillin and 125 μg streptomycin per ml sperm). Fertility was completely lost after 24 days. Sperm stored without antibiotics fertilized 100% of eggs after 6 days.

Cryopreservation was carried out with milt from Atlantic salmon and sea trout (Salmo trutta). Semen mixed with extender was frozen on dry ice (pellets) with subsequent storage in liquid nitrogen. Sperm pellets were thawed in a 0.12-M NaHCO₃-solution (10° C) before insemination. The suitability of an extender as previously described by Stoss and Holtz and of a 0.3-M glucose solution with the addition of 10% DMSO, was tested on two different batches of sperm and eggs in Atlantic salmon and sea trout. In addition, the extender earlier reported by Mounib and an aqueous solution of 10% DMSO were only used in Atlantic salmon with one batch of gametes.

Insemination with cryopreserved Atlantic salmon sperm resulted in 36 to 91.3% eyed eggs (control = 100). The differences were caused by the type of extender and the batch of gametes employed. The very simple extender consisting of 0.3 M glucose and 10% DMSO only was the most successful. Results with cryopreserved sea trout sperm ranged between 38.6–54.8% eyed eggs, showing no difference between treatments.     

Camel Streptococcus agalactiae populations are associated with specific disease complexes and acquired the tetracycline resistance gene tetM via a Tn916-like element

Camels are the most valuable livestock species in the Horn of Africa and play a pivotal role in the nutritional sustainability for millions of people. Their health status is therefore of utmost importance for the people living in this region. Streptococcus agalactiae, a Group B Streptococcus (GBS), is an important camel pathogen. Here we present the first epidemiological study based on genetic and phenotypic data from African camel derived GBS. Ninety-two GBS were characterized using multilocus sequence typing (MLST), capsular polysaccharide typing and in vitro antimicrobial susceptibility testing. We analysed the GBS using Bayesian linkage, phylogenetic and minimum spanning tree analyses and compared them with human GBS from East Africa in order to investigate the level of genetic exchange between GBS populations in the region. Camel GBS sequence types (STs) were distinct from other STs reported so far. We mapped specific STs and capsular types to major disease complexes caused by GBS. Widespread resistance (34%) to tetracycline was associated with acquisition of the tetM gene that is carried on a Tn916-like element, and observed primarily among GBS isolated from mastitis. The presence of tetM within different MLST clades suggests acquisition on multiple occasions. Wound infections and mastitis in camels associated with GBS are widespread and should ideally be treated with antimicrobials other than tetracycline in East Africa.     

Development and application of functional markers in maize

Summary Functional markers (FMs) are derived from polymorphic sites within genes causally involved in phenotypic trait variation (Andersen, J.R. & T. Lübberstedt, 2003. Trends Plant Sci 8: 554–560). FM development requires allele sequences of functionally characterized genes from which polymorphic, functional motifs affecting plant phenotype can be identified. In maize and other species with low levels of linkage disequilibrium, association studies have the potential to identify sequence motifs, such as a few nucleotides or insertions/deletions, affecting trait expression. In one of the pioneering studies, nine sequence motifs in the dwarf8 gene of maize were shown to be associated with variation for flowering time (Thornsberry, J.M., M.M. Goodman, J. Doebley, S. Kresovich, D. Nielsen & E.S. Buckler, 2001. Nat Genet 28: 286–289). Proof of sequence motif function can be obtained by comparing isogenic genotypes differing in single sequence motifs. At current, the most appropriate approach for this purpose in crops is targeting induced local lesions in genomes (TILLING) (McCallum, C.M., L. Comai, E.A. Greene & S. Henikoff, 2000. Nat Biotechnol 18: 455–457). In central Europe, maize is mainly grown as forage crop, with forage quality as major trait, which can be determined as proportion of digestible neutral detergent fiber (DNDF). Brown midrib gene knock out mutations have been shown to be beneficial for forage quality but disadvantageous for overall agronomic performance. Two brown midrib genes (bm1 and bm3) have been shown to be involved in monolignol biosynthesis. These two and additional lignin biosynthesis genes have been isolated based on sequence homology. Additional candidate genes putatively affecting forage quality have been identified by expression profiling using, e.g., isogenic bm lines. Furthermore, we identified an association between a polymorphism at the COMT locus and DNDF in a collection of European elite inbred lines.     

Development of new real-time PCR methods for detection of Decapod iridescent virus 1 in shrimp

Novel Decapod Iridescent virus (DIV1) infections emerged in mainland China around 2014 and have devastated shrimp aquaculture operations in Chinese coastal provinces. In 2020, DIV1 has spread to Taiwan with devastating results to shrimp and crayfish farms, in addition to being found in wild caught Penaeus monodon from the Indian Ocean. This trend is a major cause for concern and an urgent reminder to expand the tools needed to monitor the spread of DIV1 globally. Here, we describe a set of four different real-time polymerase chain reaction (PCR) assays positioned across the genome of DIV1 to detect the virus in shrimp tissues. All four assays show a wide dynamic range and high analytical sensitivity and specificity. In addition, the newly developed assays show excellent diagnostic sensitivity and specificity in clinical Litopenaeus vannamei samples of North Asian origin. The new molecular toolset will enhance global capabilities to monitor the spread of DIV1 and ultimately be used as an early warning system for farmers and authorities to engage in appropriate risk mitigation strategies.     

Long-term nitrogen balance for pearl millet (Pennisetum glaucum L.) in an acid sandy soil of Niger

On acid sandy soils of Niger (West Africa) fertilizer N recovery by pearl millet (Pennisetum glaucum L.) is often more than 100 per cent in years with normal or above average rainfall. Biological nitrogen fixation (BNF) by N₂-fixing bacteria may contribute to the N supply in pearl millet cropping systems. For a long-term field experiment comprising treatments with and without mineral fertilizer (F) and with and without crop residue application (CR) a N balance sheet was calculated over a period of six years (1983-1988). After six years of successive millet cropping total N uptake (36-77 kg N ha^?1 yr^?1) was distinctly higher than the amount of fertilizer N applied (30 kg N ha^?1 yr^?1). The atmospheric input of NH₄-N and NO₃-N in the rainwater was about 2 kg N ha_?1 yr^?1, 70 % in the form of NH₄-N. Gaseous NH₃ losses from urea (broadcast, incorporated) were estimated from other experiments to amount to 36 % of the fertilizer N applied. Nitrogen losses by leaching (15 to > 25 kg N ha^?1 yr^?1) were dependent on the treatment and on the quantity and distribution of single rainfall events (>50 mm). Decline in total soil N content (0-60 cm) ranged from 15 to 48 kg N ha^?1 yr^?1. The long-term N balance (1983-1988) indicated an annual net gain between 6 (+CR-F) and 13 (+CR+F) kg N ha^?1 yr^?1. For the control (-CR-F) the long-term N balance was negative (10 kg N ha^?1 yr^?1). In the treatment with crop residues only, the N balance was mainly determined by leaching losses, whereas in treatments with mineral fertilizer application the N balance depended primarily on N removal by the millet crop. The annual net gain in the N balance increased from 7 kg ha^?1 with mineral fertilizer to 13 kg ha^?1 in the combination mineral fertilizer plus crop residues. In both the rhizosphere and the bulk soil (0-15 cm), between 9 and 45% of the total bacterial population were N₂-fixing (diazotrophic) bacteria. The increased N gain upon crop residue application was positively correlated with an increase in the number of diazotrophic and total bacteria. The data on bacterial numbers suggest that the gain of N in the longterm N balance is most likely due to an N input by biological nitrogen fixation. In addition, evidence exists from related studies that the proliferation of diazotrophs and total bacteria in the rhizosphere due to crop residue application stimulated root growth of pearl millet, and thus improved the phosphorus (P) acquisition in the P deficient soil.     

Is N‐feedback involved in the regulation of nitrogenase activity in Medicago truncatula?

To determine whether senescing leaves provoke an active nitrogen (N) remobilization that results in the reduction of nitrogenase activity, 60% of Medicago truncatula lower leaves were either darkened or individually excised for two weeks. Although a considerable amount of N was remobilized, N₂ fixation activity was found to be increased to maintain the N source/sink balance, indicating an absence of the negative N‐feedback regulation of nitrogenase activity in the senescing M. truncatula.     

Effect of mineral nitrogen fertilizer forms on N2O emissions from arable soils in winter wheat production

Nitrogen fertilizers are supposed to be a major source of nitrous oxide (N₂O) emissions from arable soils. The objective of this study was to compare the effect of N forms on N₂O emissions from arable fields cropped with winter wheat (Triticum aestivum L.). In three field trials in North‐West Germany (two trials in 2011/2012, one trial in 2012/2013), direct N₂O emissions during a one‐year measurement period, starting after application of either urea, ammonium sulfate (AS) or calcium ammonium nitrate (CAN), were compared at an application rate of 220 kg N ha^?1. During the growth season (March to August) of winter wheat, N₂O emission rates were significantly higher in all three field experiments and in all treatments receiving N fertilizer than from the non‐fertilized treatments (control). At two of the three sites, cumulative N₂O emissions from N fertilizer decreased in the order of urea > AS > CAN, with emissions ranging from 522–617 g N ha^?1 (0.24–0.28% of applied fertilizer) for urea, 368–554 g N ha^?1 (0.17–0.25%) for AS, and 242–264 g N ha^?1 (0.11–0.12%) for CAN during March to August. These results suggest that mineral nitrogen forms can differ in N₂O emissions during the growth period of winter wheat. Strong variations in the seasonal dynamics of N₂O emissions between sites were observed which could partly be related to weather events (e.g., precipitation). Between harvest and the following spring (post‐harvest period) no significant differences in N₂O emissions between fertilized and non‐fertilized treatments were detected on two of three fields. Only on one site post‐harvest emissions from the AS treatment were significantly higher than all other fertilizer forms as well as compared to the control treatment. The cumulative one‐year emissions varied depending on fertilizer form across the three field sites from 0.05% to 0.51% with one exception at one field site (AS: 0.94%). The calculated overall fertilizer induced emission averaged for the three fields was 0.38% which was only about 1/3 of the IPCC default value of 1.0%.