Comparative effect of low temperature on virulence and twitching motility of Ralstonia solanacearum strains present in Florida

Ralstonia solanacearum causes bacterial wilt on a wide range of plant hosts. Most strains of R. solanacearum are nonpathogenic below 20°C; however, Race 3 Biovar 2 (R3B2) strains are classified as quarantine pathogens because of their ability to infect crops, cause disease, and survive in temperate climates. We have identified race 1 biovar 1 Phylotype IIB Sequevar 4 strains present in Florida which were able to infect and produce wilt symptoms on potato and tomato at 18°C. Moreover they infected tomato plants at rates similar to strains belonging to R3B2. We determined that strains naturally nonpathogenic at 18°C were able to multiply, move in planta, and cause partial wilt when inoculated directly into the stem, suggesting that low temperature affects virulence of strains differently at early stages of infection. Bacterial growth in vitro was delayed at low temperatures, however it was not attenuated. Twitching motility observed on growing colonies was attenuated in nonpathogenic strains at 18°C, while not affected in the cool virulent ones. Using pilQ as a marker to evaluate the relative expression of the twitching activity of R. solanacearum strains, we confirmed that cool virulent strains maintained a similar level of pilQ expression at both temperatures, while in nonpathogenic strains pilQ was downregulated at 18°C.     

Quantitative assessment of hydrolase production and persistence in soil

The aim of this work was to calculate indices of hydrolase production (Pr) and persistence (Pe) through simple arithmetical calculations. Changes in acid and alkaline phosphomonoesterase, phosphodiesterase, urease, protease, and β-glucosidase activities were monitored under controlled conditions in seven soils with a wide range of properties, in which microbial growth was stimulated by adding glucose and nitrogen. Glucose mineralization was monitored by CO₂–C evolution, and microbial growth was quantified by determining the soil adenosine triphosphate (ATP) content. Hydrolase Pr and Pe indices were numerically quantified by the following relationships: Pr = H / t _H and Pe = (r / H)Δt, respectively, where H indicates the peak value of each measured hydrolase activity, t _H is the time of the peak value, r indicates the residual activity value, and Δt is the time interval t _r − t _H, where t _r is the time of the residual activity value. Addition of glucose and N-stimulated soil respiration increased ATP content and stimulated the production of the measured hydrolase activities in all soils; the measured variable reached a maximum value and then decreased, returning to the value of the control soil. Apart from β-glucosidase activity, whose activity was not stimulated by glucose and N addition, the other measured hydrolase activities showed a trend that allowed us to calculate the Pr and Pe indices using the above-mentioned equations. Acid phosphomonoesterase and protease Pr values were significantly higher in soils under forest or set aside management; the alkaline phosphomonoesterase and phosphodiesterase Pr values were generally higher in the neutral and alkaline soils, and the urease Pr values showed no obvious relationships with soil pH or management. Concerning the persistence of enzyme activities, Pe values of the acid phosphomonoesterase activity were significantly higher in the acidic soils, and those of urease activity were higher in acidic soils and the Bordeaux neutral soil. No relationships were observed between Pe values of alkaline phosphomonoesterase, phosphodiesterase, or protease activities and soil pH or management. The different responses of hydrolases were discussed in relation to soil properties, microbial growth, and regulation at the enzyme molecular level.     

Application of a new sediment contact test with <Emphasis Type="Italic">Myriophyllum aquaticum</Emphasis> and of the aquatic <Emphasis Type="Italic">Lemna</Emphasis> test to assess the sediment quality of Lake Skadar

Goal, Scope and Background  Situated in the transboundary belt between Montenegro and Albania, Lake Skadar is the largest freshwater reservoir in Southeastern Europe. Because of the wide range of endemic, rare or endangered plant and animal species it supports, Lake Skadar and its extensive adjacent wetlands are internationally recognised as a site of significance and importance (Ramsar site). Within the last 10 to 20 years, Lake Skadar was exposed to intensive pollution. For the assessment of the ecotoxic load of the sediments sampled in Lake Skadar, a triad approach was recently applied. Overall, a complex spectrum of ecotoxic loads was elucidated. The aim of the present study was to use plant-based bioassays for assessing the sediment quality of Lake Skadar in order to facilitate and complement the triad test battery. The newly developed sediment contact test with Myriophyllum aquaticum and the aquatic growth inhibition test with Lemna minor were applied to native sediments and pore water, respectively, allowing the investigation of different toxicity-effects caused by particle-bound pollutants as well as pollutants in the interstitial water. This investigation is the first application of the novel sediment contact test with Myriophyllum aquaticum to lake sediments. Methods  Sediment samples were taken from nine selected sites at Lake Skadar and investigated by the sediment contact assay with Myriophyllum aquaticum. The pore water was extracted from these sediment samples to be analysed in the aquatic growth inhibition test with Lemna minor. The results of the sediment contact tests were compared with each other and with those of the aquatic growth inhibition test. Results and Discussion  Both applied macrophyte biotests revealed distinct changes in the growth behaviour of the two macrophytes subsequent to the exposure to the investigated natural sediments of Lake Skadar. The Myriophyllum sediment contact test revealed significant toxicity in the sediment samples from Radus and Kamenik, whereas the aquatic Lemna test showed inhibition effects for the samples from Sterbeq, Plavnica and Kamice. Data obtained with the newly developed Danio rerio contact test and the Arthrobacter globiformis contact test confirmed the Myriophyllum results. Analyses of the heavy metal content in the sediments revealed low or moderate contamination levels. Correlation analyses between the content of heavy metals in the sediments and growth inhibition of Myriophyllum aquaticum showed a significant correlation between Cr concentrations and growth inhibition. Comparable findings are available for a German river system. In contrast, no significant correlation between inhibition rates and concentration of metals could be observed with Lemna minor. Conclusions  It was shown that the newly developed sediment contact test with Myriophyllum aquaticum is applicable to lake sediments. In both the sediment contact test with Myriophyllum aquaticum on whole sediments and the aquatic growth inhibition test with Lemna minor on pore water, plant growth was influenced by the natural sediments and its components. Therefore, both test systems were found to be suitable for the detection of phytotoxic effects upon exposure to sediments. Myriophyllum aquaticum as test organism of the contact test grows directly in the sediment without an additional water-layer. Thus, it is able to detect toxicity caused by particle-bound phytotoxic substances as well as pore water-related contamination, while the floating Lemna minor can only detect effects emanating from pore water. Significant differences of the results were observed between these two test systems and, accordingly, the two different exposure scenarios. Hence, none of the tests can replace the other one and, as a consequence, both should be included into a test battery for the assessment of sediment toxicity. Recommendations and Perspectives  Both plant assays were shown to be reliable tools for the evaluation of the eco-toxicological risk potentials of pore water and solid-phase sediment. They should become a complement to the standardised test battery generally used for comprehensive hazard assessment. ESS-Submission Editor: Dr. Ulrike Kammann (ulrike.kammann@ifo.bfa-fisch.de)     

Tradeoffs in the Rehabilitation of a Succulent Karoo Rangeland

Rangeland rehabilitation has multiple, sometimes conflicting goals, such as the reestablishment of the predisturbance vegetation, soil protection, and forage production. The rehabilitation techniques should be also cost‐effective and practicable. Given the difficulties and high costs of restoring Succulent Karoo rangelands and the continuously high grazing pressure in the communal lands, tradeoffs should be accepted in the achievement of these goals. We tested the capability of paddock manure redistribution to reverse degradation trends in a heavily grazed Succulent Karoo rangeland in South Africa. Over 3 years, the effects of the manure application were compared with areas planted with mature shrubs as a benchmark for a predisturbance vegetation structure and with four popular rehabilitation techniques: (1) livestock exclusion; (2) brushpacking (coverage of dead shrubs); (3) mineral fertilizing; and (4) microcatchment construction. Manure was, besides planting, the only treatment that resulted in a significant increase in drought‐resistant vegetation cover, but it compromised the dominance of native vegetation. In the manure plots, a pasture‐like vegetation of non‐native forage plants (which germinated mainly from seeds in the dung), developed (foremost Atriplex semibaccata). Manure application counteracted erosion as effectively as the planted shrubs and brushpacks. Expected negative side effects such as a decrease in plant species richness or salinization of topsoil were not detected. We also checked the potential of topsoil salinization by the halophytic A. semibaccata and found it to be low. For sites where a decrease in grazing pressure is unrealistic under current land tenure, redistribution of manure should be further explored to mitigate acute symptoms of degradation. Copyright © 2013 John Wiley & Sons, Ltd.     

Molekulare Diagnostik in der Lebensmittel��berwachung

Molecular diagnostic is frequently a confirmed part in the food microbiological survey of the official food safety control. So the diagnostic tools are used for a fast screening for presence or absence of pathogens or to differentiate the isolates.     

Einfluss von mechanischen Ber��hrungsreizen auf das Wachstum von K��chenkr?utern im Topf

For a compact growth of many potted plants you often have to use chemical growth inhibitors. Various species of herbs tend to an undesirable extension growth too. In general, the use of chemical inhibitors is prohibited in the production of potted herbs. For several years at the State Horticultural College and Research Institute Heidelberg there is employed an alternative method of mechanical stimulation treatment for a compact growth of potted plants. The greatest reduction of elongation was obtained, when the plants were treated with breaks in the early morning. In 2009 and 2010, the employees of the trial station proofed the effect of mechanical stimulation (Thigmomorfogenesis) to a large assortment of organically produced potted herbs. Green and red varieties of basil, parsley, coriander, chervil, lemon balm (2009) and four different varieties of mint (2010) were stimulated with a machine one week after beginning culture continuously 108 times per day. Compared to the untreated plants all mechanically stimulated herb species and varieties showed a tendency or significantly reduced elongation growth and therefore more stability. Depending on species and varieties the quality of these plants was visibly improved.     

Prokaryotic chromosomes and disease

Recent insights into bacterial genome organization and function have improved our understanding of the nature of pathogenic bacteria and their ability to cause disease. It is becoming increasingly clear that the bacterial chromosome constantly undergoes structural changes due to gene acquisition and loss, recombination, and mutational events that have an impact on the pathogenic potential of the bacterium. Even though the bacterial genome includes additional genetic elements, the chromosome represents the most important entity in this context. Here, we will show that various processes of genomic instability have an influence on the many manifestations of infectious disease.     

Phosphate aquisition by red clover and black mustard on a humic podzol

Recent investigations have shown that phosphate (P) mobilization by root exudates is an important feature of genotypes to acquire P even in soils of low‐P availability. We, therefore, investigated P mobilization processes in the rhizosphere of red clover (Trifolium pratense L.) and black mustard (Brassica nigra L.) on a humic podzol. As measured by the Kuchenbuch‐Jungk method (Kuchenbuch and Jungk, 1982), both species accumulated similar quantities of citrate (12 μmol/g soil) in the rhizosphere in about 1 mm distance from the soil‐root interface. Despite of similar concentrations of P‐mobilizing citrate in the rhizosphere of both species, red clover took up nearly the two‐fold of P compared to black mustard. Differences in rhizosphere pH were determined between both species. Black mustard did not acidify the rhizosphere, whereas red clover decreased the pH in the rhizosphere from 5.8 to about 4.0 (in 0.01M CaCl₂). The simultaneous acidification and excretion of citrate compared to citrate excretion alone had consequences for P mobilization processes in the rhizosphere. Phosphate mobilization from the soil solid phase was higher at higher pH. Thus, the citrate‐induced P desorption was not the limiting step in P acquisition by red clover and black mustard. Calculations of P distribution in the soil solution between free ortho‐P and humic‐associated P showed that at higher pH most of the P was associated with dissolved humic substances, whereas at pH < 5, most of the P was present as free ortho‐P. These P species can readily be taken up by the roots whereas humic‐associated P must probably be desorbed from the humic surface before uptake. Phosphate species calculations, therefore, explained the higher P uptake of red clover compared to black mustard. Aluminum species distribution calculations in the soil solution further show that even at pH < 5.0 in the soil solution, citrate strongly complex Al and thereby reduce the activity of monomeric Al species. The excretion of citrate can, therefore, counteract the root induced acidification of the rhizosphere with respect to Al toxicity.