Competence of <Emphasis Type="Italic">Xanthomonas campestris</Emphasis> from Cruciferous Weeds and Wallflower (<Emphasis Type="Italic">Erysimum cheiri</Emphasis>) to Induce Black Rot in Cabbage

Aphanomyces euteiches Drechsler is an oomycete pathogen of leguminous crops that causes root rot, a severe disease of pea (Pisum sativum L.) worldwide. An improved understanding of the genetic structure of A. euteiches populations would increase knowledge of pathogen evolution and assist in the design of strategies to develop pea cultivars and germplasm with stable disease resistance. Twenty six primers pairs were used to amplify Sequence Related Amplified Polymorphisms (SRAP) among 49 A. euteiches isolates sampled from pea. A total of 190 polymorphic SRAP bands were generated, of which 82 were polymorphic between all the A. euteiches isolates. The percentage of polymorphic bands per primer pair ranged from 22 to 75%. According to the PIC value estimated for each marker, 60% of the SRAP markers were highly to reasonably informative (PIC > 0.25). Genetic structure of A. euteiches populations sampled in different American and French locations showed low to high genetic diversity within populations. The largest variation occurred within countries, with a total estimated genetic diversity of 0.477 and 0.172 for American and French populations, respectively. This was particularly evident from a principal component analysis (PCA) and a Minimum Spanning Networks (MSN) based on genetic profiles of isolates, which generated two different clusters, one corresponding to the French isolates and four American isolates (MV1, MV5, MV7, Ath3), and the other to American isolates. A. euteiches populations from cultivated pea in France appeared as a single unstructured population, whereas American isolates of A. euteiches diverged into three different populations.     

Correlating phosphorus extracted by simple soil extraction methods with foliar phosphorus concentrations of Picea abies (L.) H. Karst. and Fagus sylvatica (L.)

Phosphorus (P) concentrations in needles and leaves of forest trees are declining in the last years in Europe. For a sustainable forest management the knowledge of site specific P nutrition/availability in forest soils is vital, but we are lacking verified simple methods for the estimation of plant available P. Within this study, four soil P extraction methods [water ( ), double‐lactate (P_lac), citric acid (P_cit), and sodium bicarbonate ( )], as well as total P content of the soil (P_tot) were tested to investigate which method is best correlated with foliar P concentrations of spruce [Picea abies (L.) H. Karst.] and beech [Fagus sylvatica (L.)]. Mineral soil samples from 5 depth levels of 48 forest sites of the Bavarian sample set of the second National Forest Soil Inventory (BZE II) were stratified according to tree species (spruce and beech) and soil pH (pH < 6.2 and > 6.2), covering the whole range of P nutrition. The extractable amount of P per mass unit of soil increased in the order << P_lac < < P_cit, decreased with soil depth, and was higher in soils with pH < 6.2. Citric acid extracted up to 10% of P_tot in acidic soils. Whereas P_cit delivers adequate regression models for P nutrition in the case of spruce (R² up to 0.53) and beech (R² up to 0.58) for acidic soils, shows good results for spruce growing on acidic soils (R² up to 0.66) and for beech on soils with pH > 6.2 (R² up to 0.57). P_lac produces adequate models only for beech on high pH soils (R² up to 0.64), while did not produce acceptable regression models. P_tot seems suitable to explain the P nutrition status of beech on acidic (R² up to 0.62) and alkaline soils (R² up to 0.61). Highest R²s are obtained mostly in soil depths down to 40 cm. As and P_cit showed good results for both investigated tree species, they should be considered preferentially in future studies.     

Changes in forest soil organic matter quality affected by windstorm and wildfire

Purpose

Windthrows and fires are major natural disturbances in forest ecosystems, which can affect organic matter in the surface and the mineral layer of forest soil. The main aim of this study was to evaluate the changes occurring in the structure and properties of humic acid (HA) in the lands where windthrows and wildfires occurred.

Materials and methods

In November 2004, the forest in the area of 12,000 ha in the Tatra National Park, Slovakia, was seriously damaged by northern wind gusts exceeding 200 km/h. In July 2005, a wildfire broke out in a 220 ha of wind-damaged area. The HAs have been isolated from four research plots: (a) the area where the fallen trees were removed (EXT); (b) an area after windstorm covered by wood from struck trees (NEX), left for spontaneous succession; (c) an area after extracted timber, damaged by the surface wildfire (FIR); and (d) a reference intact spruce forest area (REF). Changes in the chemical structure of the HAs isolated from the research plots were determined on the basis of elemental analysis and UV-Vis, EPR, IR, and ¹³C NMR spectroscopy.

Results and discussion

All used analytical methods showed a decrease in the humification degree of the humic acids extracted from the soils where the spruce forest has been affected by a wildfire and a windthrow. In the case of the control sample HA (REF), the calculated atomic H/C and O/C ratios and the degree of aromaticity (α) calculated from the ¹³C NMR spectra were higher, indicating higher aromaticity of HA from the REF area. The more complex and developed structure of REF HA was confirmed by the higher value of E^1%₆ and the lower E4/E6 ratio obtained using UV-Vis spectroscopy. Also, the higher g-parameters determined from the EPR spectra of the stable radicals present in HA confirmed the lower aromaticity on the plots that have been subjected to the calamities. The ¹³C NMR spectra and the elemental analysis show that the structure of the HA extracted from the NEX plot is the closest to the REF.

Conclusions

The results of the systematic research showed significant changes in the structure of HA taken from spruce forest soils that were subjected to windstorms and fires. An enrichment of the HAs in aliphatic carbon and so a lower humification degree of the organic matter in the areas calamity-affected were observed. The results clearly indicate that the HAs extracted from the disturbed plots of the spruce forest are not as stable as those extracted taken from the control plot.

     

Importance of soil extractable phosphorus distribution for mature Norway spruce nutrition and productivity

Phosphorus is an essential nutrient for forest growth. In this study, we assessed the impact of soil extractable phosphorus using two simple extraction methods on nutrition and productivity of Norway spruce in sixteen mature forest stands on different bedrocks and soils in Bavaria, Southern Germany. Representative trees were sampled for needles, twigs, branches, stem bark, and stem wood. Total phosphorus content in the tree parts and soil phosphorus stock extractable with citric acid and sodium bicarbonate up to a soil depth of 80 cm were determined. We found that easily soil extractable phosphorus is a suitable indicator for estimating phosphorus uptake and stand productivity in Norway spruce. In contrast, organic layer phosphorus showed no significant correlation with aboveground biomass phosphorus contents. In the biomass, the highest phosphorus contents were measured in young needles and twigs, but the highest correlation with soil phosphorus was detected for phosphorus contents in needles and bark. The stock of phosphorus extracted by citric acid down to 40 cm soil depth revealed the best correlation with phosphorus in needles and bark. Therefore, as a supplemental or alternative method to needle analysis, our study suggests the use of phosphorus contents in stem bark to evaluate tree phosphorus nutrition. These results highlight the suitability of the citric acid soil extraction method to characterize plant available phosphorus in Norway spruce ecosystems.     

Suitability of two agglomerated commercial microdiets for rearing larvae of different strains of Coregonus lavaretus under cold‐water conditions

The suitability of two agglomerated commercial microdiets (“Gemma micro^®” and “O.range^®” from now on called diet A and diet B) as a starter feed was tested on larvae of European whitefish (Coregonus lavaretus). Two strains (Baltic and Alpine) were tested in a 60‐day experiment under ambient cold‐water conditions (average 7.6 ± 1.9°C). The experiment was carried out in 400‐L PVC tanks with a stocking density of 5,000 larvae per tank. Samples were removed weekly to determine growth, survival and feed acceptance, in addition to water quality. Striking differences between the feeds and different strains were observed. For one diet, growth and survival were distinctively higher in strain 1 compared to strain 2, where it was invariably low with both diets. For the second diet, growth and survival in strain 1 were also low and similar to strain 2. Feed acceptance in strain 1 was found higher compared to strain 2. Final weights were significantly different with strain 1 being heavier; however, in terms of relative growth rate, no differences between strains were found. Strong differences in the performance in terms of growth, survival and feed acceptance of different strains of European whitefish became apparent. Furthermore, the rearing of larvae exclusively on standard agglomerated commercial microdiets under cold‐water conditions is possible, but not every microdiet is suitable.     

Molecular tracing confirms that infection with infectious pancreatic necrosis virus follows the smolt from hatchery to grow‐out farm

Infectious pancreatic necrosis (IPN) is an important restraint to production of salmonids in aquaculture globally. In order to implement efficacious mitigation strategies for control of this disease, it is important to understand infection routes under current production systems. IPN virus has been shown to be transmitted vertically in Rainbow trout, from broodstock to fingerlings in hatcheries, and there is circumstantial evidence suggesting that vertical transmission can also occur in Atlantic salmon, in addition to horizontal transmission between grow‐out fish in farms. In this study, we show that the smolt carries infection with IPN from hatchery to the marine farm. We do this by comparing sequences from fish groups taken both in hatcheries and on corresponding marine grow‐out farms. We use statistical analysis to prove that sequences obtained from the same fish group in both hatchery and marine farm are more similar than sequences obtained from random fish groups on hatcheries and marine farms.     

Immunogene expression in head kidney and spleen of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) following thermal stress and challenge with Gram-negative bacterium, <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

To evaluate the effect of thermal and microbial stress on the immune response of common carp (Cyprinus carpio L.), relative mRNA expression level of pro-inflammatory cytokines [tumor necrosis factor alpha (TNF-α) and interleukin (IL)-1β] and other genes related to immune or stress response [inducible nitric oxide synthase (iNOS), heat shock protein 70 (Hsp70), superoxide dismutase one (SOD1), and glucocorticoid receptor (GR)] was measured by quantitative PCR (qPCR). In addition, total protein and total immunoglobulin level in blood plasma of experimental common carp was also assayed. All the above parameters were estimated 24 h post-challenge with Gram-negative bacterium, Aeromonas hydrophila. Common carp (54.89?±?6.90 g) were initially exposed to 20 °C (control group) and 30 °C (thermal stress group) water temperature for 30 days, followed by experimental challenge with 2.29?×?10⁸ colony forming unit/mL (CFU/mL; LD₅₀ dose) of A. hydrophila. Exposure of fish to thermal stress and subsequently challenge with A. hydrophila significantly (P?<?0.05) increases the IL-1β mRNA expression in head kidney and spleen of common carp by ~?39.94 and ~?4.11-fold, respectively. However, TNF-α mRNA expression in spleen decreased ~?5.63-fold in control fish challenged with A. hydrophila. Thermal stress and challenge with bacterium suppresses the iNOS and GR mRNA expression in spleen of common carp. Moreover, significant (P?<?0.05) increase in total protein content of blood plasma (~?43 mg/g) was evident in fish exposed to thermal stress and challenged with A. hydrophila. In conclusion, our study highlights the importance of elevated temperature stress and microbial infection in differential regulation of expression of several immunogenes in common carp.     

Ripening reduces the shrinkage of processed dredged material

Purpose

The utilization of dredged material in dike construction as a substitute for traditionally used materials is considered as an option to preserve natural resources such as marsh sediments. As a prerequisite for this application, the equivalency with respect to soil physical and mechanical properties of the materials must be assessed. Previous investigations have shown pronounced differences in shrinkage behavior and desiccation cracking between sediments and dredged material. The key objective of the study was to assess whether shrinkage of processed dredged material can be reduced by further processing, i.e., dewatering, which can be referred to as ripening.

Materials and methods

To compare the shrinkage behavior of the materials, three different methods of different scales were applied. Small-scale methods conducted were the standard procedure for the determination of the shrinkage limit and the determination of the coefficient of linear extensibility (COLE_rod). Large-scale shrink-swell experiments were carried out in a specially constructed test system with 90 l capacity for a period of up to 385 days. Here the materials were ripened, i.e., air-dried, until shrinkage almost ceased, and a rewetting-air-drying cycle was conducted. Shrinkage and swelling were determined during the processes by measuring the changes in volume. On the ripened materials, COLE_rod was determined.

Results and discussion

The experiments show that the shrinkage behavior of processed dredged material can be ameliorated by ripening. COLE_rod of the ripened materials were about 20–80% lower than COLE_rod of the un-ripened materials. The large-scale shrink-swell experiments showed that shrinkage in the second drying cycle amounted to less volume than in the first drying cycle and that shrinkage behavior in contrast to the first drying cycle, where pronounced proportional shrinkage was observed, was dominated by structural and residual shrinkage in this cycle.

Conclusions

Ripening of processed dredged material is considered a useful pre-treatment option to ameliorate the shrink-swell behavior of processed dredged material and to obtain a better functional equivalency with traditionally used dike construction materials such as fine-grained marsh sediments.

     

Influence of temperature,humidity duration and growth stage on the infection and mycotoxin production by Fusarium langsethiae and Fusarium poae in oats

High occurrence of Fusarium poae (FP) and Fusarium langsethiae (FL) and their mycotoxins nivalenol (NIV) and T-2/HT-2 have been observed in Swiss oats. Early prediction of mycotoxin levels is important for farmers and the cereal industry to minimize the risk of contaminated food and feed. Therefore, climate chamber experiments were conducted to investigate the influence of different temperatures (10, 15, 20 °C) and durations (4, 8, 12 h) at 99% relative humidity (RH) on the infection of oats with FP and FL. In addition, to discover the most susceptible period of oats, artificial FL inoculations were conducted at different growth stages. Field experiments were performed to observe the dispersal of these fungal species within the field and to investigate the weather conditions that influence the dispersal. The climate chamber experiments revealed higher contamination with NIV and T-2/HT-2 in the 10 °C treatments and with a prolonged humidity duration of 12 h 99% RH. Inoculations of oat plants at early (DC 61) and mid (DC 65) anthesis, led to higher FL infection and T-2/HT-2 accumulation in the grains compared with treatments at earlier growth stages, which might be due to an increased susceptibility during anthesis. No indication for spore dispersal was observed in the field experiments. The results obtained, together with the cropping factors that influence infection and mycotoxin production, could be used as a first step in developing forecasting models to predict the contamination of oats with the mycotoxins NIV and T-2/HT-2.     

Evaluation of 28 balm and lemon balm (<Emphasis Type="Italic">Melissa officinalis</Emphasis>) accessions for content and composition of essential oil and content of rosmarinic acid

Lemon balm (Melissa officinalis L.) is a well-known aromatic and medicinal plant. Because of its proven sedative, spasmolytic and antiviral effects, it is often used in watery or alcoholic extracts for self-medication or pharmaceutical purposes. Its therapeutic effect is due to the content of essential oil and rosmarinic acid. A set of 28 balm accessions was evaluated for the variability of essential oil content and composition and the content of rosmarinic acid. For analysis of secondary metabolites distillation, gas chromatography and high performance liquid chromatography was used. The content of essential oil in the first cut of the years 2009 and 2010 varied in this study in a range between 0.02 and 0.48% and in the second cut between 0.01 and 0.72%. The rosmarinic acid content of the collection ranged in the year 2009 for the first cut from 3.99 to 8.78% and in the second cut from 2.45 to 6.78%. Ploidy was determined for all accessions and two cytotypes were found: diploid 2n = 2x = 32 (15 accessions), tetraploid 2n = 4x = 64 (13 accessions). Via statistical analyses two chemotypes of essential oil were found: chemotype citral and chemotype germacrene D.