Diversity of laccase genes from basidiomycetes in a forest soil

Fungal oxidative exo-enzymes lacking substrate specificity play a central role in the cycling of soil organic matter. Due to their broad ecological impact and available knowledge of their gene structure, laccases appeared to be appropriate markers to monitor fungi with this kind of oxidative potential in soils. A degenerate PCR-primer pair Cu1F/Cu2R, specific for basidiomycetes, was designed to assess directly the diversity of laccase genes in soils. PCR amplification of mycelial cultures and fruit-bodies of a wide spectrum of basidiomycetes, covering all functional groups (saprophytes, symbionts, and pathogens), produced multiple DNA fragments around 200 bp. A neighbor-joining tree analysis of the PCR-amplified laccase sequences showed a clear species-specificity, but also revealed that most fungal taxa possess several laccase genes showing a large sequence divergence. This sequence diversity precluded the systematic attribution of amplified laccase of unknown origin to specific taxa. Amplification of laccase sequences from DNA, extracted from a brown (moder) forest soil, showed a specific distribution of laccase genes and of the corresponding fungal species in the various soil horizons (O_h, A_h, B_v). The most organic O_h-horizon displayed the highest gene diversity. Saprophytic fungi appeared to be less widespread through the soil horizons and displayed a higher diversity of laccase genes than the mycorrhizal ones.     

Phosphorus availability,root exudates,and microbial activity in the rhizosphere

Field and pot experiments showed that the P demand of wheat is highest in early stages of growth (up to 1.67 μg P per cm² root surface and day). The needed orthophosphate ions H₂PO₄? and HPO₄²-move from soil to the root by diffusion. This process is controlled by the concentration gradient of the diffusible phosphate and the effective diffusion coefficient according to Pick's first law. Root excretions (rhizodeposition) are able to affect both characteristics. The water soluble portion of rhizodeposition contains more than 50% of up to 8 different sugars, 10–40% carboxylic acids and 10–15 amino acids and amides. The composition varies in dependence on the age of the root parts and on nutrition (Zea mays L., Brassica napus L., Pisum sativum L.). Diffusion experiments using small soil blocks showed that 50–75% of the root exudates were decomposed by respiration within 3 days. The rest was largely chemically converted. Originally present sugars disappeared. Due to the biosynthesis of different organic acids from the individual sugars the mobilisation of Ca₃(PO₄)₂ by Pantoea agglomerans increased when the sugar mixture was derived from the rhizodeposition of P deficient plants with more pentoses instead of glucose and fructose (mainly effect of anions). In the rhizosphere therefore a mixture of rhizodeposition and its conversion products exists which affects the binding of phosphorus in soil and the P transport to the root. This should be considered both for the development of new soil extractants and for modelling the P supply to plants.     

Formation of early and advanced Maillard reaction products correlates to the ripening of cheese

The present study deals with the characterization of the ripening of cheese. A traditional German acid curd cheese was ripened under defined conditions at elevated temperature, and protein and amino acid modifications were investigated. Degree of proteolysis and analysis of early [Amadori compound furosine (6)] and advanced [N(ε)-carboxymethyllysine (4), N(ε)-carboxyethyllysine (5)] Maillard reaction products confirmed the maturation to proceed from the rind to the core of the cheese. Whereas 6 was decreased, 4 and 5 increased over time. Deeper insight into the Maillard reaction during the ripening of cheese was achieved by the determination of selected α-dicarbonyl compounds. Especially methylglyoxal (2) showed a characteristic behavior during storage of the acid curd cheese. Decrease of this reactive structure was directly correlated to the formation of 5. To extend the results of experimental ripening to commercial cheeses, different aged Gouda types were investigated. Maturation times of the samples ranged from 6 to 8 weeks (young) to more than 1 year (aged). Again, increase of 5 and decrease of 2 were able to describe the ripening of this rennet coagulated cheese. Therefore, both chemical parameters are potent markers to characterize the degree of maturation, independent of coagulation.     

Constitutive emission of the aphid alarm pheromone, (E)-β-farnesene, from plants does not serve as a direct defense against aphids

Background  

The sesquiterpene, (E)-β-farnesene (EBF), is the principal component of the alarm pheromone of many aphid species. Released when aphids are attacked by enemies, EBF leads aphids to undertake predator avoidance behaviors and to produce more winged offspring that can leave the plant. Many plants also release EBF as a volatile, and so it has been proposed that this compound could act to defend plants against aphid infestation by 1) deterring aphids from settling, 2) reducing aphid performance due to frequent interruption of feeding and 3) inducing the production of more winged offspring. Here we tested the costs and benefits of EBF as a defense against the green peach aphid, Myzus persicae, using transgenic Arabidopsis thaliana lines engineered to continuously emit EBF.     

Comparison of different extractants for the determination of inorganic sulphate in gypsum‐free agricultural soils

Four frequently used extractants (H₂O, 0.1 M NaCl, 0.016 M KH₂PO₄, and 0.5 M NaHCO₃) as well as different extraction conditions have been tested for sulphate extraction from gypsum‐free agricultural soils. Water is the preferable extractant for soils with pH > 6. Two extraction steps have to be carried out for complete extraction (> 95%). A 0.016 M KH₂PO₄ solution was found to be the most efficient extractant for soils with a pH < 6 within a single extraction step. A shaking frequency of 170 min^‐1 and a duration of extraction of 4 hours are the optimized conditions for the sulphate extraction with H₂O and KH₂PO₄ solution.     

Combination of TLC and HPLC-MS/MS methods. Approach to a rational quality control of Chinese star anise

In this study, a methodological approach for an effective and reliable quality control of Chinese star anise (Illicium verum Hook. F.) is developed and validated. A combined method of TLC and HPLC-MS/MS was used for differentiation of various Illicium species, especially Chinese and Japanese star anise. Species can be distinguished by their TLC flavonoid pattern. A sensitive and selective HPLC/ESI-MS/MS method was developed for the detection and quantification of lower admixtures of I. anisatum and of further toxic Illicium species at a low concentration range using the sesquiterpene lactone anisatin as a marker. The proposed assay includes a solid-phase extraction cleanup procedure with a high recovery (>90%). Chromatographic separation of anisatin was carried out on a C18 column, followed by MS detection using ESI in negative mode. The precursor/product ion transitions m/z 327 --> 127 (quantifier) and m/z 327 --> 297 (qualifier) were monitored. Statistical evaluation of this multireaction-monitoring procedure reveals good linearity and intra- and interday precision. The limits of detection and quantification are 1.2 and 3.9 microg/kg, respectively.     

Detection of donor effects in a rye introgression population with genome‐wide prediction

Introgression populations are developed to make genetic resources for breeding purposes available. In the case that the number of donor segments exceeds the number of lines, genome‐wide prediction (GWP) methods are suggested as promising for the analysis of such populations. Our objectives were to characterize a rye introgression population with the Rye5K SNP assay and to apply a GWP model with a modification of the restricted maximum likelihood procedure that yields heteroscedastic variances to detect significant donor effects. The introgression lines (ILs) carried on average 4.6 donor segments with a mean length of 27 cM and represented 94% of the donor genome. Two donor effects were detected that significantly increased thousand‐kernel weight. We found four donor effects for protein, total pentosan and starch content that can improve baking quality. Three donor effects for protein content were observed for improving feeding purposes and one donor effect for starch content to improve ethanol production. The effects were localized to small genomic regions. Consequently, these ILs can improve rye breeding by directly employing them in breeding programmes for variety development.