Development of a complete set of disomic rape-radish chromosome-addition lines

After crosses of multiple rape-radish chromosome additions with rape, nine different monosomic additions (2n = 4x = 38 + 1) of individual radish chromosomes, a–i, were isolated, having the genomic background of winter oil seed rape and radish cytoplasm. Extra chromosomes were identified with RAPD markers, specific for individual radish chromosomes. All radish-plasmic monosomic additions, except that of chromosome f, had low seed production due to pistilloid stamens. Rape cytoplasm was substituted by crossing the monosomic additions as pollinators to rape followed by reselection of the radish chromosome additions. The monosomic additions in rape cytoplasm showed normal male fertility. Average transmission rates of radish chromosomes via egg cells and pollen cells were 37% and 27%, respectively. Lowest male transmission was found for chromosomes e and f with 0.04 and 0.01. Monosomic additions were self-pollinated to produce disomic additions. Monosomic and disomic addition progenies were discriminated by a prescreening with quantitative double primer (dp) RAPDs and cytological confirmation of preselected candidates by FISH analysis using a Raphanus-specific probe. Usability of this complete set of nine disomic rape-radish additions (2n = 4x = 38 + 2) was discussed.     

Development of phytoplankton communities and common off-flavors in a biofloc technology system used for the culture of channel catfish (Ictalurus punctatus)

The use of biofloc technology production systems continues to increase in the aquaculture industry worldwide. Recent research demonstrated that outdoor biofloc systems can be used to produce high yields of channel catfish (Ictalurus punctatus). However, studies have not yet been performed to determine the development and composition of phytoplankton communities and related off-flavor problems in these biofloc production systems. In this study, water samples were collected biweekly from May to November and channel catfish samples were collected during harvest in November from nine 18.6 m² biofloc culture tanks. Water and fillet samples were analyzed for levels of the common off-flavor compounds geosmin and 2-methylisoborneol (MIB). The development and composition of phytoplankton communities in each culture tank was also monitored. In addition, water and biofloc samples were evaluated to assess the microbial sources of geosmin and MIB within the culture tanks. Phytoplankton (including algae and cyanobacteria attached to bioflocs) biomass, as determined by concentrations of chlorophyll a in the water, gradually increased in all tanks over time. Phytoplankton communities that developed in the culture tanks were dominated by fast-growing, unicellular and small colonial types of green algae (chlorophytes) and diatoms (bacillariophytes) and slower growing, small colonial types of cyanobacteria (cyanophytes). A positive correlation (p < 0.05) between cumulative feed addition and chlorophyll a concentration was found. Although geosmin and MIB were present in the culture water of each tank during most of the study, levels were typically low and only one tank yielded catfish with geosmin and MIB in their flesh at levels high enough to be designated as off-flavor. A positive correlation (p < 0.05) between cumulative feed addition and MIB concentrations in the water of culture tanks indicates a greater potential for MIB-related off-flavor problems at high feed application rates. The microbial sources responsible for production of geosmin and MIB in the culture tanks remain unknown.     

Triazole fungicides and the selection of resistance to medical triazoles in the opportunistic mould Aspergillus fumigatus

Azole resistance is an emerging problem in the opportunistic mould Aspergillus fumigatus. The triazoles are the most important agents for the management of Aspergillus diseases in humans. Selection for acquired resistance may occur in the hospital setting through exposure to high doses of azoles during azole therapy, but evidence is accumulating that A. fumigatus may become resistant to medical triazoles through environmental exposure to fungicides. The recovery of A. fumigatus isolates resistant to the medical triazoles from azole‐naive patients as well as from the environment strongly indicates an environmental route of resistance selection. Molecule alignment studies have identified five fungicides that share a very similar molecule structure with the medical triazoles, and thus may have selected for mechanisms that confer resistance to both groups of compounds. It is important to explore further the presumed fungicide‐driven route of resistance selection in order to implement effective preventive measures as the prevalence of azole resistance in A. fumigatus continues to increase and causes major challenges in the management of azole‐resistant Aspergillus diseases. Copyright © 2012 Society of Chemical Industry     

Construction of a chromosome-assigned,sequence-tagged linkage map for the radish,Raphanus sativus L. and QTL analysis of morphological traits

The radish displays great morphological variation but the genetic factors underlying this variability are mostly unknown. To identify quantitative trait loci (QTLs) controlling radish morphological traits, we cultivated 94 F₄ and F₅ recombinant inbred lines derived from a cross between the rat-tail radish and the Japanese radish cultivar ‘Harufuku’ inbred lines. Eight morphological traits (ovule and seed numbers per silique, plant shape, pubescence and root formation) were measured for investigation. We constructed a map composed of 322 markers with a total length of 673.6 cM. The linkage groups were assigned to the radish chromosomes using disomic rape-radish chromosome-addition lines. On the map, eight and 10 QTLs were identified in 2008 and 2009, respectively. The chromosome-linkage group correspondence, the sequence-specific markers and the QTLs detected here will provide useful information for further genetic studies and for selection during radish breeding programs.     

Earthworms promote the reduction of Fusarium biomass and deoxynivalenol content in wheat straw under field conditions

A field experiment was conducted to elucidate ecosystem services provided by earthworms on the repression of phytopathogenic and toxinogenic fungi. The study focussed on decomposing Fusarium culmorum-infected and deoxynivalenol (DON)-contaminated wheat straw remaining on the soil surface as part in conservation tillage. Mesocosms were established in the topsoil of a winter wheat field located in Northern Germany, where conservation tillage has been practised for 20 years. Besides a non-earthworm treatment, two earthworm species were inoculated in the mesocosms either separately or combined: Lumbricus terrestris (anecic, detritivorous) and Aporrectodea caliginosa (endogeic, geophagous). The earthworms were exposed either to artificially Fusarium-infected wheat straw highly contaminated with DON or to non-infected straw serving as a control. The experiment was conducted during an eight week period after harvest from mid August to mid October. For both species, the artificially Fusarium-infected and DON-contaminated wheat straw was a more attractive food source than the non-infected control. In contrast to A. caliginosa, L. terrestris incorporated infected straw faster into the soil compared to control straw. Furthermore, the reduction of Fusarium biomass and DON concentration in wheat straw was significantly higher in the presence of L. terrestris than in treatments with A. caliginosa and without earthworms. Here, no significant differences could be measured between the Fusarium biomass and DON concentration in wheat straw. A. caliginosa seems not to be relevant for the reduction of Fusarium biomass and DON concentration. We concluded that amongst earthworms, anecic detritivorous species are the drivers to compensate possible negative consequences (like crop infection) of conservation tillage. They take an important role in the control of phytopathogenic and toxinogenic fungi surviving on plant residues and in the degradation of their mycotoxins.     

Characterization of Transgenic Pigs That Express Human Decay Accelerating Factor and Cell Membrane‐tethered Human Tissue Factor Pathway Inhibitor

The expression of human complement regulatory proteins (hCRP; hDAF, hCD59, and hMCP) in pig tissues has been suggested as one of strategies to overcome the hyperacute rejection (HAR) in pig‐to‐human transplantation. Expression of human tissue factor pathway inhibitor (hTFPI) in porcine endothelial cells has been suggested as a remedy to overcome microvascular thrombosis. To investigate the effects of these combined transgenes, we established transformed pig cells expressing human decay accelerating factor (hDAF) under the control of enhancer promoter (5′LTR‐PCMVIE), and the fusion protein (hTFPI/hCD4) consisting of the functional domains (K1 and K2) of hTFPI and membrane‐tethering domains (D3 and D4) of hCD4 under the control of PCMVIE. Transgenic pigs were generated with the transformed porcine cells through somatic cell nuclear transfer (SCNT) technology. Analysis of quantitative PCR and real‐time quantitative RT‐PCR showed that four copies of hDAF were integrated and 391 copies of hDAF mRNA expressed in the cells of the transgenic pig. The enhancing activity of 5′LTR was approximately 2 fold compared to CMVIE promoter only. The cell viability test showed that more than 80% of ear cells were viable in the presence of 50% human serum. The chromogenic substrate assay and immunocytochemical staining with tail cells showed that the TFPI activity of fusion protein was observed on the cell membrane. The membrane localization of hDAF and hTFPI proteins was observed by immunocytochemical staining, and the expression of transgenes in heart and liver tissues was also confirmed by immunohistochemistry.     

Successful backcrosses of somatic hybrids between Sinapis alba and Brassica oleracea with the Brassica oleracea parent

Somatic hybrids between Sinapis alba (2n= 24) and Brassica oleracea (2n= 18) have been backcrossed with the B. oleracea parent. Whereas backcrosses with the diploid B. oleracea parent were unsuccessful, 344 BC₁ seeds could be obtained from inter-valence crosses with tetraploid B. oleracea (2n= 4x= 36). The investigated 96 BC₁ plants segregated for morphological traits and for fertility. They were backcrossed with diploid B. oleracea or self-pollinated, depending on their male fertility. The BC₁F₂ and BC₂ progenies segregated well for the morphological traits. Disturbances were observed especially in the generative phase (flower development and pollen fertility). Both male fertile and male sterile BC₁F₂ and BC₂ plants were obtained and backcrossed or self-pollinated with the B. oleracea parent. The presence of either one of the parental or the cybrid organelle genomes was detected. In the progenies, a stable maternal inheritance of the organelle genome patterns was observed. Isozyme analyses revealed polymorphism for the leucine aminopeptidase (LAP) which was used for the identification of S. alba genes in the progenies. Cytological investigations showed a clear differentiation between the BC₁F₂ and BC₂ plants. Whereas the BC₁F₂ plants possess large numbers of chromosomes ranging from 34 to 40, the BC₂ material was strongly reduced to chromosome numbers ranging from 20 to 22. Preliminary investigation of the meiosis suggests the possibility of introgressions of S. alba-DNA into the B. oleracea genome.     

Regeneration of compacted soil aggregates by earthworm activity

Soil compaction is a problem of modern agriculture, caused by heavy machinery when used in unsuitable, especially moist, conditions. Some regeneration processes in compacted loess soil were studied in a field experiment near Relliehausen, at the edge of the Solling mountains in Lower Saxony, Germany. Conventional tillage (CT) and conservation tillage (CS) systems were compared. The compaction was induced by the use of different wheel loads. The influence of earthworms was determined by comparing soil aggregates and casts with respect to dry and moist porosity, swelling, and water stable aggregation. For visualisation of the microstructure, a scanning electron microscope was used. The casts were obtained from two earthworm species living for 6 months in the laboratory in monoliths, taken on the plots after the wheeling procedure. The casts showed 10-20% higher values for porosity and about 50% higher swelling values than comparable soil aggregates, while the relative water stability was ca. 10% lower. We conclude that casts are looser and less stable than aggregates from the soil the earthworms ingested. To show the ecological relevance of the changes in the casts, the cast production per hectare per year was calculated. It was especially high in the most loaded soil under CS with endogeic species.     

Earthworm effects on selected physical and chemical properties of soil aggregates

Summary Some physical and chemical properties of 1-to 2-mm aggregates obtained from casts and the burrow-wall material of the earthworm species Lumbricus terrestris, Aporrectodea longa, and Aporrectodea caliginosa were determined in order to show the effects of earthworms on the stabilization of soil aggregates. The results were compared with those of the natural soil from the Ap horizon of a Parabraunerde (Luvisol, FAO). Both the tensile strength and the water stability of aggregates from casts and burrow-wall material were reduced compared with those of the natural aggregates but were increased compared with those of remoulded aggregates. These results indicate that to a great extent existing bonds are destroyed by earthworm ingestion. Nevertheless, earthworm activities are advantageous for the stabilization of reformed aggregates. The coarse sand fraction is reduced by selective ingestion by earthworms. The organic C content is increased by 4.1–21.0% for burrow-wall material and by 21.2–43.0% for casts. The carbonate content of aggregates from casts and burrow-wall material of L. terrestris was reduced by more than 50%, while that of A. longa showed no noticeable changes and that of A. caliginosa was increased by more than 60%. The total content of polysaccharides was increased by 35–87% for casts and by 33–46% for the burrow-wall material of all earthworm species. The most frequently detected monosaccharides were glucose, galactose, and glucosamine. L. terrestris appeared to have the strongest effect on the interparticle bonding of the reformed aggregates, measured both as tensile strength and water stability, followed by A. longa and A. caliginosa.