Functional definition of the mutation cluster region of adenomatous polyposis coli in colorectal tumours

The mutation cluster region (MCR) of adenomatous polyposis coli (APC) is located within the central part of the open reading frame, overlapping with the region encoding the 20 amino acid repeats (20R) that are β-catenin-binding sites. Each mutation in the MCR leads to the synthesis of a truncated APC product expressed in a colorectal tumour. The MCR extends from the 3′ border of the first 20R coding region to approximately the middle of the third 20R coding region, reflecting both positive and negative selections of the N- and C-terminal halves of the APC protein in colon cancer cells, respectively. In contrast, the second 20R escapes selection and can be either included or excluded from the truncated APC products found in colon cancer cells. To specify the functional outcome of the selection of the mutations, we investigated the β-catenin binding capacity of the first three 20R in N-terminal APC fragments. We found in co-immunoprecipitation and intracellular co-localization experiments that the second 20R is lacking any β-catenin binding activity. Similarly, we also show that the tumour-associated truncations abolish the interaction of β-catenin with the third 20R. Thus, our data provide a functional definition of the MCR: the APC fragments typical of colon cancer are selected for the presence of a single functional 20R, the first one, and are therefore equivalent relative to β-catenin bindin

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Isolation and Molecular Characterization of Chitinase-Deficient Bacillus licheniformis Strains Capable of Deproteinization of Shrimp Shell Waste To Obtain Highly Viscous Chitin

Proteolytic but chitinase-deficient microbial cultures were isolated from shrimp shell waste and characterized. The most efficient isolate was found to be a mixed culture consisting of two Bacillus licheniformis strains, which were first determined microscopically and physiologically. Molecular characterization was carried out by sequencing the 16S rRNA gene of both strains. According to the residual protein and ash content, the chitin obtained by fermentation of such a mixed culture was found to be comparable to a commercially available, chemically processed product. However, the strikingly high viscosity (80 versus 10 mPa of the commercially available sample) indicates its superior quality. The two strains differed in colony morphology and in their secretion capabilities for degradative extracellular enzymes. Sequencing of the loci encoding amylase, cellulase, chitinases, and proteases, as well as the degS/degU operon, which is instrumental in the regulation of degradative enzymes, and the pga operon, which is responsible for polyglutamic acid production, revealed no differences. However, a frameshift mutation in chiA, encoding a chitinase, was validated for both strains, providing an explanation for the ascertained absence of chitinolytic activities and the concomitant possibility of producing highly viscous chitin in a fermentational deproteinization process

Nitrogen fertilization strategies to reduce the risk of nitrate leaching in open field cultivation of spinach (Spinacia oleracea L.)

Background Spinach is a nitrogen (N) demanding crop with a weekly N uptake of up to 60 kg ha–1. Consequently, a high N supply is required, which can temporarily lead to high quantities of nitrate (NO3–) being at risk of leaching. Aims The objective of this study was to develop a N fertilization approach to reduce the risk of NO3– leaching in field-grown spinach production without adversely affecting crop yield and quality at an early and late harvest stage. Methods Ten fertilization trials were conducted to compare different base fertilization rates and splits of top dressings. For top dressings, granulated fertilizers or foliar sprays were used. In a further treatment, N supply was reduced by withholding the second top dressing of 50–70 kg ha−1. Results Nitrate concentration at risk of leaching was considerably reduced by decreasing the base fertilizer rate as well as by splitting the top dressing. However, at an early harvest stage, total aboveground dry mass was reduced by, on average, 6% by these measures across all seasons. In contrast, at a later harvest stage, spinach was less affected by the fertilizer schedule. Urea foliar sprays proved to be insufficient in promoting plant growth and caused leaf necrosis. A reduced N supply led to impaired plant growth and yellowish leaves in both spring and winter. Conclusions Base N fertilization of spinach is only required in spring, but not in other seasons. Despite slight yield reduction, the top dressing should be split to reduce the risk of NO3− leaching after an early harvest

Ernterückstandsmanagement nach Herbstspinat zur Minderung von Stickstoffverlusten über die Sickerwasserperiode im Winter

In open-field vegetable production, high quantities of soil mineral nitrogen (Nmin) and N-rich crop residues often remain in the field at harvest. After the harvest of crops in autumn, this N can lead to considerable nitrate (NO3-) losses during the subsequent winter leaching period. In four field trials, different tillage depths (3–4, 10, 30 cm) and dates (early autumn, late autumn, early spring) were investigated to reduce N losses after growing spinach in the autumn. In a further treatment, the nitrification inhibitor 3,4-Dimethylpyrazole phosphate (DMPP) was directly applied to the crop residues. Potential N losses were calculated by a balance sheet approach based on Nmin concentration (0–90 cm), measured N mineralization and N uptake by catch crops. By postponing the tillage date from early to late autumn or spring, resprouting spinach stubbles acted as a catch crop, reducing N losses by up to 61 kg ha-1. However, if the spinach biomass collapsed, the N losses increased by up to 33 kg ha-1 even without tillage. The application of DMPP as well as the tillage depth were less effective. Overall, postponing tillage to spring seems to be the most promising approach for reducing N losses during the off-season

Iodine biofortification of field-grown strawberries – Approaches and their limitations

The potential of iodine biofortification in strawberry fruits by means of soil and foliar fertilization was investigated in three field experiments and a preliminary phytotoxicity test in the greenhouse. In the main experiment iodine was applied by one-time potassium iodate soil drenches two weeks after planting or, alternatively, by using potassium iodide foliar sprays from the beginning of flowering. Beside the iodine accumulation in fruits, effects on crop yield and quality were determined. The soil fertilization resulted in a relatively low iodine accumulation in strawberry fruits, probably because the concentration of phytoavailable iodine in the soil rapidly decreased after its application. A markedly higher iodine content in fruits was achieved when it was aerially applied, either by a single treatment shortly before harvest or by repeated sprays during the flowering period. Yield, firmness and total acidity concentration of strawberry fruits were not significantly affected by any of the tested iodine applications. However, as a result of repeated foliar sprays the concentration of soluble solids in fruits was slightly diminished. Attemps to substantially increase the iodine content in fruits of strawberry plants cultivated in the second and third year failed, even following frequent sprays. In conclusion the results of this study suggest that only a relatively small proportion of exogenously applied iodine enters the fruits of field-grown strawberries due to its strong retention in soil and low phloem mobility in plants

Crop Residue Management Strategies to Reduce Nitrogen Losses during the Winter Leaching Period after Autumn Spinach Harvest

In open-field vegetable production, high quantities of soil mineral nitrogen (Nmin) and N-rich crop residues often remain in the field at harvest. After the harvest of crops in autumn, this N can lead to considerable nitrate (NO3−) losses during the subsequent winter leaching period. In four field trials, different tillage depths (3–4, 10, 30 cm) and dates (early autumn, late autumn, early spring) were investigated to reduce N losses after growing spinach in the autumn. In a further treatment, the nitrification inhibitor 3,4-Dimethylpyrazole phosphate (DMPP) was directly applied to the crop residues. Potential N losses were calculated by a balance sheet approach based on Nmin concentration (0–90 cm), measured N mineralization and N uptake by catch crops. By postponing the tillage date from early to late autumn or spring, resprouting spinach stubbles acted as a catch crop, reducing N losses by up to 61 kg ha−1. However, if the spinach biomass collapsed, the N losses increased by up to 33 kg ha−1 even without tillage. The application of DMPP as well as the tillage depth were less effective. Overall, postponing tillage to spring seems to be the most promising approach for reducing N losses during the off-season

Crop residue management strategies to reduce nitrogen losses during the winter leaching period after autumn spinach harvest

In open-field vegetable production, high quantities of soil mineral nitrogen (N-min) and N-rich crop residues often remain in the field at harvest. After the harvest of crops in autumn, this N can lead to considerable nitrate (NO3- ) losses during the subsequent winter leaching period. In four field trials, different tillage depths (3-4, 10, 30 cm) and dates (early autumn, late autumn, early spring) were investigated to reduce N losses after growing spinach in the autumn. In a further treatment, the nitrification inhibitor 3,4-Dimethylpyrazole phosphate (DMPP) was directly applied to the crop residues. Potential N losses were calculated by a balance sheet approach based on N-min concentration (0-90 cm), measured N mineralization and N uptake by catch crops. By postponing the tillage date from early to late autumn or spring, resprouting spinach stubbles acted as a catch crop, reducing N losses by up to 61 kg ha(-1). However, if the spinach biomass collapsed, the N losses increased by up to 33 kg ha(-1) even without tillage. The application of DMPP as well as the tillage depth were less effective. Overall, postponing tillage to spring seems to be the most promising approach for reducing N losses during the off-season