Magnus Simonsson

Rolling-circle replication of a high-copy BPV-1 plasmid

We investigated the replicating form of a bovine papillomavirus type 1 (BPV-1) deletion mutant by direct electron-microscopic analysis of low molecular weight cellular DNA fractions. The detection of viral plasmid DNA replication intermediates was facilitated by the isolation of a spontaneously transformed mouse cell subclone containing an unusually high viral genome copy number (approx. 1000 per cell), and by employing a slight modification of the Hirt fractionation procedure to reduce the level of contaminating linear chromosomal DNA fragments. We observed exclusively rolling-circle-type viral DNA replication intermediates, at a frequency of detection of approximately one replication intermediate per 200 monomeric circular viral DNA molecules. The demonstration of rolling-circles with longer-than-genome-length tails indicated that this high-copy viral plasmid was not subject to a strict once-per-cell-cycle mode of DNA replication. Our observations provide further evidence in favour of an alternative replication mode of the BPV-1 genome, and may help to explain earlier conflicting findings concerning the mechanism of stable BPV-1 plasmid copy-number-control.

Leaching of nutrients and major ions from an arable field with an unfertilized fallow as infield buffer zone

Abstract Leaching of nutrients and major ions from a tile-drained arable field was evaluated over a 25-year period (1980–2005). The soil, classified as a Gleyic Cambisol, received moderate applications of fertilizers. During later years the soil was more frequently under ley, and since 2002, an area of ravine (29% of the total field) has been managed as a permanent fallow with annual cutting without removing the grass material. A decrease in the concentration of nitrate nitrogen (NO3−N) in the drainwater was estimated in 1980–2005, based on non-parametric tests on concentrations flow-normalized with a robust fitting curve procedure. The average concentration of soluble reactive phosphorus after pre-filtration (RPf) of 0.030 mg l−l corresponded to a calculated degree of phosphorus saturation (DPS) of 6% in acid extract of ammonium lactate from the topsoil. Between 1995 and 2005, concentrations of suspended solids (SS), RP and total organic carbon (TOC) decreased significantly in the drainage water from the entire field. Throughout the entire period, there was a negative net accumulation of the major ions to the soil. The order of decrease in relative terms was calcium (Ca2+)>magnesium (Mg2+)>sulphate (SO4 2−)>chloride (Cl−)>potassium (K+). In addition, the soil phosphorus (P) balance was negative. A significant reduction in ion concentrations in the drainwater, including sodium (Na+) and hydrogen carbonate HCO3 −, was estimated. The order of reduction in relative terms was: SO4 2−>Mg2+>Ca2+>HCO3 −>Cl−>NO3 −>Na+>HPO4. Altogether these trends were equal to approximately 0.1 mmolc l−1. yr−1 of positively and negatively charged ions. Based on measured concentrations, decreasing trends in SO4 2− and Ca2+ were also observed in the shallow groundwater (3.6 m below the soil surface), while K+, Na+ and HCO3 − tended to increase. Trends of cations in deeper groundwater (5.8 m below the soil surface) were in some cases the opposite of the trends in the drainwater.

Structure liming enhances aggregate stability and gives varying crop responses on clayey soils

ABSTRACT It has been suggested that liming can improve soil structure and thereby decrease losses of particles and associated nutrients. In this study, two types of structure lime, slaked lime (Ca(OH)2) and a mixed product of calcium carbonate (CaCO3) and slaked lime (Ca(OH)2), were applied at three different rates in field trials on clayey soils (23%–40% clay). A combination of primary tillage and structure liming was also studied, in a split-plot trial on a clayey soil (25% clay). Aggregate (2–5 mm) stability, measured as reduction in turbidity (which is strongly correlated with losses of particulate phosphorus), was significantly increased with the highest application rates of both structure lime products. Aggregate size distribution was also improved with structure lime, creating a finer tilth in the seedbed. Yield response to structure lime was not consistent, with both negative and positive responses over the four-year study period. Positive yield responses can possibly be attributed to the finer tilth preventing evaporation in two dry growing seasons. Negative yield responses were probably an effect of impaired phosphorus availability associated with limited precipitation in May-July in 2011 and 2013. Two years after liming, soil pH levels were significantly elevated in plots with the highest application rate of structure lime, whereas no significant increases were found three years after liming. However, a lingering effect of liming was still detectable, as manganese concentration in barley grain was significantly lower in plots with the highest application rates of both structure lime products in the fourth study year. These results indicate that structure liming can be used as a measure to mitigate phosphorus losses from clayey soils, thereby preventing eutrophication of nearby waters. However, the yield response was varying and unpredictable and thus further investigations are needed to determine the circumstances in which field liming can act efficiently not only to prevent phosphorus losses, but also to ensure consistent yield increases.