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Deforestation of tropical rainforests is a major land use change that alters terrestrial biogeochemical cycling at local to global scales. Deforestation and subsequent reforestation are likely to impact soil phosphorus (P) cycling, which in P-limited ecosystems such as the Amazon basin has implications for long-term productivity. We used a 100-year replicated observational chronosequence of primary forest conversion to pasture, as well as a 13-year-old secondary forest, to test land use change and duration effects on soil P dynamics in the Amazon basin. By combining sequential extraction and P K-edge X-ray absorption near edge structure (XANES) spectroscopy with soil phosphatase activity assays, we assessed pools and process rates of P cycling in surface soils (0-10 cm depth). Deforestation caused increases in total P (135-398 mg kg-1 ), total organic P (Po ) (19-168 mg kg-1 ), and total inorganic P (Pi ) (30-113 mg kg-1 ) fractions in surface soils with pasture age, with concomitant increases in Pi fractions corroborated by sequential fractionation and XANES spectroscopy. Soil non-labile Po (10-148 mg kg-1 ) increased disproportionately compared to labile Po (from 4-5 to 7-13 mg kg-1 ). Soil phosphomonoesterase and phosphodiesterase binding affinity (Km ) decreased while the specificity constant (Ka ) increased by 83%-159% in 39-100y pastures. Soil P pools and process rates reverted to magnitudes similar to primary forests within 13 years of pasture abandonment. However, the relatively short but representative pre-abandonment pasture duration of our secondary forest may not have entailed significant deforestation effects on soil P cycling, highlighting the need to consider both pasture duration and reforestation age in evaluations of Amazon land use legacies. Although the space-for-time substitution design can entail variation in the initial soil P pools due to atmospheric P deposition, soil properties, and/or primary forest growth, the trend of P pools and process rates with pasture age still provides valuable insights.

Correct reprogramming of the DLK1-DIO3 imprinted region is critical for the development of cloned animals. However, in pigs, the imprinting and regulation of the DLK1-DIO3 region has not been systematically analyzed. The objective of this study was to investigate the imprinting status and methylation regulation of the DLK1-DIO3 region in wild-type and cloned neonatal pigs. We mapped the imprinting control region, IG-DMR, by homologous alignment and validated it in sperm, oocytes, fibroblasts, and parthenogenetic embryos. Subsequently, single nucleotide polymorphism-based sequencing and bisulfite sequencing polymerase chain reaction were conducted to analyze imprinting and methylation in different types of fibroblasts, as well as wild-type and cloned neonatal pigs. The results showed that Somatic cell nuclear transfer (SCNT) resulted in hypermethylation of the IG-DMR and aberrant gene expression in the DLK1-DIO3 region. Similar to wild-type pigs, imprinted expression and methylation were observed in the surviving cloned pigs, whereas in dead cloned pigs, the IG-DMR was hypermethylated and the expression of GTL2 was nearly undetectable. Our study reveals that abnormal imprinting of the DLK1-DIO3 region occurs in cloned pigs, which provides a theoretical basis for improving the cloning efficiency by gene editing to correct abnormal imprinting.

Copper (Cu) products, including copper oxide nanoparticles (nCuO), are critically important agricultural fungicides and algaecides. Foliar application onto crops and subsequent aerosol drift of these Cu products, especially nCuO, on to soil may alter nutrient cycling and microbial communities in both managed and unmanaged environments. We measured the influence of land use on soil microbial biomass and respiration in response to the addition of nCuO to an alluvial soil. Different land uses included grassland, forest and both organic and conventional managed row crops. Soil samples were amended with 1000 mg Cu per kg soil as CuCl2, 16 nm CuO (16nCuO), 42 nm CuO (42nCuO), and larger than nanoparticle sized bulk CuO (bCuO). Copper availability immediately increased in all soils following Cu addition in the order of CuCl2 > 16nCuO > 42nCuO > bCuO. After 70 days Cu availability was diminished across land uses and lowest in soils treated with bCuO. Using X-ray absorption near edge structure (XANES) spectroscopy, we determined that the relatively high availability of Cu after treatment with nanoparticle sized CuO was due to the dissolution of CuO particles and subsequent adsorption by soil materials. Respiration, an indicator of microbial activity, was suppressed by Cu additions, especially CuCl2. Copper effects on soil microbial biomass were sensitive to land use. In agricultural soils, microbial biomass was unaltered by Cu form, regardless of concentration, whereas in unmanaged soils, it decreased following exposure to CuCl2 and 42nCuO. Our results suggest that land use history has little impact on Cu chemical fate in soils, but strongly modulates microbial response to Cu exposure. These results are especially important for organic agricultural systems where copper fungicides are widely used but may suppress microbial mineralization of nutrients from soil organic matter.