Four years of litter input manipulation changes soil microbial characteristics in a temperate mixed forest

Aboveground litter not only is an important source of nutrients to soil microbes but also regulates the microclimate in topsoil. How the changes in aboveground litter quantity would affect the microbial biogeochemical cycles is still unclear. Here we conducted a litter input manipulation experiment in a temperate mixed forest to investigate how different amounts of litter input affect soil organic carbon (SOC) and soil respiration via their regulation on soil microbes. We found that although neither SOC stock nor soil CO₂ efflux was affected by litter manipulation, soil microbial characteristics had responded after four years of litter addition or removal treatments. Microbial biomass carbon (MBC) in the O horizon was higher in litter addition plots than in litter removal plots as a result of the changed availability of labile C under litter treatments. Both double litter and no litter treatments changed microbial compositions, which was probably due to the increased soil pH in no litter treatment and the increased labile C in double litter treatment. The null change in soil respiration could be attributed to the offset between the negative effect of decreased substrate and the positive effect of increased temperature on soil respiration in litter removal plots. Due to the important role of soil microbes in carbon cycling, the altered microbial properties under litter manipulation treatments suggested the inevitable changes in biogeochemical cycling in the long run and call for long-term studies on SOC dynamics in the future.

     

Triticale and barley microspore embryogenesis induction requires both reactive oxygen species generation and efficient system of antioxidative defence

Plant Cell, Tissue and Organ Culture (PCTOC) - The effectiveness of microspore embryogenesis (ME) is determined by a complex network of internal and environmental factors. In the present study on...     

RPA2 winged-helix domain facilitates UNG-mediated removal of uracil from ssDNA; implications for repair of mutagenic uracil at the replication fork

Uracil occurs at replication forks via misincorporation of deoxyuridine monophosphate (dUMP) or via deamination of existing cytosines, which occurs 2–3 orders of magnitude faster in ssDNA than in dsDNA and is 100% miscoding. Tethering of UNG2 to proliferating cell nuclear antigen (PCNA) allows rapid post-replicative removal of misincorporated uracil, but potential ‘pre-replicative’ removal of deaminated cytosines in ssDNA has been questioned since this could mediate mutagenic translesion synthesis and induction of double-strand breaks. Here, we demonstrate that uracil-DNA glycosylase (UNG), but not SMUG1 efficiently excises uracil from replication protein A (RPA)-coated ssDNA and that this depends on functional interaction between the flexible winged-helix (WH) domain of RPA2 and the N-terminal RPA-binding helix in UNG. This functional interaction is promoted by mono-ubiquitination and diminished by cell-cycle regulated phosphorylations on UNG. Six other human proteins bind the RPA2-WH domain, all of which are involved in DNA repair and replication fork remodelling. Based on this and the recent discovery of the AP site crosslinking protein HMCES, we propose an integrated model in which templated repair of uracil and potentially other mutagenic base lesions in ssDNA at the replication fork, is orchestrated by RPA. The UNG:RPA2-WH interaction may also play a role in adaptive immunity by promoting efficient excision of AID-induced uracils in transcribed immunoglobulin loci.     

H4K20me1 and H3K27me3 are concurrently loaded onto the inactive X chromosome but dispensable for inducing gene silencing

During X chromosome inactivation (XCI), in female placental mammals, gene silencing is initiated by the Xist long non‐coding RNA. Xist accumulation at the X leads to enrichment of specific chromatin marks, including PRC2‐dependent H3K27me3 and SETD8‐dependent H4K20me1. However, the dynamics of this process in relation to Xist RNA accumulation remains unknown as is the involvement of H4K20me1 in initiating gene silencing. To follow XCI dynamics in living cells, we developed a genetically encoded, H3K27me3‐specific intracellular antibody or H3K27me3‐mintbody. By combining live‐cell imaging of H3K27me3, H4K20me1, the X chromosome and Xist RNA, with ChIP‐seq analysis we uncover concurrent accumulation of both marks during XCI, albeit with distinct genomic distributions. Furthermore, using a Xist B and C repeat mutant, which still shows gene silencing on the X but not H3K27me3 deposition, we also find a complete lack of H4K20me1 enrichment. This demonstrates that H4K20me1 is dispensable for the initiation of gene silencing, although it may have a role in the chromatin compaction that characterises facultative heterochromatin.