Project description:We have combined a modified protein extraction method, heat/thaw/phenol/chloroform (HTPC), with the established Surfactant extraction method to identify proteins from Park Grass Experiment (PGE) soil, which has an extensively sequenced microbial database.
Project description:Background: The soil environment is responsible for sustaining most terrestrial plant life on earth, yet we know surprisingly little about the important functions carried out by diverse microbial communities in soil. Soil microbes that inhabit the channels of decaying root systems, the detritusphere, are likely to be essential for plant growth and health, as these channels are the preferred locations of new root growth. Understanding the microbial metagenome of the detritusphere and how it responds to agricultural management such as crop rotations and soil tillage will be vital for improving global food production. Methods: The rhizosphere soils of wheat and chickpea growing under + and - decaying root were collected for metagenomics sequencing. A gene catalogue was established by de novo assembling metagenomic sequencing. Genes abundance was compared between bulk soil and rhizosphere soils under different treatments. Conclusions: The study describes the diversity and functional capacity of a high-quality soil microbial metagenome. The results demonstrate the contribution of the microbiome from decaying root in determining the metagenome of developing root systems, which is fundamental to plant growth, since roots preferentially inhabit previous root channels. Modifications in root microbial function through soil management, can ultimately govern plant health, productivity and food security.
Project description:We used an immature mouse T cell line engineered to express a biotinylated form of the cleaved form of Notch1 (ICN1). ICN1-bound sites were precipitated with streptavidin-coated beads and subjected to ChIP-sequencing. Beko cells correspond to a spontaneous T lymphoma immature cell line derived from a TCRb deficient mouse. These cells were engineered to express a biotin-tagged-ICN1 and the bacterial biotin ligase BirA (Bio-ICN1) or just BirA as a control (Bio). Chromatin from both cell lines was subjected to strepatavidin-mediated precipitation and subjected to sequencing with the Illumina GAII sequencer as single end 36 base pair reads.
Project description:Grass carp (Ctenopharyngodon idellus), the world’s largest aquaculture fish species, exhibits superior growth in females compared to males. However, the lengthy sexual maturation period of four to five years poses a significant obstacle to the genetic reproduction and breeding of grass carp. Consequently, classical methods such as gonadogenesis or sex reversal through steroid treatment, employed for breeding all-female grass carp, demand considerable time and effort. In this study, we developed an super-fast breeding strategy for generating all-female grass carp in a total of half a year, using a surrogate production method. We first characterized grass carp female germline stem cells (GSCs) from genetic female juveniles at three months post-fertilization (mpf). The female GSCs with XX chromosomes were then transplanted into germ cell-depleted zebrafish larvae at five days post-fertilization (dpf). The transplanted grass carp XX germ cells underwent rapid spermatogenesis in the zebrafish recipient. At three months after transplantation, all zebrafish recipients had developed into males capable of producing the all-X sperm of the grass carp. By using these sperm to fertilize wildtype grass carp eggs, we successfully produced an all-female grass carp offspring. This groundbreaking achievement highlights the potential of surrogate production in the genetic breeding of valuable fish species, and opens a new avenue for advancing genetic breeding in aquaculture.
Project description:We measured transcriptional profiles of individuals of Andropogon gerardii, a C4 grass native to North American grasslands, in a field experiment in which both temperature and precipitation have been manipulated to simulate key aspects of forecasted climate change.