Project description:Deep sequencing of mRNA from seven different tissues of Brassica oleracea Analysis of ploy(A)+ RNA of multiple different tissues of Brassica oleracea containing Bud, Callus, Root, Stem, Leaf, Flower and Silique.
Project description:Primary objectives: The primary objective is to investigate circulating tumor DNA (ctDNA) via deep sequencing for mutation detection and by whole genome sequencing for copy number analyses before start (baseline) with regorafenib and at defined time points during administration of regorafenib for treatment efficacy in colorectal cancer patients in terms of overall survival (OS).
Primary endpoints: circulating tumor DNA (ctDNA) via deep sequencing for mutation detection and by whole genome sequencing for copy number analyses before start (baseline) with regorafenib and at defined time points during administration of regorafenib for treatment efficacy in colorectal cancer patients in terms of overall survival (OS).
Project description:Fusarium oxysporum f. sp. conlutinans (Foc) is a serious root-invading and xylem-colonizing fungus that causes yellowing in Brassica oleracea. To comprehensively understand the interaction between F. oxysporum and B. oleracea, composition of the xylem sap proteome of the non-infected and Foc-infected plants was investigated in both resistant and susceptible cultivars using liquid chromatography-tandem mass spectrometry (LC-MS/MS) after in-solution digestion of xylem sap proteins. Whole genome sequencing of Foc was carried out and generated a predicted Foc protein database. The predicted Foc protein database was then combined with the public B. oleracea and B. rapa protein databases downloaded from Uniprot and used for protein identification. About 200 plant proteins were identified in the xylem sap of susceptible and resistant plants. Comparison between the non-infected and Foc-infected samples revealed that Foc infection causes changes to the protein composition in B. oleracea xylem sap where repressed proteins accounted for a greater proportion than those of induced in both the susceptible and resistant reactions. The analysis on the proteins with concentration change >=2 fold indicated a large portion of up- and down-regulated proteins were those acting on carbohydrates. Proteins with leucine-rich repeats and legume lectin domains were mainly induced in both resistant and susceptible system, so was the case of thaumatins. Twenty-five Foc proteins were identified in the infected xylem sap and ten of them were cysteine-containing secreted small proteins that are good candidates for virulence and/or avirulence effectors. The findings of differential response of protein contents in the xylem sap between the non-infected and Foc-infected samples as well as the Foc candidate effectors secreted in xylem provide valuable insights into B. oleracea-Foc interactions.
Project description:We investigated the expression profiles and genomic organization of PP2Cs-encoding genes in Brassica oleracea. Analysis of cDNA macroarray transcription profiles for Brassica oleracea and Arabidopsis thaliana revealed significant differences in the expression of a gene encoding protein phosphatase 2C, ABI1, a member of the group A PP2C. To gain insight into the ABA signaling network conservation in a model plant and its crop relatives group A PP2C genes in B. oleracea have been identified and functionally characterized. Twenty homologous sequences were identified as putative members of the group A PP2Cs (BolC.PP2Cs). Phylogenetic analysis revealed that the B. oleracea homologues are closely related to the particular members of the A. thaliana PP2C family. The genetic analysis has corroborated the presence of 2 to 3 copies for almost all of the PP2Cs examined, which corresponded to the unique genes in the A. thaliana genome. Gene expression analyses showed that among 15 PP2Cs-encoding genes studied in B.oleracea, BolC.ABI2, BolC.HAB1, BolC.HAB2.a-c, and BolC.PP2CA.a were drought-induced. However, in contrary to AtPP2Cs, only BolC.ABI1.a-b, BolC.ABI2 and BolC.PP2CA.a were ABA-responsive at the time points tested. Our results indicate that in B. oleracea PP2C-based drought stress signaling has evolved distinctly in comparison to A. thaliana. It is hypothesized that different reactions of particular B. oleracea PP2C genes to the water stress and ABA treatment may indicate lower conservation of their specificity in stress-induced reversible phosphorylation-based protein network operating in B. oleracea and A. thaliana.
Project description:We investigated the expression profiles and genomic organization of PP2Cs-encoding genes in Brassica oleracea. Analysis of cDNA macroarray transcription profiles for Brassica oleracea and Arabidopsis thaliana revealed significant differences in the expression of a gene encoding protein phosphatase 2C, ABI1, a member of the group A PP2C. To gain insight into the ABA signaling network conservation in a model plant and its crop relatives group A PP2C genes in B. oleracea have been identified and functionally characterized. Twenty homologous sequences were identified as putative members of the group A PP2Cs (BolC.PP2Cs). Phylogenetic analysis revealed that the B. oleracea homologues are closely related to the particular members of the A. thaliana PP2C family. The genetic analysis has corroborated the presence of 2 to 3 copies for almost all of the PP2Cs examined, which corresponded to the unique genes in the A. thaliana genome. Gene expression analyses showed that among 15 PP2Cs-encoding genes studied in B.oleracea, BolC.ABI2, BolC.HAB1, BolC.HAB2.a-c, and BolC.PP2CA.a were drought-induced. However, in contrary to AtPP2Cs, only BolC.ABI1.a-b, BolC.ABI2 and BolC.PP2CA.a were ABA-responsive at the time points tested. Our results indicate that in B. oleracea PP2C-based drought stress signaling has evolved distinctly in comparison to A. thaliana. It is hypothesized that different reactions of particular B. oleracea PP2C genes to the water stress and ABA treatment may indicate lower conservation of their specificity in stress-induced reversible phosphorylation-based protein network operating in B. oleracea and A. thaliana. For each genotype 7 samples were analysed; 4 controls and 3 samples extracted from drought-treated plants. The reliability and reproducibility of the macroarray analyses were ensured by using biological replicates in the experiment.