Project description:This study evaluates the transcriptome of Arabidopsis thaliana roots exposed to the MAMP flg22 in the presence of a 35-member bacterial SynCom
Project description:To study the effect of microbe-associated molecular pattern (MAMP) treatment (flg22) on the phosphorylation of nuclear proteins, we treated Arabidopsis plants with flg22 and after isolation of nuclear proteins, we enriched for phosphopeptides and then carried out LC-MS/MS analyses. We used statistical analysis tools to identify differentially phosphorylated proteins in response to MAMP treatment in the mock and treated samples.
Project description:Genome-wide gene expression analysis of the effects of PARP inhibitor (3AB) and, separately, a parg1 knockout, on early microbe-associate molecular pattern (MAMP)-induced gene expression in the plant basal defense response. Arabidopsis thaliana wild-type (Col-0), 3AB-treated and parg1-2 T-DNA knockout plants responding to the MAMP elicitors flg22 or elf18 were studied. The mutant and PARP inhibitors analyzed in this study are further described in Adams-Phillips, L.,C., Briggs, A.,G., Bent, A., F. 2010. Disruption of poly(ADP-ribosyl)ation mechanisms alters responses of Arabidopsis to biotic stress. Planty Physiol. 152(1):267-80 DOI: 10.1104/pp.109.148049.
Project description:This study evaluates the transcriptome of Arabidopsis thaliana seedlings chronically exposed to the hormone Methyl Jasmonate (MeJA) or to the bacterial elicitor flg22 (a 22-amino acid peptide from flagellin). Treatments were performed under high and low phosphate availability using wild-type Col-0 plants and the phr1 phl1 mutant.
Project description:In this research a high-throughput RNA sequencing based transcriptome analysis technique (RNA-Seq) was used to evaluate differentially expressed genes (DEGs) in the wild type Arabidopsis seedling in response to flg22, a well-known microbe-associated molecular patterns (MAMP), and AtPep1, a well-known peptide representing an endogenous damage-associated molecular patterns (DAMP). The results of our study revealed that 1895 (1634 up-regulated and 261 down-regulated) and 2271 (1706 up-regulated and 565 down-regulated) significant differentially expressed genes in response to flg22 and AtPep1 treatment, respectively. Among significant DEGs, we observed that a number of hitherto overlooked genes have been found to be induced upon treatment with either flg22 or with AtPep1, indicating their possible involvement in innate immunity. Here, we characterized two of them, namely PP2-B13 and ACLP1. PP2-B13 contains an F-box domain and shows similarity to carbohydrate binding proteins. ACLP1 is a protein of unknown function with highest similarity to actin cross linking proteins and includes a fascin domain. Using qPCR, we verified that the genes encoding PP2-B13, and ACLP1 were highly induced upon treatment of leaf disks with flg22. We obtained T-DNA insertion mutants and generated homozygous mutant lines. None of the mutants showed a phenotype in the absence of infection. pp2-b13 and aclp1 mutants showed an increased susceptibility to infection by the virulent pathogen Pseudomomas syringae pv tomato mutant hrcC-, as evidenced by an increased growth of the pathogen in planta. Further we present evidence that aclp1 was deficient in ethylene production upon flg22 treatment, while pp2-b13, was deficient in ROS production. In conclusion, the products of these genes contribute to plant immunity against bacterial pathogens, although there is currently no clue for their mechanism of action. The results from this research provide new information to a better understanding of the immune system in Arabidopsis.
Project description:Salicylic acid (SA)-induced defense responses are important factors during effector triggered immunity and microbe-associated molecular pattern (MAMP)-induced immunity in plants. This article presents evidence that a member of the Arabidopsis CBP60 gene family, CBP60g, contributes to MAMP-triggered SA accumulation. CBP60g is inducible by both pathogen and MAMP treatments. Pseudomonas syringae growth is enhanced in cbp60g mutants. Expression profiles of a cbp60g mutant after MAMP treatment are similar to those of sid2 and pad4, suggesting a defect in SA signaling. Accordingly, cbp60g mutants accumulate less SA when treated with the MAMP flg22 or a P. syringae hrcC strain that activates MAMP signaling. MAMP-induced production of reactive oxygen species and callose deposition are unaffected in cbp60g mutants. CBP60g is a calmodulin-binding protein with a calmodulin-binding domain located near the N-terminus. Calmodulin binding is dependent on Ca2+. Mutations in CBP60g that abolish calmodulin binding prevent complementation of the SA production and bacterial growth defects of cbp60g mutants, indicating that calmodulin binding is essential for the function of CBP60g in defense signaling. These studies show that CBP60g constitutes a calmodulin-dependent link between MAMP recognition and SA accumulation that is important for resistance to P. syringae. This experiment consists of three biological replicates. For each genotype, two leaves per plant were pooled from three pots to prepare total RNA.