Project description:To determine transcriptome dynamics regulated by CPL3 during elicitation of immune responses, we performed RNA sequencing (RNA-seq) analysis of Col-0 WT and cpl3-3 mutant treated without or with flg22 for 30 min.
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:Purpose: The goals of this study are to analyze the transcriptome of Arabidopsis thaliana treated after flg22 and to find changes of glucosinolate metabolism genes after treatment. Methods: Total mRNA of 10 day wild-type Arabidopsis seedlings that were treated with and without flg22( final concentrations 1μmol/L) in 1/2 MS medium for 4h, were extracted respectively. Each sample was harvested in three independent biological replicates with equal weight and mixed ,subsequently sequencing. The sequence reads that passed quality filters were mapped to the Arabidopsis_thaliana genome (TAIR10.18).Then the mapping genes were used for the abundance and functional analysis. Results: We mapped about 45 million and 52 million sequence reads of control sample and treatment sample to the Arabidopsis_thaliana genome (TAIR10.18) and identified total 23,413 genes with Botiw/TopHat workflow. Comparison of the two samples showed 1,200 differentially expressed genes (DEGs), including 290 down-regulated and 910 up-regulated genes. The DEGs were associated with energy metabolism, amino acid metabolism and biosynthesis of secondary metabolites. After flg22 treatment, genes involved in indolic glucosinolate biosynthesis pathway were up-regulated significantly,which is further demonstrated by Real Time RT-PCR, while aliphatic glucosinolate pathway almost had no change, indicating the important role of indolic glucosinolates in plant defense responses. Conclusion: Our study provides the overall genetic resource of Arabidopsis_thaliana after treated by flg22 to date. These data will pave the way for further studies about deeply understand pathogen induced defense and the contribution of indolic glucosinolates.
Project description:Arabidopsis MPK11 is activated and plays a role in the flg22 sensing. Mitogen-activated protein kinases (MAPKs) mediate cellular signal transduction during stress responses, as well as diverse growth and developmental steps in eukaryotes. Pathogen infection or treatment with conserved pathogen-associated molecular patterns (PAMPs) such as the bacterial flagellin-derived flg22 peptide are known to activate three Arabidopsis thaliana MAPKs, MPK3, MPK4 and MPK6. Several stresses, including flg22 treatment, are known to increase MPK11 expression but activation of MPK11 has not been shown. Here, we show that MPK11 activity can indeed be increased through flg22 elicitation. Expression profiling using a small-scale microarray for defense-related genes revealed that cinnamyl alcohol dehyrogenase 5 (CAD5) requires MPK11 for full flg22-induced expression. An mpk11 mutant showed increased flg22-mediated growth inhibition but no altered susceptibility to Pseudomonas syringae, Botrytis cinerea or Alternaria brassicicola. A 24 DNA microarray study using toral RNA from an Arabidopsis mutant (mpk11 SALK049352) as well as wild type treated with water or flg22.
Project description:The experiments were performed to monitor dynamics in WRKY transcription factor abundancies upon treatment with flg22, a peptide derived from the bacterial flagella. Mock-treated or WT seedlings treated for 2 h with flg22 were used to prepare crude nuclear lysates. Then pull downs were performed with an anti-all-WRKY antiserum to enrich for WRKY transcription factor proteins prior to MS.