Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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RNA-seq of Arabidopsis thaliana seedlings treated with Flg22 or Pep1

ABSTRACT: 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.

INSTRUMENT(S): Illumina HiSeq 2000

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Claude Becker

PROVIDER: E-MTAB-9838 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Experiment 1 Gene profiling upon biotic stress treatment Biological question : What are the genes (incuding microRNA precursors) that are differentially regulated upon flg22 treatment? Experiment description : 14-day old seedlings (Col-0 and fls2 KO) treated with 10uM of flg22 (flagellin- derived peptide) over a one hour timecourse. Experiment 2 Gene profiling in microRNA mutants Biological question : What are the genes (including miRNA precursors) that are differentially regulated in a set of microRNA mutants? Experiment description : Ler, dcl1-9, hen1-1 and hasty were grown for 12-day on MS solid medium ,seedlings were then transferred in MS liquid medium and harvested 2 days after. Experiment 3 Gene profiling in siRNA mutants Biological question : What are the genes (including miRNA precursors) that are differentially regulated in a set of siRNA mutants? Experiment description : Col-0, dcl2-1, dcl3-1 and rdr2-1 were grown for 12-day on MS solid medium ,seedlings were then transferred in MS liquid medium and harvested 2 days after. Experiment 4 Gene profiling in Agrobacterium-induced tumors and auxin-induced calli. Biological question : What are the genes (including miRNA precursors) that are differentially regulated in an Agrobacterium-induced tumors compared to auxin-induced calli? Experiment description : Col-0 plants were infected with wt Agrobacterium tumefaciens (strain A208) by stabbing the emerging stems. 18 days later, growing tumors were harvested as were the non-infected part of the stabbed-stem.In parallel, Col-0 seeds were germinated on Callus Induction Media (CIM) and 18 days later, calli were harvested. Experiment 5 Gene profiling in tasiRNA mutants Biological question : What are the genes (including miRNA precursors) that are differentially regulated in a set of tasiRNA mutants? Experiment description : Col-0, dcl4-1, rdr6-1 were grown for 12-day on MS solid medium ,seedlings were then transferred in MS liquid medium and harvested 2 days after. Experiment 6 Gene profiling upon biotic stress treatment Biological question : What are the genes (incuding microRNA precursors) that are differentially regulated upon flg22 treatment? Experiment description : 14-day old seedlings (Col-0 and fls2 KO) treated with 10uM of flg22 (flagellin- derived peptide) over a one hour timecourse.

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RNA-seq of Arabidopsis thaliana seedlings treated with Flg22 or Pep1

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