Project description:Trichomes of Arabidopsis thaliana have been broadly used to study cell development, cell differentiation and cell wall biogenesis. In this context, the exposed position, extraordinary size and characteristic morphology of trichomes featured particularly the exploration of trichome mutant phenotypes. However, trichome-specific biochemical or -omics analyses require a proper separation of trichomes from residual plant tissue. Thus, different strategies were proposed in the past for trichome isolation, which mostly rely on harsh conditions. To improve trichome-leaf separation, we revised a previously proposed method for isolating A. thaliana trichomes by optimizing the mechanical and biochemical specifications for trichome release. Furthermore, we introduced a density gradient centrifugation step to remove residual plant debris. We found that prolonged, yet mild seedling agitation increases the overall trichome yield by about 62% compared to the original protocol. We noticed that subsequent density gradient centrifugation further visually enhances trichome purity, which could be advantageous for downstream analyses. Histochemical and biochemical investigation of trichome cell wall composition indicated that gentle agitation during trichome release largely preserves trichome integrity. We used enriched and purified trichomes for proteomic analysis and present a reference data set of trichome-resident and -enriched proteins.
Project description:To understand how GTL1 regulates cell growth, we first identified its potential direct targets by the chromatin immunoprecipitation followed by the hybridization on an Affymetrix Arabidopsis Tiling 1.0R array (ChIP-chip). To enrich the genomic region bound by GTL1 in vivo, we harvested whole aerial parts of 12-day-old gtl1-1 plants complemented with the pGTL:GTL1:GFP constructs and immunoprecipitated the chromatin fragments associated with GTL1-GFP proteins using antibodies against GFP. After applying a cut-off P-values of 0.001of MAT (Model-based analysis of tiling array), we identified a total number of 3,900 putative immediate target genes that showed consistent binding by GTL1.
Project description:How bacteria from the microbiota modulate the physiology of its host is an important question to address. Previous work revealed that the metabolic status of Arabidopsis thaliana was crucial for the specific recruitment of Streptomycetaceae into the microbiota. Here, the Arabidopsis-Actinacidiphila interaction was further depicted by inoculating axenic Arabidopsis with Actinacidiphila cocklensis DSM 42063 or Actinacidiphila bryophytorum DSM 42138(previously named Streptomyces cocklensis and Streptomyces bryophytorum). We demonstrated that these two bacteria colonize A. thaliana wild-type plants, but their colonization efficiency was reduced in a chs5 mutant with defect in isoprenoid, phenylpropanoids and lipids synthesis. We observed that those bacteria affect the growth of the chs5 mutant but not of the wild-type plants. Using a mass spectrometry-based proteomic approach, we showed a modulation of the Arabidopsis proteome and in particular its components involved in photosynthesis or phytohormone homeostasis or perception by A. cocklensis and A. bryophytorum. This study unveils specific aspects of the Actinacidiphila-Arabidopsis interaction, which implies molecular processes impaired in the chs5 mutant and otherwise at play in the wild-type. More generally, this study highlights complex and distinct molecular interactions between Arabidopsis thaliana and bacteria belonging to the Actinacidiphila genus.
Project description:We use metabolite profiles of the model plant Arabidopsis thaliana measured on an UPLC-ESI/QqTOF-MS to evaluate uni- and multivariate statistical analysis of redundant features in compound spectra. Comparison was performed between the wild-type Col-0 and the 90.32 mutant. The mutant is a transposon based activation tagged A. th. line from the TAMARA population Schneider et al. [2005]. This particular mutant has an over-expression of the AT5G55880 - AT5G55890 genetic region with unknown function.
Project description:The oxt6 mutant is an oxidative stress-tolerant Arabidopsis mutant that is deficient in a polyadenylation factor subunit. Expression analysis suggests that impaired poly(A) site choice is responsible for the stress-tolerant phenotype. We used microarrays to understand the link between the polyadenylation defect and stress tolerance. Keywords: mutant, wild-type, complemented line comparison