root - Modified Fahraeus medium (in vitro), 25 degres, 16h, 80-100 µmol.m-2.s-1
Extracted molecule
total RNA
Extraction protocol
DMSO_H2O2:12ug.
Label
Biotin
Label protocol
labelling Biotin direct, amplification=yes, cRNA 20 ug.
Hybridization protocol
labelled extract quantity: 15ug
Scan protocol
GCOS, Biotin:pmt voltage 570nm,650V,laser power 1
Description
The involvement of ROS in the legume – Rhizobium symbiotic interaction has been highlighted (Santos et al., 2001; Rubio et al., 2004). This interaction is characterized by the formation of a new organ on the root, the nodule and by the penetration, in parallel, of the bacteria into the root tissue via an infection thread (IT) (Parniske and Downie, 2003; Gage, 2004). H2O2 production has been shown in ITs during the Medicago – Sinorhizobium meliloti interaction (Santos et al., 2001). Moreover, S. meliloti mutants impaired in H2O2 detoxification mechanism possess in planta symbiotic phenotypes. As H2O2 production is known to orchestrate plant gene expression, our goal is focused on identifying the H2O2 regulated genes in the symbiotic process. We focus our analysis in the M. truncatula transcriptome as we are characterising the S. meliloti H2O2 transcriptome.
Data processing
The data were normalized with the gcrma algorithm (Irizarry et al., 2003), available in the Bioconductor package (Gentleman and Carey, 2002).
