Transcription profiling of Populus euphratica leaves and roots in response to salinity

ABSTRACT: Rooted plantlets of Populus euphratica were transferred to aerated hydroculture with Long Ashton nutrient solution, grown for 3.5 months at 22°C, 150 µmol m-2 s-1 photosynthetically active radiation with a photoperiod of 16h light. After reaching an average height of 0.83 m, the media was supplemented with 150 mM NaCl (Salt-treated) or maintained under control conditions (control). During the experimental phase, light was continuously supplied to avoid interference of light/dark transitions.
Roots and leaves of six plants were harvested separately after 3h, 6h, 12h, and 24h of salt exposure and of controls at the corresponding time points (= 48 plants per experiment). The material of each harvest of 6 plants was pooled, yielding one leaf and one root sample of controls and one leaf and one root sample of salt-treated plants, respectively (=16 samples per experiment).
The experiment was performed three times yielding three biological replications (=48 samples in total).
After extraction of RNA and cDNA synthesis, each extract was labelled with Cy3 and Cy5, respectively (yielding 96 labelled extracts). Extracts from e.g. 3h-salt-treated leaf exp. 1 (cy3) was hybridized with 3h-control leaf exp.1 (cy5) with microarrays (UHPB-P.euphratica-10K-5; A-MEXP-93)containing unique 7,662 ESTs of P. euphratica from stress-induced cDNA libraries (Brosche et al., Genome Biology 2005, 6:R101). Dye swap was perfomed, thus usually yielding 6 arrays per timepoint and plant tissue (3 biological, 3 technical replicates). In some cases hydridizations were not successfull. Therefore, for roots harvested at timepoints 12h and 24 h only 4 arrays per timepoint were used. For roots harvested after 3h an additional mixed sample of all 3 biological replicates was performed. One leaf array at 6 h was not sucessfull. Therefore, residual labelled extract from salt treated plants of exp1 was hydridized with controls at 6h from exp.2.

ORGANISM(S): Populus euphratica

SUBMITTER: Andrea Polle 

PROVIDER: E-MEXP-2095 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress