|
| Status |
Public on Aug 01, 2009 |
| Title |
WRKY23-RNAi_Mmt-inoculated_rep3 |
| Sample type |
RNA |
| |
|
| Source name |
WRKY23-RNAi leaf discs, Mmt-inoculated
|
| Organism |
Populus tremula x Populus alba |
| Characteristics |
tissue: Leaf discs from fully expanded leaf LPI9 (leaf plastochron index)
genotype: WRKY23-RNAi
|
| Treatment protocol |
Leaf discs (2-cm diameter) prepared from leaf LPI9 (leaf plastochron index) of 1-m tall plants were inoculated on the abaxial side with Melampsora medusae f. sp. tremuloidae (Mmt) urediniospores suspended in 0.01% tween at a density of 500 spores per cm2. After inoculation, discs were kept on wet paper in petri dishes and incubated in a growth chamber at 18°C with a 16-h photoperiod until sample collection at 5 dpi.
|
| Growth protocol |
Wildtype and WRKY23-RNAi plants of Populus tremula x P. alba clone INRA 717 1-B4 were grown in a mixture of peatmoss, vermiculite and turface (4:2:1) with a photoperiod of 16-h day/8-h night at 24°C/20°C. Melampsora medusae f. sp. tremuloidae (Mmt) were maintained on Populus tremuloides according to Boyle et al., 2005 [Boyle B, Hamelin RC, Seguin A (2005) In vivo monitoring of obligate biotrophic pathogen growth by kinetic PCR. Applied and Environmental Microbiology 71: 1546-1552].
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA were extracted from frozen leaf tissues according to Chang et al., 1993 [Chang S, Puryear J, Cairney J (1993) A simple and efficient method for isolating RNA from pine trees. Plant Molecular Biology Reporter 11: 113-116] and were purified on RNeasy columns (RNeasy Plant Mini Kit, Qiagen). Following a DNase I treatment (Qiagen), 10 μg of total RNA from each biological sample were used for each microarray experiment.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 10 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Poplar Genome Array. GeneChips were washed and stained in the Fluidics protocol EukGE_WS2v5_450.
|
| Scan protocol |
GeneChip scanner.
|
| Description |
Gene expression data from infected tissues with Melampsora medusae f. sp. tremuloidae (Mmt).
|
| Data processing |
Robust Multichip Average (RMA) via FlexArray software package (http://genomequebec.mcgill.ca/FlexArray).
|
| |
|
| Submission date |
Jun 03, 2009 |
| Last update date |
Jul 28, 2009 |
| Contact name |
Armand Séguin |
| Organization name |
Natural Resources Canada
|
| Department |
Laurentian Forestry Centre
|
| Lab |
Armand Séguin
|
| Street address |
1055 du P.E.P.S., PO Box 3800
|
| City |
Sainte-Foy |
| State/province |
Quebec |
| ZIP/Postal code |
G1V 4C7 |
| Country |
Canada |
| |
|
| Platform ID |
GPL4359 |
| Series (2) |
| GSE16419 |
Expression profiling and functional analysis of poplar WRKY23 reveals a regulatory role in defense: WRKY23-RNAi |
| GSE16420 |
Expression profiling and functional analysis of poplar WRKY23 reveals a regulatory role in defense |
|
