Affymetrix GeneChip Wheat Genome Array [wheat]

Host-selective toxins of Pyrenophora tritici-repentis induce common responses associated with host susceptibility.

Host-Selective Toxins of Pyrenophora tritici-repentis Induce Common Responses Associated with Host Susceptibility

rawData

Water and ToxB-infiltrated leaves of sensitive cultivar Katepwa were collected at 0,3,9,14,24,48 hours post infiltration (hpi), total RNA was extracted and processed. Expression levels were obtained using Affymetrix GeneChip Wheat Genome Array

Time

Treatment

Transcription profiling by array of wheat leaves in response to the fungal toxin ToxB from Pyrenophora tritici-repentis

Transcriptional time course analysis of Wheat leaves in response to exposure to the fungal toxin ToxB from Pyrenophora tritici-repentis.

Transcriptional changes induced by Ptr ToxB

Water and ToxB-infiltrated leaves of sensitive cultivar Katepwa were collected at 0,3,9,14,24,48 hours post infiltration (hpi), total RNA was extracted and processed. Expression levels were obtained using Affymetrix GeneChip Wheat Genome Array


P-MTAB-25139

Plants were grown in a growth chamber under a cycle of 16 hours (h) of light at 22 C, and 8 h of darkness at 18 C

P-MTAB-25140

Native and heterologously (het) expressed ToxB was produced and purified as described (Figueroa Betts et al., 2011).

P-MTAB-25141

RNA for all replicate samples was isolated on the same day with the RNeasy Plant Mini Kit (Qiagen, Chatsworth, CA) following manufacturer's instructions. RNA concentration was measured with NanoDrop ND-1000 UV-Vis spectrophotometer (NanoDrop Technologies, Wilmington, DE) and resulted in 800-1200 ng/ul in 30 ul. 

P-MTAB-25142

Biotinylated cRNA was generated following the One-Cycle Target Labeling protocol (Affymetrix, Santa Clara, CA) from the GeneChip Expression Analysis Technical Manual (701021 Rev. 5). Biotinylated cRNA was synthesized from double-stranded cDNA using T7 RNA polymerase and a biotin-conjugated pseudouridine containing nucleotide mixture provided in the IVT Labeling Kit (Affymetrix).

P-MTAB-25143

Prior to hybridization, the cRNA was purified with GeneChip Sample Cleanup Modules (Affymetrix) and fragmented.Fluidic Station 450 was used to wash and stain the arrays with streptavidin-phycoerythrin (Molecular Probes, Eugene, OR), followed by biotinylated anti-streptavidin (Vector Laboratories, Burlingame, CA) according to standard antibody amplication protocol for eukaryotic targets.

P-MTAB-25144

Arrays were scanned with an Affymetrix GeneChip Scanner 3000 at 570nm

Transcription profiling by array of wheat leaves in response to the fungal toxin ToxB from Pyrenophora tritici-repentis

Submitter Supplied Information

Samples