Transcriptomics

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Transcription profiling of wheat interacting with incompatible and compatible yellow rust in a Yr1-containing genotype


ABSTRACT: Transcriptional changes were monitored in the wheat cultivar Avocet*6/Yr1 following inoculation with avirulent and virulent wheat yellow rust (Puccinia striiformis f.sp. tritici) isolates using the Affymetrix wheat genome array GeneChip®. Seedlings of Avocet*6/Yr1 were grown to growth stage 12-13 (Zadoks et al., 1974) before inoculating with either the P.s. f.sp. tritici isolate 169E136 (Yr1-virulent) or 232E137 (Yr1-avirulent) or with talc for the Mock inoculation (Boyd and Minchin 2001). Plants were grown under a 16/8 hour photoperiod cycle, supplemented with sodium lighting (240 μmol m-2 s-1) at day/night temperatures of 20°C/15°C and a relative humidity of 60%, both before and after inoculation, in a spore-free, containment level 2 greenhouse. Leaf samples from six seedlings were collected 6, 12, 24, 48 and 72 hpi for RNA extraction and transcriptomics analysis from three independent biological experiments. Leaf tissue was ground under liquid nitrogen and total RNA extracted using the TRIzol reagent method, following the manufacturer’s instructions (Invitrogen, Carlsbad, CA). RNA from each time point (16 μg) were pooled to obtain 80 μg of total RNA for each treatment (mock inoculated control; avirulent isolate inoculated; virulent isolate inoculated). Pooled RNA samples were further purified using the Qiagen RNeasy mini-kit according to the manufacturer instructions (Qiagen) and RNA integrity was confirmed using the Agilent 2100 Bioanalyzer (Agilent). Affymetrix GeneChip processing, including RNA quality control, microarray hybridisation and data acquisition was performed through contract research services by the John Innes Genome Laboratory, John Innes Centre, Norwich, U.K. A total of nine hybridisations were performed. ****[PLEXdb(http://www.plexdb.org) has submitted this series at GEO on behalf of the original contributor, Osman Bozkurt. The equivalent experiment is TA25 at PLEXdb.]

ORGANISM(S): Triticum aestivum

PROVIDER: GSE31761 | GEO | 2011/08/31

SECONDARY ACCESSION(S): PRJNA145297

REPOSITORIES: GEO

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