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The global wheat gene expressional responses to fusaoctaxin A

ABSTRACT: Purpose: To assess global changes of wheat coleoptiles gene expression in response to fusaoctaxin A which is a linear, C-terminally-reduced and D-amino acid residue-rich octapeptide, as the product of the two non-ribosomal peptide synthetases encoded by Fusarium graminearum fg3_54 cluster. Methods: For RNA seq of wheat coleoptiles, three biological replicates of mock-inoculated coleoptiles, wild-type PH-1 inoculated coleoptiles (3 dpi), ∆fg3_54 inoculated coleoptiles (3 dpi), after 3 hours fusaoctaxin A-treated coleoptiles, after 3 hours control (0.5% Tween-20) treated coleoptiles, after 24 hours fusaoctaxin A-treated coleoptiles and after 24 hours control (0.5% Tween-20) treated coleoptiles were sequenced. Results: For each sample, 33-95 M RNA-seq clean reads were obtained mapped to the Triticum aestivum Chinesespring genome sequence.Our results of RNA sequencing indicated fusaoctaxin does not induce PR gene expression, but suppress plant immunity and chloroplastc activity. Conclusions: Fusaoctaxin A represents an unprecedented virulence factor which can manipulate chloroplast and suppress host immunity.

ORGANISM(S): Triticum aestivum

PROVIDER: GSE117934 | GEO | 2019/01/09

REPOSITORIES: GEO

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Project description:Transcriptional changes were monitored in the wheat cultivar Renan 24 hours post i noculation with adapted and non-adapted Magnaporthe isolates using the Affymetrix wheat genome array GeneChip®. Wheat plants cv. Renan were grown in a peat and sand (1:1) mix at 23 C in a Sanyo Fitotron growth cabinet (Sanyo Gallenkamp PLC, Loughborough, U.K.) with a 16/8 h, light/dark cycle. Three Magnaporthe isolates were used in this expt, two wheat-adapted isolates (BR32, BR37) and one wheat non-adapted isolate (BR29). Magnaporthe isolates were grown for eleven days on Complete Media Agar at 25 C under a 16/8h, light/dark cycle. Conidia were harvested by flooding the plates with 5 mL of sterile inoculation solution [0.25% (w/v) gelatine and 0.01% (v/v) Tween 20] and scraping the conidia from the surface using a sterile glass rod. Conidia were filtered through sterile miracloth and the density adjusted to 1 x 10 5 conidia mL-1 with inoculation solution. Fourteen day old wheat seedlings mist inoculated with 4 mL of a Magnaporthe conidia suspension and plants were sealed in plastic propagators to maintain relative humidity c.100% and kept at 25 C in the dark for the first 24 hours post inoculation (hpi). Inoculation solution without Magnaporthe conidia was used as a mock-inoculation control. Leaf samples were collected 24 hpi for transcriptomics analysis from three independent biological experiments. Leaf tissue was ground under liquid nitrogen and total RNA extracted using a QIAquick RNeasy Plant Extraction Kit (Qiagen, Hilden, Germany), followed by TURBO DNaseTM (Ambion, Texas, U.S.A.) treatment. RNeasy Mini Spin column purification (Qiagen) was used to further purify RNA samples for array hybridisation. RNA quality checks, cRNA conversion and Affymetrix genome array hybridisation was carried out by the Nottingham Arabidopsis Stock Centre (NASC) array hybridisation service (http://affymetrix.arabidopsis.info/). ****[PLEXdb(http://www.plexdb.org) has submitted this series at GEO on behalf of the original contributor, Graham McGrann. The equivalent experiment is TA24 at PLEXdb.] pathogen isolates: Mock-inoculated (Control)(3-replications); pathogen isolates: Wheat non-adapted Magnaporthe isolate BR29(3-replications); pathogen isolates: Wheat adapted Magnaporthe isolate BR32(3-replications); pathogen isolates: Wheat adapted Magnaporthe isolate BR37(3-replications)

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The global wheat gene expressional responses to fusaoctaxin A

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