Project description:After-ripening induced seed dormancy release in wheat is associated with mRNA oxidation. We used Affymetrix GeneChip Wheat Genome Array to identify mRNAs differentially oxidized by after-ripening Dormant and after-ripened seeds in dry state were used for isolating oxidized mRNAs and hybridization on Affymetrix GeneChip. After-ripened seeds were generated by storing dormant seeds at room temperature for 10 months.
Project description:affy_dormance_sunflower - affy_dormance_totalrna_sunflower - We focus our research on the release of embryonic dormancy in sunflower seeds and more precisely on the molecular mechanisms controlling seed dormancy release during dry after-ripening. Our data show an increase of ROS during dormancy release associated with an increase of stored mRNA oxidation during after-ripening as shown with determination of the RNA oxidation marker : 8-hydroxyguanosine (8OHG) To assess to role of mRNA oxidation in this process, we isolated 8OHG-containing mRNA to identify oxidized transcript using microarray analysis.-The samples compared are the following: 1) Population of 50 embryonic axis from dry dormant seed (D) ie. Seed which have been frozen at -30°C after harvest to keep dormancy 2) Population of 50 embryonic axis from dry non dormant ie. Seeds which have been after-ripened (stored) at 70% RH during 2 month. 3) Embryonic axis from dry dormant seed but after ripened at very low relative humidity (2% RH) (control of storage: S). Replicates correspond to different batches of seeds from the same harvest which have been treated (after ripened) independently. - We want to compare D vs ND; D vs S and ND vs S. 6 arrays - SUNFLOWER; dormant vs non dormant seed embryo
Project description:affy_dormance_sunflower - affy_dormance_totalrna_sunflower - We focus our research on the release of embryonic dormancy in sunflower seeds and more precisely on the molecular mechanisms controlling seed dormancy release during dry after-ripening. Our data show an increase of ROS during dormancy release associated with an increase of stored mRNA oxidation during after-ripening as shown with determination of the RNA oxidation marker : 8-hydroxyguanosine (8OHG) To assess to role of mRNA oxidation in this process, we isolated 8OHG-containing mRNA to identify oxidized transcript using microarray analysis.-The samples compared are the following: 1) Population of 50 embryonic axis from dry dormant seed (D) ie. Seed which have been frozen at -30°C after harvest to keep dormancy 2) Population of 50 embryonic axis from dry non dormant ie. Seeds which have been after-ripened (stored) at 70% RH during 2 month. 3) Embryonic axis from dry dormant seed but after ripened at very low relative humidity (2% RH) (control of storage: S). Replicates correspond to different batches of seeds from the same harvest which have been treated (after ripened) independently. - We want to compare D vs ND; D vs S and ND vs S.
Project description:affy_dormance_sunflower - affy_dormance_mrnaoxidized_sunflower - We focus our research on the release of embryonic dormancy in sunflower seeds and more precisely on the molecular mechanisms controlling seed dormancy release during dry after-ripening. Our data show an increase of ROS during dormancy release associated with an increase of stored mRNA oxidation during after-ripening as shown with determination of the RNA oxidation marker : 8-hydroxyguanosine (8OHG) To assess to role of mRNA oxidation in this process, we isolated 8OHG-containing mRNA to identify oxidized transcript using microarray analysis.-The samples compared are the following: 1) 8OHG containing mRNA isolated from total RNA extracted from a population of 100 embryonic axis of dry dormant seed (Dox) ie. Seed which have been frozen at -30°C after harvest to keep dormancy 2) 8OHG containing mRNA isolated from total RNA extracted from a population of 50 embryonic axis from dry non dormant ie. Seeds which have been after-ripened (stored) at 70% RH during 2 month (NDox) Replicates correspond to different batches of seeds from the same harvest, which have been treated (after ripened) independently and from which the 8OHG containing fraction have been isolated independently. 4 arrays - SUNFLOWER; dormancy
Project description:affy_dormance_sunflower - affy_dormance_mrnaoxidized_sunflower - We focus our research on the release of embryonic dormancy in sunflower seeds and more precisely on the molecular mechanisms controlling seed dormancy release during dry after-ripening. Our data show an increase of ROS during dormancy release associated with an increase of stored mRNA oxidation during after-ripening as shown with determination of the RNA oxidation marker : 8-hydroxyguanosine (8OHG) To assess to role of mRNA oxidation in this process, we isolated 8OHG-containing mRNA to identify oxidized transcript using microarray analysis.-The samples compared are the following: 1) 8OHG containing mRNA isolated from total RNA extracted from a population of 100 embryonic axis of dry dormant seed (Dox) ie. Seed which have been frozen at -30°C after harvest to keep dormancy 2) 8OHG containing mRNA isolated from total RNA extracted from a population of 50 embryonic axis from dry non dormant ie. Seeds which have been after-ripened (stored) at 70% RH during 2 month (NDox) Replicates correspond to different batches of seeds from the same harvest, which have been treated (after ripened) independently and from which the 8OHG containing fraction have been isolated independently.
Project description:Wheat seed dormancy is released by after-ripening, and germination and seminal root growth of after-ripened/non-dormant seeds can be inhibited by treatment with exogenous ABA. We used Affymetrix GeneChip Wheat Genome Array to detail transcriptional programs affected by after-ripening of dormant seeds and imbibition of after-ripened seeds in ABA.
Project description:To establish the basis for understanding molecular mechanism of seed germination response to temperature, we analyzed transcriptomes in freshly harvested dormant and dry stored after-ripened seeds. The after-ripened seeds started to show visible germination from 36h after the start of imbibition, and almost all the seeds germinated after 3 days. The freshly harvested seeds stayed dormant by imbibition at 26°C, and germination of the after-ripened seeds was almost completely inhibited at 34°C. Total RNA was prepared from 0 (dry), 6 and 24h imbibed seeds to find regulatory genes of seed dormancy and germination.
Project description:Wheat seed germination and seminal root growth can be inhibited by treatment with exogenous ABA We used Affymetrix GeneChip Wheat Genome Array to detail transcriptional programs affected by ABA during imbibition After-ripened seeds imbibed in ABA for 24 h were used for RNA extraction and hybridization on Affymetrix GeneChip. After-ripened seeds were generated by storing dormant seeds at room temperature for 10 months.
Project description:In present experiment we evaluated the effects of the utrasonication of winter wheat seeds on seedling growth and development. Effect of treatment on the gene transcription and DNA methylation of seedlings were evaluated.
Project description:Plant embryos can survive years in a desiccated, quiescent state within seeds. In many species, seeds are dormant and unable to germinate at maturity. They acquire the capacity to germinate through a period of dry storage called after-ripening (AR), a biological process that occurs at 5-15% moisture when most metabolic processes cease. Because stored transcripts will be among the first proteins translated upon water uptake, they likely impact germination potential. Transcriptome changes associated with the increased seed dormancy of the GA-insensitive <i>sly1-2</i> mutant, and with dormancy loss through <i>sly1-2</i> after-ripening or constitutive overexpression of the GA receptor (GID1b) were characterized in dry seeds. This experiment used the same seed batches as a previous experiment (E-MTAB-4782) to characterize transcriptional changes associated with the increased seed dormancy and dormancy loss in imbibing seeds. The <i>SLY1</i> gene encodes the F-box subunit of an SCF E3 ubiquitin ligase needed for GA-triggered proteolysis of DELLA repressors of seed germination. In the <i>sly1-2</i> mutant, GA-directed DELLA proteolysis cannot occur leading to DELLA protein accumulation and increased dormancy. <i>sly1-2</i> mutant seeds are fully dormant at 2 weeks of dry storage (0% germination), but germinate well with very long after-ripening (51% germination after 19 months). <i>sly1-2</i> seed germination can also be rescued by overexpression of the GA receptor, <i>GA-INSENSITIVE DWARF1b</i> (<i>GID1b-OE</i>), which resulted in 74% germination at 2 weeks of dry storage. In this experiment, we sampled dry seeds of wild-type L<i>er</i> at 2 weeks of dry storage (non-dormant), dormant <i>sly1-2</i> (2 weeks of dry storage; <i>sly1-2</i>(D)), long after-ripened <i>sly1-2</i> (non-dormant, 19 months of dry storage; <i>sly1-2</i>(AR)), and <i>sly1-2 GID1b-OE</i> (non-dormant, 2 weeks of dry storage). This experimental design allowed comparison between these transcriptomes in dry seeds to determine if dry seed stored mRNA differences contribute to the dormancy phenotypes observed once seeds are imbibed. Seeds for L<i>er</i> wt, <i>sly1-2</i>(D), and <i>sly1-2 GID1b-OE</i> were grown alongside each other under the same conditions and after-ripened for 2 weeks. Seeds from <i>sly1-2</i>(AR) were grown under the same conditions in advance of the other lines to allow for the long after-ripening requirement. RNA was extracted using a phenol-chloroform-based extraction from three biological replicates per treatment.