Project description:The present study quantifies the transcriptomes of wild-type and transgenic Ubi::OsYHB rice seedlings (in the genetic background of Oryza sativa ssp. japonica CV Nipponbare) grown in the dark or under continous red light (Rc, at 50 µmol m-2 s-1) conditions. Overall design: WT (Nipponbare cultivar; Nip) and Ubi::OsYHB/Nip transgenic seedlings were grown at 28°C for 5 days in darkness or under continuous red light at 50 µmol m-2 s-1 (Rc50). Seedlings were harvested in subjective morning, immediately frozen in liquid nitrogen and strored at -80°C until RNA extraction. The expression of OsYHB is driven by the maize Ubiquitin promoter. Two biological replicates for each treatment. Two independent, genetically single-insertion, homozygous Ubi::OsYHB/Nip lines (#1 and # 9, with determined 41- and 23-fold overexpression levels of OsYHB in comparison to the wild-type control, respectively) were used. GeneChip 3' IVT Express Kit (Affymetrix) was used to synthesize and label aRNA.
Project description:The allene oxide synthase (AOS) branch and the hydroperoxide lyase (HPL) branch of the oxylipin pathway function in plant responses to diverse stresses and have potential cross-talks between each other in the biosynthesis and signaling regulation, but there is still an absence of direct evidence and detailed information about this communication. Here, we identified and characterized a jasmonates acid (JA) overproduction mutant, cea62, by screening the Constitutive Expression of AOS gene (cea) from rice T-DNA insertion mutant library. Map-based cloning was used to isolate the target gene as the hydroperoxide lyase OsHPL3 gene. The gene expression, HPL enzyme activity and its resulting products, (E)-2-hexenal, were all depleted in the cea62 mutant, which resulted in dramatic JA overproduction and activation of JA signaling in the mutant. Consistent with the formation of the lesion mimic phenotype and the timing of the induction for some defense responsive genes, the activation of JA biosynthesis and signaling was regulated in a developmental way, just as the way by which OsHPL3 was regulated in the wild type plant. Microarray data showed that the JA-governed defense response was greatly activated in the cea62 mutant plant and the cea62 plant obtained enhanced resistance to the bacterial blight pathogen Xanthomonasoryzaepvoryzae (Xoo) T1 strain. Wounding response was attenuated in the cea62 mutant at an early developmental stage, while it was partially recovered when JA levels were elevated at a later developmental stage in the cea62 mutant. But, the wounding response was not altered at different developmental stages in the wild type plant. These findings suggest that these two branches of the oxylipin pathways crosstalked in the biosynthesis and signaling pathways and cooperated with each other to function in diverse stress responses. We compared the gene expression profile in leaf tissues of the wild-type plant and the cea62 mutant after lesion mimic phenotype appeared two months after sowing. Total RNAs were extracted from leaf blades from the rice (Oryza sativa L.) wild-type Nipponbare plant and from leaf blades of the rice (Oryza sativa L.) cea62 mutant in the Nipponbare (ssp japonica) background at two months after sowing. Three replicates of the cea62 mutant and wild type were performed.
Project description:Oryza sativa Indica group IR29 (salt sensitive) seedlings were subjected to salt stress or control conditions and sampled at five time points over the course of 24 hours. RNA samples extracted were assayed using the Illumina HiSeq 2000 platform.