Project description:Transcript changes in response to low temperature Total RNA for RNA-seq analysis were extracted from wheat leaf tissues with three biological replicates for each growth condition.
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare NGS-derived Triticum aestivum transcriptome (RNA-seq) profiling methods and to evaluate genotypes associated with resistance against the Wheat dwarf virus. Methods: Triticum aestivum mRNA profiles of genotypes associated with resistance against the Wheat dwarf virus were generated by deep sequencing, in four replicates, using Illumina. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays. Conclusions: Our study represents the first detailed analysis of Triticum aestivum transcriptomes, with biologic replicates, generated by RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of expression profiles. Our results show that NGS offers a comprehensive and more accurate quantitative and qualitative evaluation of mRNA and miRNA content within a cell or tissue. We conclude that RNA-seq based transcriptome characterization would expedite genetic network analyses and permit the dissection of complex biologic functions.
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:The aim of this study was to identify genetic mechanisms of tolerance to active de-acclimation in winter wheat by comparing four tolerant and four susceptible accessions (cultivars and advanced breeding lines). The plants were kept in controlled conditions. 21 days after sowing, the plants were cold acclimated (3 weeks, 4/2 °C, day/night temperature; photoperiod of 10/14 h). Cold acclimation was followed by 7 days of de-acclimating conditions, mimicking a mid-winter warm spell: 12/5°C, day/night temperature; photoperiod of 10/14 h). The fragments of leaves were sampled in three biological replicates (three different plants of the same accession) in three timepoints: just before cold-acclimation (CA-0), after cold-acclimation (CA-21), and after de-acclimation (DA-28). Total RNA samples were sequenced on Illumina NovaSeq 6000 platform in PE150 bp mode. Differences in expression for a given accession were tested between all time-points, averaging the results for three biological replicates. It has been confirmed that active de-acclimation is not simply the inverse of cold acclimation, and that susceptible forms show significantly more changes in expression as a result of de-acclimation than tolerant forms. This provides further support for the hypothesis that tolerance to de-acclimation consists mainly of the absence or weakest response to temperature rise. Among the genes whose expression was significantly altered as a result of de-acclimation, a significant group were genes related to the defence response to stress. In wheat particularly plant hormone related genes, and plant hormones, seem to be playing a crucial role in the response to active de-acclimation, especially ABA and SA. Our results also suggest that in wheat the period of cold-acclimation might be decisive in the later active de-acclimation tolerance.