Project description:Here, we performed the cleavage under targets and tagmentation (CUT&Tag) assay, a recently developed technology for protein-DNA interaction, in protoplasts extracted from wheat root tissues to genome-widely identity the potential targets of TaSPL6-D. Using CUT-Tag-seq, we obtained 7,403 confidential peaks from two biological replicates, with which 2688 genes was associated. Zooming in the promoter region, GTAC motif was significantly enriched in the peaks. Gene Ontology analysis of genes with binding peaks in the promoter region revealed that, the most enriched gene term is ‘response to temperature/heat stimuli’, which is consistent with previous studies on the functions of TaSPL6-D orthologs in rice and Arabidopsis. The following enriched terms are genes of ‘regulation of response to salt stress’, ‘regulation of response to osmotic stress’, and particularly ‘positive regulation of response to salt stress’, implying TaSPL6-D is an upstream regulator of salt stress response.
Project description:We aimed to identify targets of miRNAs during wheat grain development by using degradome sequencing approach. Two degradome libraries were constructed from wheat grains.
Project description:We aimed to identify targets of miRNAs during wheat grain development by using degradome sequencing approach. Two degradome libraries were constructed from wheat grains. Verification of miRNA targets from two degradome libraries in developing wheat grains.
Project description:Water-deficit stress negatively affects wheat yield and quality. Abiotic stress on the parental plants during reproduction could have transgenerational effects on the progenies. Here we investigated the transgenerational influence of pre-anthesis water-deficit stress by detailed analysis of the yield components, grain quality traits, and physiological traits in durum wheat. Next-generation sequencing analysis profiled the small RNA-omics, mRNA transcriptomics, and mRNA degradomics in the progenies. Parental water-deficit stress had positive impacts on the progenies in certain traits like harvest index and protein content in given genotype. Small RNA-seq identified 1739 conserved and 774 novel microRNAs (miRNAs). Transcriptome-seq characterised the expression of 66,559 genes while degradome-seq profiled the miRNA-guided mRNA cleavage dynamics. Differentially expressed miRNAs and genes were identified, with significant regulatory patterns subject to trans- and inter- generational stress. Integrated analysis based on the three omics revealed the significant biological interactions between stress-responsive miRNA and targets, with possible contributions towards transgenerational stress tolerance via pathways such as hormone signalling and nutrient metabolism. Our study provides the first confirmation of the transgenerational effects of water-deficit stress in durum wheat. New insights gained on the molecular level indicate that key miRNA-mRNA modules are potential candidates in transgenerational stress improvement.
Project description:The goals of this study were to compare the transcriptome of six genotypes of wheat grown under the normal conditions by RNA-Seq and to study the root architecture in drought sensitive and tolerant genotypes.
Project description:To explore the effect of stable RNAi on the small RNA (sRNA) population in wheat, we constructed a sRNA library from hexaploid wheat that expresses an RNAi construct under the 35S promoter that targets the endogenous NO APICAL MERISTEM (TaNAM) gene. The presence of this RNAi transgene causes a 40% reduction in expression of the target genes as measured by quantitative RT-PCR and significantly delays senescence and reduces remobilization of N, Fe, and Zn to the grain.
Project description:Background: MicroRNAs regulate various biological processes in plants. Considerable data are available on miRNAs involved in the development of rice, maize and barley. In contrast, little is known about miRNAs and their functions in the development of wheat. In this study, five small RNA (sRNA) libraries from wheat seedlings, flag leaves, and developing seeds were developed and sequenced to identify miRNAs and understand their functions in wheat development. Results: Twenty-four known miRNAs belonging to 15 miRNA families were identified from 18 MIRNA loci in wheat in the present study, including 15 (9 MIRNA loci) first identified in wheat, 13 miRNA families (16 MIRNA loci) being highly conserved and 2 (2 MIRNAs loci) moderately conserved. In addition, fifty-five novel miRNAs were also identified. The potential target genes for 15 known miRNAs and 37 novel miRNAs were predicted using strict criteria, and these target genes are involved in a wide range of biological functions. Four of the 15 known miRNA families and 22 of the 55 novel miRNAs were preferentially expressed in the developing seeds with logarithm of the fold change of 1.0~7.6, and half of them were seed-specific, suggesting that they participate in regulating wheat seed development and metabolism. From 5 days post-anthesis to 20 days post-anthesis, miR164 and miR160 increased in abundance in developing seeds, whereas miR169 decreased, suggesting their coordinating functions in the different developmental stages of wheat seed. Moreover, eight known miRNA families and 28 novel miRNAs exhibited tissue-biased expression in wheat flag leaves, with the logarithm of the fold changes of 0.5~5.2. The putative targets of these tissue-preferential miRNAs were involved in various metabolism and biological processes, suggesting complexity of the regulatory networks in different tissues. Our data also suggested that wheat flag leaves have more complicated regulatory networks of miRNAs than developing seeds. Conclusions: Our work identified and characterised wheat miRNAs, their targets and expression patterns. This study is the first to elucidate the regulatory networks of miRNAs involved in wheat flag leaves and developing seeds, and provided a foundation for future studies on specific functions of these miRNAs.
Project description:Investigation of genome wide expression level changes during 11 stages of wheat grain development in normal growth conditions (19°C).