Project description:Purple-grain wheat are caused by anthocyanin accumulation in the seed coat. The anthocyanin biosynthesis and accumulation were affected by light in purple-grain wheat. The spikes of purple-grain wheat Luozhen No.1 were bagged with four-layer Kraft paper bags after pollination. To identify genes involved in the anthocyanin biosynthesis, we sequenced two pericarp cDNA libraries, D20 (20 DAP) of shading treatment, and L20 (20 DAP) of untreated control using an Illumina HiSeqTM 2000.
Project description:Purple-grain wheat are caused by anthocyanin accumulation in the seed coat. But little is known about molecular mechanism of anthocyanin biosynthesis. The anthocyanin biosynthesis and accumulation were affected by light in purple-grain wheat. The spikes of purple-grain wheat Luozhen No.1 were bagged with four-layer Kraft paper bags after pollination. To identify genes involved in the anthocyanin biosynthesis, we sequenced four pericarp cDNA libraries, D15 (15 DAP), D20 (20 DAP) of shading treatment, and L15 (15 DAP), L20 (20 DAP) of untreated control using an Illumina HiSeqTM 2000. After quality control, raw reads are filtered into clean reads which will be aligned to the reference sequences. The alignment data is utilized to calculate distribution of reads on reference genes and mapping ratio, and proceed with downstream analysis including gene and isoform expression, deep analysis based on gene expression (PCA/correlation/screening differentially expressed genes and so on),exon expression, gene structure refinement, alternative splicing, novel transcript prediction and annotation, SNP detection, Indel detection. Further, we also perform deep analysis based on different expression genes, including Gene Ontology (GO) enrichment analysis, Pathway enrichment analysis, cluster analysis, and finding transcriptor factor.
Project description:Purpose: The present study aimed to investigate the anthocyanin components and identify relevant regulatory genes in purple wheat grain by carrying out transcriptome analyses. Methods: The seeds of purple grain wheat and white grain wheat were collected 30 days after flowering, and three biological replicates were set. Total RNA was isolated and purified using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) following the manufacturer's procedure. The RNA amount and purity of each sample was quantified using NanoDrop ND-1000. Then synthesizing the fragmented RNA into cDNA through the action of reverse transcriptase, and finally obtaining acDNA library. At last, we performed the 2×150bp paired-end sequencing (PE150) on an Illumina Novaseq™ 6000 following the vendor's recommended protocol. Results: A total of 10440 diferentially expressed genes were signifcantly enriched by RNA sequencing. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed signifcantly enriched flavonoid biosynthesis and anthocyanin biosynthesis in CW_S versus W_S. And the ANS and UFGT genes were predicted as core genes in anthocyanin biosynthesis. Conclusions: Our study represents the detailed analysis of wheat grain transcriptomes, with biologic replicates, generated by RNA-seq technology. Through this study, we speculated that ANS and UFGT genes are the core genes of anthocyanin biosynthesis.The significant differences of these genes affect the synthesis of anthocyanins in wheat grains, and thus affect the grain color of wheat.
Project description:Colored-leaf plants are increasingly popular and have been attracting more and more attentions. However, studies about the anthocyanin components and molecular mechanisms of anthocyanin biosynthesis in QHP and ZSY was still unclear. In present study, an integrated metabolite and transcriptome profiling analysis was performed to explore the anthocyanin compositions and the specific regulatory network of anthocyanin biosynthesis in purple leaves of QHP and ZSY. A total of 39 anthocyanin compounds were detected based on the LC-MS/MS analysis. Of these, 12 cyanidins, 7 pelargonidins, 5 delphindins, and 5 procyanidins are the major anthocyanin compounds, which were significantly differentially accumulated in purple leaves of QHP and ZSY. The major genes associated with anthocyanin biosynthesis, including structural genes and TFs, were differentially expressed in the purple leaves of QHP and ZSY through RNA-seq data analysis, some of which were further assessed by qRT-PCR. Correlation between RNA-seq analysis and metabolite profiling showed that the expression pattern of some differentially expressed genes in anthocyanin biosynthes pathway were closely correlated with the differential accumulation of anthocyanins. In addition, one member of SG5 subgroup of R2R3-MYB TFs, Podel.04G021100, shows a similar expression pattern to some structure genes and closely correlates with sixteen anthocyanin compounds, indicating that the MBY TF (Podel.04G021100) may be involved in the regulation of anthocyanin biosynthesis. The above results not only make us systematic and comprehensive understand the anthocyanin components and corresponding molecular mechanisms of anthocyanin biosynthesis in purple leaves of QHP and ZSY, but also contribute to the promotion and application of anthocyanins in colored-leaf poplars.
Project description:Wheat yield is correlated significantly with grain size which is established during morphological stage. In present study, morphological stage of developing wheat grain were analyzed by RNA-seq.This research will help us to understand the mechnism underlying of grain development. This is the first study on gene expression profiling of morphological stage of developing wheat grain and the results may aid the identification of pathways and genes associated with seed development in wheat.
Project description:Transcriptional comparison of developing grains between two wheat genotypes with contrasting levels of minerals in grain, using Affymetrix GeneChip® Wheat Genome Array.
Project description:Oilseed rape is both an important oleaginous crop and agriculture sightseeing crop whereas has relatively scanty flower color. As natural flavonoids, Anthocyanin are responsible for the attractive red, purple, and blue colors of various tissues in higher plants, especially for the ornamental plants flower. One Brassica napus-Orychophragmus violaceus disomic addition line (M4) obtained previously exhibits red petals whichresult from anthocyanin biosynthesis. Transcriptome analysis of M4, B. napus (H3), natural individuals of O. violaceus with purple petals (OvP) and white petals (OvW) revealed that most of structural genes for the anthocyanin synthesis were up-regulated in both M4 and OvP, especially key gene ANS in the last step. Reads assembling and sequence alignment showed that the regulatory DEG PAP2 in M4 was from the transcript of O. violaceus. OvPAP2 was transformed into Arabidopsis thaliana and B. napus driven by the CaMV35S promoter and the rape petal-specific prompter XY355. Transgenic A. thaliana plants showed different levels of purple pigments in most of the organs, including the petals, and transgenic B. napus flowers exhibited restricted accumulation of anthocyanins in stamens when driven by CaMV35S promoter, but generated both red petals and anthers driven by the XY355 promoter. These results provided a platform for expounding the anthocyanin biosynthesis pathway in B. napus petals and give a successful case for flower color modification of the agriculture sightseeing rape.
Project description:Transcriptional comparison of developing grains between two wheat genotypes with contrasting levels of minerals in grain, using Affymetrix GeneChipM-BM-. Wheat Genome Array. Gene expression data of two wheat genotypes with high and low grain mineral concentration at two seed development stages (14, and 28 days after anthesis)
Project description:Allohexaploid bread wheat (Triticum aestivum, L.) provides ~ 20% of calories consumed by humans. Hitherto lack of genome sequence for the three homoelogous and highly similar bread wheat genomes (A, B and, D) impeded expression analysis of the grain transcriptome. We used novel genome information to analyze the cell type specific expression of homeologous genes in the developing wheat grain.
Project description:Development of wheat (Triticum aestivum L.) grain mainly depends on the processes of starch synthesis and storage protein accumulation, which are critical for grain yield and quality. However, the regulatory network underlying the transcriptional and physiological changes of grain development is still not clear. Here, we combined ATAC-seq and RNA-seq to discover the chromatin accessibility and gene expression dynamics during these processes. We found that the chromatin accessibility changes are tightly associated with differential expressions and the proportion of distal ACRs were increased gradually during grain development. Specific transcription factor (TF) binding sites were enriched at different stages, and were diversified among the 3 subgenomes. We further predicted the potential interactions between key TFs and genes related with starch and storage protein biosynthesis and found different copies of some key TFs played diversified roles. Overall, our findings have provided numerous resources and illustrated the regulatory network during wheat grain development, which would shed lights on the improvement of wheat yields and qualities.