Transcriptome analysis of a red-flowered white clover mutant
Ontology highlight
ABSTRACT: Transcriptomes analysis of the petals from a red-flowered white clover mutant (red flowers) and its shade-treated counterpart (white flowers) grown under shaded conditions was performed using high-throughput sequencing. We obtained 121,626,564 and 130,577,944 clean reads in red-flowered mutant and treated counterpart, respectively. Of these clean reads, we respectively gained 125,350 and 99,638 unigene sequencces in two groups. As a result, a total of 157,964 unigenes were obtained with an average length of 728 bp and a median length of 1346 bp. These findings provideed insights into the expression profiles in red-flowered white clover mutant, and deepened our understanding of flower pigmentation in white clower.
Project description:Pink-flowered strawberry is a new promising ornamental flower derived from intergeneric hybridization (Fragaria × Potentilla) with bright color, prolonged flowering period and edible fruits. However, the transcriptional events underlying anthocyanins biosynthesis pathway have not been fully characterized in its petal coloration. The pigment compounds accumulated in its fruits were the same as cultivated strawberry, but different from in its flowers. To gain insights into the regulatory networks related to anthocyanin biosynthesis and identify key genes, we performed an integrated analyses of the transcriptome and metabolomes involved in red petals at three development stages (Bud stage (L), Coloration beginning stage (Z) and Big bud stage (D)) of pink-flowered strawberry. Transcript and metabolite profiles were generated through high-throughput RNA-sequencing and high-performance liquid chromatography coupled with mass spectrometry, respectively. The results showed that the main pigments of red and dark pink petals were anthocyanins, among which cyanidins were the main compounds. There were no anthocyanins detected in white-flowered hybrids. A total of 50 285 non-redundant unigenes were obtained from the transcriptome databases, among which 59 differentially expressed genes could be identified as putative homologues of flower coloration related genes. Based on a comprehensive analysis relating pigmentation compounds to gene expression profiles, the mechanism of flower color formation was examined in pink-flowered strawberry. Furthermore, a new hypothesis explaining the lack of color phenotype of the white-flowered strawberry hybrids from the level of the transcriptome. The expression patterns of FpDFR gene and FpANS gene corresponded to the accumulation patterns of cyanidin contents in pink-flowered strawberry hybrids with different shades of pink; Whereas other anthocyanin biosynthesis genes were weakly related flower color deepened. Moreover, FpANS, FpBZ1 and FpUGT75C1 genes were the key factors that lead to the inability to accumulate anthocyanins in the white petals of PFS hybrids. Meanwhile, the competitive effect of FpFLS gene and FpDFR gene may further inhibit anthocyanin synthesis. The data presented herein are important for understanding of the molecular mechanisms underlying the petal pigmentation and will be powerful for integrating into novel genes that are potential targets for breeding new valuable pink-flowered strawberry cultivars.
Project description:There are 16 organ samples (dry seeds, 24H imbibed seeds, 48H imbibed seeds, juvenile rosette, adult rosette, senescence leaves, cauline leaves, stems, young buds, mature flower buds, flowers, young siliques, mature siliques and old siliques) with triplicates. There are 17 samples of different environmental samples (0 h white, 1 h white, 6 h white, 24 h white, dark, blue, far-red and red lights, control, cold 2h, cold 6h, hot 2h, hot 6h, NaCl 2h, NaCl 6h, dry 2h and dry 6h) with triplicates.
Project description:Red clover (Trifolium pratense L.) is one of the most important legume forage species in temperate livestock agriculture. Tetraploid red clover cultivars are generally producing less seed than diploid cultivars. Improving the seed setting potential of tetraploid cultivars is necessary in order to utilize the high forage quality and environmentally sustainable nitrogen fixation of red clover. Two genotypes, one from cv.Tripo with weak seed setting and one from cv.Lasang with strong seed setting, were selected based on data from field experiments for transcriptome analysis of developing flower buds. De novo and reference based analyses of transcriptome assemblies were conducted to study the global transcriptome changes from early to late developmental stages of flower development of the two contrasting red clover genotypes. Transcript profiles, gene ontology enrichment and KEGG pathway analysis indicate that genes related to flower development, pollen pistil interactions, photosynthesis and embryo development are differentially expressed between the Tripo and Lasang genotypes. A significant number of genes related to pollination was overrepresented in Lasang, which might be a reason for its good seed setting ability. The candidate genes detected in this study might be used to develop molecular tools for breeding tetraploid red clover varieties with improved seed yield potentials.
Project description:Improvement of freezing tolerance of red clover (Trifolium pratense L.) would increase its persistence under cold climate. In this study, we assessed the freezing tolerance and compared the proteome composition of non-acclimated and cold-acclimated plants of two initial cultivars of red clover: Endure (E-TF0) and Christie (C-TF0) and of populations issued from these cultivars after three (TF3) and four (TF4) cycles of phenotypic recurrent selection for superior freezing tolerance. Through this approach, we wanted to identify proteins that are associated with the improvement of freezing tolerance in red clover. Recurrent selection performed indoor is an effective approach to improve the freezing tolerance of red clover. Significant improvement of freezing tolerance by recurrent selection was associated with differential accumulation of a small number of cold-regulated proteins that may play an important role in the determination of the level of freezing tolerance.
Project description:To investigate the mechanisms related to anti-ferroptotic effects of red clover extract, we performed differentially expressed genes analysis using data obtained from the RNA-seq of system xCT knockout mouse embryonic fibroblast cells.
Project description:To study the transcriptomic profile of wt and brc1 mutant axillary buds during the shade avoidance response, we simulated a canopy shade with a low R/FR light ratio. We treated plants with white light supplemented with far-red light (Red light = 29 μeinstein · m-2 seg-1, Far-Red light= 146 μeinstein · m-2 seg-1) for 8 hours. Control plants were left for 8 hours in white light (Red light = 29 μeinstein · m-2 seg-1, Far-Red light= 2.2 μeinstein · m-2 seg-1) .
Project description:White clover mosaic virus (WCMV) is a major pathogen of white clover (Trifolium repens L.), with significant effects on yield and persistence. Due to the absence of natural sources of WCMV resistance a transgenic strategy has been employed to produce plants constitutively expressing WCMV replicase gene derivatives, designed to inhibit the propagation of WCMV through an RNA silencing mechanism. A 12,000 feature oligonucleotide microarray has been used to identify global changes in host plant, in addition to virus genome-encoded gene expression associated with WCMV infection in non-transgenic and transgenic WCMV-resistant white clover. Pairwise comparison between the transcriptome of mock-inoculated non-transgenic and WCMV-inoculated transgenic plants provides clear evidence for substantial equivalence between these two genotype/treatments, and demonstrate the efficacy of the transgenic strategy. WCMV- inoculated non-transgenic plants exhibit elevated abundance of many virus-encoded, and host immune response-specific transcripts compared to the transgenic resistant plants or mock-inoculated non-transgenic plants. By contrast, relative to inoculated sensitive plants, the majority of significantly up-regulated genes in mock-inoculated non-transgenic plants or WCMV-inoculated transgenic plants are markers of healthy cellular function. These results, and the occurrence of levels of WCMV-encoded transcripts in inoculated transgenic plants equivalent to those in virus-free plants, confirm the validity of the transgenic RNA silencing approach.<br>