概要: Rosa chinensis ‘Pallida’ (Rosa L.) is one of the most important ancient rose cultivars originating from China. It contributed the ‘tea scent’ trait to modern roses. However, little information is available on the gene regulatory networks involved in scent biosynthesis and metabolism in Rosa. In this study, the transcriptome of R. chinensis ‘Pallida’ petals at different developmental stages, from flower buds to senescent flowers, was investigated using Illumina sequencing technology. De novo assembly generated 89,614 clusters with an average length of 428 bp. Based on sequence similarity search with known proteins, 62.9% of total clusters were annotated. Out of these annotated transcripts, 25,705 and 37,159 sequences were assigned to gene ontology and clusters of orthologous groups, respectively. The dataset provides information on transcripts putatively associated with known scent metabolic pathways. Digital gene expression (DGE) was obtained using RNA samples from flower bud, open flower and senescent flower stages. Comparative DGE and quantitative real time PCR permitted the identification of five transcripts encoding proteins putatively associated with scent biosynthesis in roses. The study provides a foundation for scent-related genes discovery in roses.
项目整体设计: Examination of transcriptome of Rosa chinensis 'Pallida' by RNA-seq.
All of Rosa chinensis 'Pallida' used in this study were grown under controlled conditions in a greenhouse of Flower Research Intitute of Yunnan Academy of Agricultural Sciences.
处理方案:
No treatment
提取方案:
Total RNAs were extracted from Flower bud, open flower and senescent flower with CTAB reagent method (Invitrogen).
建库方案:
Equal volumes of RNA from flower buds, open flowers and senescent flowers were pooled. RNA quality and quantity were verified using a NanoDrop 1000 spectrophotometer and an Agilent 2100 Bioanalyzer prior to further processing. Total RNA was treated with DNase I prior to library construction, and poly-(A) mRNA was purified with Magnetic Oligo (dT) Beads. Double-stranded cDNA was further subjected to end-repair using T4 DNA polymerase, the Klenow fragment, and T4 polynucleotide kinase followed by a single A dNTP base addition using Klenow 3’ to 5’ exo-polymerase, then ligated with an adapter or index adapter using T4 DNA ligase.
