Project description:MicroRNAs (miRNAs) are small non-coding RNAs that play important roles by regulating other genes. Rose, Rosa rugosa Thunb., is an important ornamental and edible plant, yet there are only a few studies on the miRNAs and their functions in rose. Here we carried out computational and experimental analysis of miRNAs, phased small interfering RNAs (phasiRNAs) and mRNAs in rose by analyzing 10 small RNA sequencing profiles from roots, petals, pollens, stamens, and leaves and 4 RNA-seq profiles in leaves and petals of rose. To identify the targets of miRNAs and phasiRNAs, we produced a degradome profile for rose leaf which is analyzed using the SeqTar algorithm. This study identified 25 conserved pre-miRNAs, of which 24 have not been reported previously. We also found 22 novel pre-miRNAs. Three hundred and thirty nine 21 nucleotide (nt) PHAS loci, and forty nine 24 nt PHAS loci were also identified. We identified more than 19,000 putative targets of the conserved miRNAs/tasiRNAs using a criteria of less than 4 mismatches between miRNA and targets. Among these targets, 592 have shown significant accumulation of degradome reads. Our results demonstrate that the miR482 family triggers the generations of phasiRNAs by targeting nucleotide-binding, leucine-rich repeat (NB-LRR) disease resistance genes in rose. Our results also suggest that the deregulated genes in leaves and petals are significantly enriched in GO and KEGG pathways related to metabolic processes and photosynthesis. These results significantly enhanced our knowledge of the miRNAs and phasiRNAs, as well as their potential functions in rose.
Project description: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.
Project description:MicroRNAs (miRNAs) are small non-coding RNAs that regulate target mRNAs by inducing degradation or preventing translation of their target mRNAs. Rose, Rosa rugosa Thunb., is an important ornamental and edible plant, yet there are only a few studies on the miRNAs of rose. Here we carried out computational and experimental analysis of miRNAs and phased small interfering RNAs (phasiRNAs) in rose by analyzing 10 small RNA profiles from roots, petals, pollens, stamens, and leaves. To identify the targets of miRNAs and phasiRNAs, we generated a degradome profile for rose leaf which is analyzed using the SeqTar algorithm. This study identified 25 conserved pre-miRNAs, of which 24 have not been reported previously. We also found 22 novel pre-miRNAs. Three hundred and thirty nine 21 nucleotide (nt) PHAS loci, and forty nine 24 nt PHAS loci were also identified. We identified more than 4000 putative targets of the conserved miRNAs using a criteria of less than 4 mismatches between miRNA and targets. Among these targets, at least 171 have shown significant accumulation of degradome reads. Our results demonstrate that the miR482 family triggers the generations of phasiRNAs by targeting nucleotide-binding, leucine-rich repeat (NB-LRR) disease resistance genes in rose. These results significantly enhanced our knowledge of the miRNAs and phasiRNAs, as well as their potential functions in rose.
Project description:MicroRNAs (miRNAs) are small non-coding RNAs that regulate target mRNAs by inducing degradation or preventing translation of their target mRNAs. Rose, Rosa rugosa Thunb., is an important ornamental and edible plant, yet there are only a few studies on the miRNAs of rose. Here we carried out computational and experimental analysis of miRNAs and phased small interfering RNAs (phasiRNAs) in rose by analyzing 10 small RNA profiles from roots, petals, pollens, stamens, and leaves. To identify the targets of miRNAs and phasiRNAs, we generated a degradome profile for rose leaf which is analyzed using the SeqTar algorithm. This study identified 25 conserved pre-miRNAs, of which 24 have not been reported previously. We also found 22 novel pre-miRNAs. Three hundred and thirty nine 21 nucleotide (nt) PHAS loci, and forty nine 24 nt PHAS loci were also identified. We identified more than 4000 putative targets of the conserved miRNAs using a criteria of less than 4 mismatches between miRNA and targets. Among these targets, at least 171 have shown significant accumulation of degradome reads. Our results demonstrate that the miR482 family triggers the generations of phasiRNAs by targeting nucleotide-binding, leucine-rich repeat (NB-LRR) disease resistance genes in rose. These results significantly enhanced our knowledge of the miRNAs and phasiRNAs, as well as their potential functions in rose.
Project description:affy_duplicature_lyon_rose. The objective is to identify the genes involved in petal doubling in rose. In this study we are using two rosa gallica genotypes: wild-type (simple flower rose) and Cardinal de Richelieu (double flower rose), and two rosa hybrida genotypes : Souvenir de la Malmaison, which has about 110 petal, and its bud sport cultivar, Souvenir de St Anne’s. In this study, we used a microarray approach to compare the transcriptome of double flower rose (CDR) versus simple flower rose (G). The objective is to identify genes whose expression is associated with the double flower phenotype. These genes are putative candidates involved in the control of petal organ number per flower. Floral buds were dissected under a microscope and pooled in eppendorf tubes. Tissue samples were harvested at the same time during 3 weeks in April 2007. Total RNA was extracted from the pools of flowers using the Plant RNA kit (Macherey Nagel), and then used to hybridize Rosa-Affymetrix microarrays. Keywords: genotype comparison
Project description:affy_duplicature_lyon_rose. The objective is to identify the genes involved in petal doubling in rose. In this study we are using two rosa gallica genotypes: wild-type (simple flower rose) and Cardinal de Richelieu (double flower rose), and two rosa hybrida genotypes : Souvenir de la Malmaison, which has about 110 petal, and its bud sport cultivar, Souvenir de St Anne’s. In this study, we used a microarray approach to compare the transcriptome of double flower rose (CDR) versus simple flower rose (G). The objective is to identify genes whose expression is associated with the double flower phenotype. These genes are putative candidates involved in the control of petal organ number per flower. Floral buds were dissected under a microscope and pooled in eppendorf tubes. Tissue samples were harvested at the same time during 3 weeks in April 2007. Total RNA was extracted from the pools of flowers using the Plant RNA kit (Macherey Nagel), and then used to hybridize Rosa-Affymetrix microarrays. Keywords: genotype comparison 8 arrays - rose. 4 genotypes, 2 replicates each.