Project description:Petunia is an excellent model system, especially for genetic, physiological and molecular studies. Thus far, however, genome-wide expression analysis has been rarely applied because of the lack of sequence information. We applied next-generation sequencing to generate, through de novo read assembly, a large catalogue of transcripts for Petunia axillaris and Petunia inflata. On the basis of the transcriptome of each species, comprehensive microarray chips for gene expression analysis were established and used for the analysis of global- and organ-specific gene expression in both species. In addition, microarray analysis was applied to explore the molecular basis of the seed coat defects in Petunia hybrida mutants, homozygous for a null allele of the AN11 gene, encoding a WDR transcription regulator. Among the transcripts differentially expressed in an11 seeds compared to wild type, many expected targets of AN11 were found but also several interesting new candidates that might play a role in morphogenesis of the seed coat. Our results validate the combination of next-generation sequencing with microarray analyses strategies to identify the transcriptome of two petunia species without previous knowledge of their genome, and to develop comprehensive chips as useful tools for the analysis of gene expression in P. axillaris, P. inflata and P. hybrida.
Project description:Petunia is an excellent model system, especially for genetic, physiological and molecular studies. Thus far, however, genome-wide expression analysis has been rarely applied because of the lack of sequence information. We applied next-generation sequencing to generate, through de novo read assembly, a large catalogue of transcripts for Petunia axillaris and Petunia inflata. On the basis of the transcriptome of each species, comprehensive microarray chips for gene expression analysis were established and used for the analysis of global- and organ-specific gene expression in both species. In addition, microarray analysis was applied to explore the molecular basis of the seed coat defects in Petunia hybrida mutants, homozygous for a null allele of the AN11 gene, encoding a WDR transcription regulator. Among the transcripts differentially expressed in an11 seeds compared to wild type, many expected targets of AN11 were found but also several interesting new candidates that might play a role in morphogenesis of the seed coat. Our results validate the combination of next-generation sequencing with microarray analyses strategies to identify the transcriptome of two petunia species without previous knowledge of their genome, and to develop comprehensive chips as useful tools for the analysis of gene expression in P. axillaris, P. inflata and P. hybrida. The manuscript describes the creation by next generation sequencing of a large catalogue of the transcriptome of the two Petunia species, that are considered to represent the natural material from which the breeders selected their varieties. This submission represents the transcriptome component of study. The high throughput sequencing data were submitted to SRA (accession numbers: SRA027293, SRP004866.1, SRX036999.2, SRX036998.2).
Project description:Purpose: The goal of this study are to reveal the internal mechanism of Bacillus cereus G2 increased Glycyrrhiza uralensis Fisch. seedlings salt-tolerance by RNA-Seq. Methods: mRNA profiles of Glycyrrhiza uralensis Fisch. Seedling in four treatment: control treatment, G2 treatment, salt treatment, salt and G2 treatment. Results: We mapped about 3 million sequence reads per sample to the G. uralensis transcriptome. A total of 35,831 genes in all samples of G. uralensis were identified and quantified by transcriptions, among which 3608 DEGs were identified. There are 1589, 623, 469 and 927 DEGs in S vs CK, CK+B vs CK, S+B vs S and S+B vs CK+B comparisons, respectively. Validation of expression levels for 12 randomly selected DEG candidates was carried out by quantitative real-time PCR (qRT-PCR). The results showed high congruence between RNA-Seq and qRT-PCR results (coefficient of determination R2 =0.9088) indicating the reliability of RNA-Seq quantification of gene expression. Conclusions: Our study help to better understand the underlying molecular mechanisms of G2 improve the salt tolerance of G. uralensis.