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Revealing impaired pathways in regulatory mutant by high-throughput characterization of Petunia axillaris and Petunia inflata transcriptomes.

ABSTRACT: 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.

ORGANISM(S): Petunia x hybrida Petunia axillaris Petunia integrifolia subsp. inflata

PROVIDER: GSE28120 | GEO | 2012/02/29

SECONDARY ACCESSION(S): PRJNA139839

REPOSITORIES: GEO

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Revealing impaired pathways in regulatory mutant by high-throughput characterization of Petunia axillaris and Petunia inflata transcriptomes.

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).

2012-02-29 | E-GEOD-28120 | biostudies-arrayexpress

Petunia hybrida

Project description:Petunia hybrida Transcriptome

| PRJNA572578 | ENA

Comprehensive transcriptional profiling analysis reveals candidate regulators in cold-stressed seedlings of Petunia hybrida

Project description:Investigation of gene expression level changes in Petunia hybrida seedlings subjected to cold at 2°C for 0.5 h, 2 h, 24 h and 5 d, compared to the CK.

2015-01-15 | GSE64963 | GEO

Petunia x hybrida

Project description:Petunia x hybrida overview

| PRJNA158195 | ENA

High-throughput sequencing of small RNAs in Petunia

Project description:Small RNAs (21-24 nt) are pivotal regulators of gene expression that guide both transcriptional and post-transcriptional silencing mechanisms in diverse eukaryotes, including most if not all plants. MicroRNAs (miRNAs) and short interfering RNAs (siRNAs) are the two major types, both of which have a demonstrated and important role in plant development, stress responses and pathogen resistance. In this work, we used a deep sequencing approach (Sequencing-By-Synthesis, or SBS) to develop sequence resources of small RNAs from Petunia x hybrida tissues (including leaves and flowers). The high depth of the resulting datasets enabled us to examine in detail critical small RNA features as size distribution, tissue-specific regulation and sequence conservation between different organs in this species. We also developed database resources and a dedicated website (http://smallrna.udel.edu/) with computational tools for allowing other users to identify new miRNAs or siRNAs involved in specific regulatory pathways, verify the degree of conservation of these sequences in other plant species and map small RNAs on genes or larger regions of the genome under study. Small RNA libraries were derived from leaves and flowers (see Series GSE13764) of Petunia x hybrida. Total RNA was isolated using the Plant RNA Purification Reagent (Invitrogen), and submitted to Illumina (Hayward, CA, http://www.illumina.com) for small RNA library construction using approaches described in (Lu et al., 2007) with minor modifications. The small RNA libraries were sequenced with the Sequencing-By-Synthesis (SBS) technology by Illumina. PERL scripts were designed to remove the adapter sequences and determine the abundance of each distinct small RNA. We thank Ana Dorantes-Acosta and Richard A. Jorgensen for providing the plant material as well as Kan Nobuta and Gayathri Mahalingam for assistance with the computational methods.

2014-04-24 | E-GEOD-57052 | biostudies-arrayexpress

N1-methyladenosine methylome in messenger RNA in petunia

Project description:N1-methyladenosine is a unique base methylation because it blocks Watson-Crick base paring and introduces a positive charge. Previous studies showed that m1A is prevalent in yeast and mammals mRNA and has a functional role in promoting translation of methylated mRNA. However, little is known about its abundance, topology and dynamics in plant mRNA. In this study, dot blotting and LC–MS/MS analyses reveal a dynamic pattern of m1A mRNA modification in various tissues and at different developmental stages in petunia (Petunia hybrida). Transcriptome-wide profiling of mRNA m1A in petunia was reported by applying m1A mRNA immunoprecipitation followed by a deep-sequencing approach (m1A-seq). m1A-seq analysis identified 4993 m1A peaks in 3231 expressed genes in petunia corollas. Each methylated gene averagely carries 1.55 peaks. Among the identified m1A peaks, there are 251 m1A peaks in which the adenines was partly replaced by thymine (T) and/or the reverse transcription stops happened in adenine site, in 199 expressed genes. We found that m1A is enriched in coding sequences with one peaks located immediately after start codons, and a slight negative correlation between methylated genes and gene expression was observed. Totally, ethylene treatment reduced the m1A level of mRNA in petunia corollas. We show that a RNA m1A-methyltransferase, tRNA specific methyltransferase 61A (PhTRMT61A), is an m1A mRNA methyltransferase. PhTRMT61A silencing results in decreased m1A peaks in mRNA in leaves and abnormal leaf development. PhTRMT61A is located to the nucleus. Our results suggest that m1A in mRNA is an important epitranscriptome marker and plays a role in plant development.

2020-08-12 | GSE136921 | GEO

Gene expression analysis during adventitious root formation in petunia cuttings

Project description:In order to identify genes specifically induced during various developmental stages of Adventitious Root (AR) formation (described in Ahkami et al. 2009) in leafy cuttings of Petunia hybrida (line W115) and to describe the series of physiological processes during adventitious rooting, a microarray-based transcriptome analysis in the stem base of the cuttings was conducted. The microarray was described by Breuillin et al. (2010) and included a normalized cDNA library from different time points after taking the cuttings from mother plant. Because physiological processes and molecular changes specifically involved in AR formation were considered as of major interest, rather than those associated with wound responses, a filtration approach was chosen to eliminate primarily wound-responsive genes. This study is mainly focused on changes in transcript abundances of genes related to specific metabolic pathways or cellular events including primary metabolism, membrane transport, cell division or signalling during various phases of AR formation. The overall design of the custom microarray used in this study is described in Breuillin et al. (2010). In brief, a database of 24,816 unigene sequences (including 4,700 ESTs from P. hybrida cutting base and all public available P. hybrida and P. axillaris sequences) was used for construction of a custom microarray.

2014-05-17 | E-GEOD-57752 | biostudies-arrayexpress

Functional characterization of a WRKY transcription factor involved in the regulation of vacuolar transport and flavonoid biosynthesis in petunia

Project description:The WRKY family represents a plant-specific class of zinc-finger transcription factors (TF) well known for regulating abiotic and biotic stress tolerance and also for their emergent role in the control of various developmental and physiological processes (Rushton et al. 2010, Schluttenhofer et al. 2015). The well-characterized AtTTG2, PhPH3 and BnTTG2 are ortholog WRKY proteins with interchangeable functions ascribed to an emergent clade whose regulatory functions are mainly associated with vacuolar metabolism (Li et al. 2015, Verweij et al. 2016, Gonzalez et al. 2016). In particular, PH3 of Petunia hybrida and TTG2 of Arabidopsis thaliana both act in the control of vacuolar pH and trafficking influencing the deposition of secondary metabolites in the petal epidermal cells and in the seed coat, respectively (Verweij et al. 2016, Gonzalez et al. 2016). In this study we functional characterized VvWRKY26 identified as the closest grapevine homolog of PhPH3 and AtTTG2 (Wang et al. 2014; Verweij et al. 2016). VvWRKY26 can fulfil the PH3 function in the regulation of vacuolar pH and impact many aspects of transport when constitutively expressed in petunia ph3 mutant. By a global correlation analysis of gene expression and by transient over-expression in Vitis vinifera cv. Sultana, we showed transcriptomic relationships of VvWRKY26 with many genes related to acidification and trafficking in grapevine. Moreover, our results indicate an involvement in flavonoid pathway mainly in the control in PA biosynthesis in grapevine that is supported by its expression profile. Overall, with the identification of VvWRKY26 our studies might pave the way towards the comprehension of regulatory mechanism underlying the acidification that contributes to the final berry quality traits.

2017-03-08 | GSE86890 | GEO