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      De novo transcriptome analysis of petal senescence in Gardenia jasminoides Ellis

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          Abstract

          Background

          The petal senescence of ethylene insensitive species has not been investigated thoroughly while little is known about the temporal and tissue specific expression patterns of transcription factors (TFs) in this developmental process. Even less is known on flower senescence of the ornamental pot plant Gardenia jasminoides, a non climacteric flower with significant commercial value.

          Results

          We initiated a de novo transcriptome study to investigate the petal senescence in four developmental stages of cut gardenia flowers considering that the visible symptoms of senescence appear within 4 days of flower opening. De novo assembly of transcriptome sequencing resulted in 102,263 contigs with mean length of 360 nucleotides that generated 57,503 unigenes. These were further clustered into 20,970 clusters and 36,533 singletons. The comparison of the consecutive developmental stages resulted in 180 common, differentially expressed unigenes. A large number of Simple Sequence Repeats were also identified comprising a large number of dinucleotides and trinucleotides. The prevailing families of differentially expressed TFs comprise the AP2/EREBP, WRKY and the bHLH. There are 81 differentially expressed TFs when the symptoms of flower senescence become visible with the most prevailing being the WRKY family with 19 unigenes. No other WRKY TFs had been identified up to now in petal senescence of ethylene insensitive species. A large number of differentially expressed genes were identified at the initiation of visible symptoms of senescence compared to the open flower stage indicating a significant shift in the expression profiles which might be coordinated by up-regulated and/or down-regulated TFs. The expression of 16 genes that belong to the TF families of WRKY, bHLH and the ethylene sensing pathway was validated using qRT – PCR.

          Conclusion

          This de novo transcriptome analysis resulted in the identification of TFs with specific temporal expression patterns such as two WRKYs and one bHLH, which might play the role of senescence progression regulators. Further research is required to investigate their role in gardenia flowers in order to develop tools to delay petal senescence.

          Electronic supplementary material

          The online version of this article (doi:10.1186/1471-2164-15-554) contains supplementary material, which is available to authorized users.

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          Most cited references64

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          Functional analysis of an Arabidopsis transcription factor, DREB2A, involved in drought-responsive gene expression.

          Transcription factors DREB1A/CBF3 and DREB2A specifically interact with cis-acting dehydration-responsive element/C-repeat (DRE/CRT) involved in cold and drought stress-responsive gene expression in Arabidopsis thaliana. Intact DREB2A expression does not activate downstream genes under normal growth conditions, suggesting that DREB2A requires posttranslational modification for activation, but the activation mechanism has not been clarified. DREB2A domain analysis using Arabidopsis protoplasts identified a transcriptional activation domain between residues 254 and 335, and deletion of a region between residues 136 and 165 transforms DREB2A to a constitutive active form. Overexpression of constitutive active DREB2A resulted in significant drought stress tolerance but only slight freezing tolerance in transgenic Arabidopsis plants. Microarray and RNA gel blot analyses revealed that DREB2A regulates expression of many water stress-inducible genes. However, some genes downstream of DREB2A are not downstream of DREB1A, which also recognizes DRE/CRT but functions in cold stress-responsive gene expression. Synthetic green fluorescent protein gave a strong signal in the nucleus under unstressed control conditions when fused to constitutive active DREB2A but only a weak signal when fused to full-length DREB2A. The region between DREB2A residues 136 and 165 plays a role in the stability of this protein in the nucleus, which is important for protein activation.
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            Plant responses to ethylene gas are mediated by SCF(EBF1/EBF2)-dependent proteolysis of EIN3 transcription factor.

            Plants use ethylene gas as a signal to regulate myriad developmental processes and stress responses. The Arabidopsis EIN3 protein is a key transcription factor mediating ethylene-regulated gene expression and morphological responses. Here, we report that EIN3 protein levels rapidly increase in response to ethylene and this response requires several ethylene-signaling pathway components including the ethylene receptors (ETR1 and EIN4), CTR1, EIN2, EIN5, and EIN6. In the absence of ethylene, EIN3 is quickly degraded through a ubiquitin/proteasome pathway mediated by two F box proteins, EBF1 and EBF2. Plants containing mutations in either gene show enhanced ethylene response by stabilizing EIN3, whereas efb1 efb2 double mutants show constitutive ethylene phenotypes. Plants overexpressing either F box gene display ethylene insensitivity and destabilization of EIN3 protein. These results reveal that a ubiquitin/proteasome pathway negatively regulates ethylene responses by targeting EIN3 for degradation, and pinpoint EIN3 regulation as the key step in the response to ethylene.
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              De Novo Assembly of Chickpea Transcriptome Using Short Reads for Gene Discovery and Marker Identification

              Chickpea ranks third among the food legume crops production in the world. However, the genomic resources available for chickpea are still very limited. In the present study, the transcriptome of chickpea was sequenced with short reads on Illumina Genome Analyzer platform. We have assessed the effect of sequence quality, various assembly parameters and assembly programs on the final assembly output. We assembled ∼107million high-quality trimmed reads using Velvet followed by Oases with optimal parameters into a non-redundant set of 53 409 transcripts (≥100 bp), representing about 28 Mb of unique transcriptome sequence. The average length of transcripts was 523 bp and N50 length of 900 bp with coverage of 25.7 rpkm (reads per kilobase per million). At the protein level, a total of 45 636 (85.5%) chickpea transcripts showed significant similarity with unigenes/predicted proteins from other legumes or sequenced plant genomes. Functional categorization revealed the conservation of genes involved in various biological processes in chickpea. In addition, we identified simple sequence repeat motifs in transcripts. The chickpea transcripts set generated here provides a resource for gene discovery and development of functional molecular markers. In addition, the strategy for de novo assembly of transcriptome data presented here will be helpful in other similar transcriptome studies.
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                Author and article information

                Contributors
                gtsanak@agro.auth.gr
                manioudaki@maich.gr
                econe@agro.auth.gr
                panagiot@maich.gr
                Journal
                BMC Genomics
                BMC Genomics
                BMC Genomics
                BioMed Central (London )
                1471-2164
                4 July 2014
                4 July 2014
                2014
                : 15
                : 1
                : 554
                Affiliations
                [ ]Department of Horticultural Genetics & Biotechnology, Mediterranean Agronomic Institute of Chania (MAICh), Crete, Greece
                [ ]School of Agriculture, Aristotle University of Thessaloniki, Thessaloniki, Greece
                [ ]Mediterranean Agronomic Institute of Chania (MAICh), Alsyllio Agrokepio, 1 Makedonias str, PO Box 85 Chania, 73100 Crete Greece
                Article
                6265
                10.1186/1471-2164-15-554
                4108791
                24993183
                b641e44f-9d8a-4b17-9534-689c0121b764
                © Tsanakas et al.; licensee BioMed Central Ltd. 2014

                This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 19 February 2014
                : 11 June 2014
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2014

                Genetics
                gardenia,transcriptome,ssr markers,transcription factors,petal senescence
                Genetics
                gardenia, transcriptome, ssr markers, transcription factors, petal senescence

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