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      De novo transcriptome analysis and glucosinolate profiling in watercress ( Nasturtium officinale R. Br.)

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          Abstract

          Background

          Watercress ( Nasturtium officinale R. Br.) is an aquatic herb species that is a rich source of secondary metabolites such as glucosinolates. Among these glucosinolates, watercress contains high amounts of gluconasturtiin (2-phenethyl glucosinolate) and its hydrolysis product, 2-phennethyl isothiocyanate, which plays a role in suppressing tumor growth. However, the use of N. officinale as a source of herbal medicines is currently limited due to insufficient genomic and physiological information.

          Results

          To acquire precise information on glucosinolate biosynthesis in N. officinale, we performed a comprehensive analysis of the transcriptome and metabolome of different organs of N. officinale. Transcriptome analysis of N. officinale seedlings yielded 69,570,892 raw reads. These reads were assembled into 69,635 transcripts, 64,876 of which were annotated to transcripts in public databases. On the basis of the functional annotation of N. officinale, we identified 33 candidate genes encoding enzymes related to glucosinolate biosynthetic pathways and analyzed the expression of these genes in the leaves, stems, roots, flowers, and seeds of N. officinale. The expression of NoMYB28 and NoMYB29, the main regulators of aliphatic glucosinolate biosynthesis, was highest in the stems, whereas the key regulators of indolic glucosinolate biosynthesis, such as NoDof1.1, NoMYB34, NoMYB51, and NoMYB122, were strongly expressed in the roots. Most glucosinolate biosynthetic genes were highly expressed in the flowers. HPLC analysis enabled us to detect eight glucosinolates in the different organs of N. officinale. Among these glucosinolates, the level of gluconasturtiin was considerably higher than any other glucosinolate in individual organs, and the amount of total glucosinolates was highest in the flower.

          Conclusions

          This study has enhanced our understanding of functional genomics of N. officinale, including the glucosinolate biosynthetic pathways of this plant. Ultimately, our data will be helpful for further research on watercress bio-engineering and better strategies for exploiting its anti-carcinogenic properties.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12864-017-3792-5) contains supplementary material, which is available to authorized users.

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          Most cited references40

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          Streaming fragment assignment for real-time analysis of sequencing experiments

          We present eXpress, a software package for highly efficient probabilistic assignment of ambiguously mapping sequenced fragments. eXpress uses a streaming algorithm with linear run time and constant memory use. It can determine abundances of sequenced molecules in real time, and can be applied to ChIP-seq, metagenomics and other large-scale sequencing data. We demonstrate its use on RNA-seq data, showing greater efficiency than other quantification methods.
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            Deep RNA sequencing at single base-pair resolution reveals high complexity of the rice transcriptome.

            Understanding the dynamics of eukaryotic transcriptome is essential for studying the complexity of transcriptional regulation and its impact on phenotype. However, comprehensive studies of transcriptomes at single base resolution are rare, even for modern organisms, and lacking for rice. Here, we present the first transcriptome atlas for eight organs of cultivated rice. Using high-throughput paired-end RNA-seq, we unambiguously detected transcripts expressing at an extremely low level, as well as a substantial number of novel transcripts, exons, and untranslated regions. An analysis of alternative splicing in the rice transcriptome revealed that alternative cis-splicing occurred in approximately 33% of all rice genes. This is far more than previously reported. In addition, we also identified 234 putative chimeric transcripts that seem to be produced by trans-splicing, indicating that transcript fusion events are more common than expected. In-depth analysis revealed a multitude of fusion transcripts that might be by-products of alternative splicing. Validation and chimeric transcript structural analysis provided evidence that some of these transcripts are likely to be functional in the cell. Taken together, our data provide extensive evidence that transcriptional regulation in rice is vastly more complex than previously believed.
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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
                jeonjin519@gmail.com
                asop_258@naver.com
                jongseok@cnu.ac.kr
                yk-park@lascience.co.kr
                mvalanarasu@gmail.com
                naldhabi@ksu.edu.sa
                +82-42-821-5730 , supark@cnu.ac.kr
                Journal
                BMC Genomics
                BMC Genomics
                BMC Genomics
                BioMed Central (London )
                1471-2164
                23 May 2017
                23 May 2017
                2017
                : 18
                : 401
                Affiliations
                [1 ]ISNI 0000 0001 0722 6377, GRID grid.254230.2, Department of Crop Science, , Chungnam National University, ; 99 Daehak-ro, Yuseong-gu, Daejeon, 34134 Korea
                [2 ]ISNI 0000 0001 0722 6377, GRID grid.254230.2, Department of Horticulture, , Chungnam National University, ; 99 Daehak-ro, Yuseong-gu, Daejeon, 34134 Korea
                [3 ]LAS Inc., 16 Arayuk-ro, Gimpo City, 10136 Korea
                [4 ]ISNI 0000 0004 1773 5396, GRID grid.56302.32, Department of Botany and Microbiology, Addiriyah Chair for Environmental Studies, College of Science, , King Saud University, ; P. O. Box 2455, Riyadh, 11451 Saudi Arabia
                Article
                3792
                10.1186/s12864-017-3792-5
                5442658
                28535746
                796b9dae-6bec-4424-a1e0-87baaa215d0c
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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
                : 25 January 2017
                : 14 May 2017
                Funding
                Funded by: Ministry of Agriculture, Food, and Rural affairs (MAFRA)
                Award ID: IPET(115054-02-1-SB010)
                Award Recipient :
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2017

                Genetics
                nasturtium officinale,watercress,transcriptome,glucosinolates
                Genetics
                nasturtium officinale, watercress, transcriptome, glucosinolates

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