Transcriptome analysis of Polygonum minus reveals candidate genes involved in important secondary metabolic pathways of phenylpropanoids and flavonoids

Background Polygonum minus is an herbal plant in the Polygonaceae family which is rich in ethnomedicinal plants. The chemical composition and characteristic pungent fragrance of Polygonum minus have been extensively studied due to its culinary and medicinal properties. There are only a few transcriptome sequences available for species from this important family of medicinal plants. The limited genetic information from the public expressed sequences tag (EST) library hinders further study on molecular mechanisms underlying secondary metabolite production. Methods In this study, we performed a hybrid assembly of 454 and Illumina sequencing reads from Polygonum minus root and leaf tissues, respectively, to generate a combined transcriptome library as a reference. Results A total of 34.37 million filtered and normalized reads were assembled into 188,735 transcripts with a total length of 136.67 Mbp. We performed a similarity search against all the publicly available genome sequences and found similarity matches for 163,200 (86.5%) of Polygonum minus transcripts, largely from Arabidopsis thaliana (58.9%). Transcript abundance in the leaf and root tissues were estimated and validated through RT-qPCR of seven selected transcripts involved in the biosynthesis of phenylpropanoids and flavonoids. All the transcripts were annotated against KEGG pathways to profile transcripts related to the biosynthesis of secondary metabolites. Discussion This comprehensive transcriptome profile will serve as a useful sequence resource for molecular genetics and evolutionary research on secondary metabolite biosynthesis in Polygonaceae family. Transcriptome assembly of Polygonum minus can be accessed at http://prims.researchfrontier.org/index.php/dataset/transcriptome.

[1]  K. Yaacob Essential Oil of Polygonum minus Huds. , 1990 .

[2]  M. Gómez-Lim,et al.  A Method for Extracting Intact RNA from Fruits Rich in Polysaccharides using Ripe Mango Mesocarp , 1992 .

[3]  X. Huang,et al.  CAP3: A DNA sequence assembly program. , 1999, Genome research.

[4]  B. Seitz,et al.  Matrix polysaccharide precursors in Arabidopsis cell walls are synthesized by alternate pathways with organ-specific expression patterns. , 2000, The Plant journal : for cell and molecular biology.

[5]  K. Shetty Role of proline-linked pentose phosphate pathway in biosynthesis of plant phenolics for functional food and environmental applications: a review , 2004 .

[6]  T. Wetter,et al.  Using the miraEST assembler for reliable and automated mRNA transcript assembly and SNP detection in sequenced ESTs. , 2004, Genome research.

[7]  D. Treutter Significance of flavonoids in plant resistance: a review , 2006 .

[8]  Tania Nolan,et al.  Quantification of mRNA using real-time RT-PCR , 2006, Nature Protocols.

[9]  G. Witzany,et al.  Plant Communication from Biosemiotic Perspective , 2006, Plant signaling & behavior.

[10]  L. Korkina Phenylpropanoids as naturally occurring antioxidants: from plant defense to human health. , 2007, Cellular and molecular biology.

[11]  Y. Lou,et al.  Plant Terpenoids: Biosynthesis and Ecological Functions , 2007 .

[12]  S. Jackson,et al.  Next-generation sequencing technologies and their implications for crop genetics and breeding. , 2009, Trends in biotechnology.

[13]  I. Baldwin,et al.  Jasmonate and ppHsystemin Regulate Key Malonylation Steps in the Biosynthesis of 17-Hydroxygeranyllinalool Diterpene Glycosides, an Abundant and Effective Direct Defense against Herbivores in Nicotiana attenuata[W] , 2010, Plant Cell.

[14]  N. Noor,et al.  Analysis of the Chemical Composition of the Essential Oil of Polygonum minus Huds. Using Two-Dimensional Gas Chromatography-Time-of-Flight Mass Spectrometry (GC-TOF MS) , 2010, Molecules.

[15]  N. Friedman,et al.  Trinity: reconstructing a full-length transcriptome without a genome from RNA-Seq data , 2011, Nature Biotechnology.

[16]  Marcel Martin Cutadapt removes adapter sequences from high-throughput sequencing reads , 2011 .

[17]  C. Cazzonelli Carotenoids in nature: insights from plants and beyond. , 2011, Functional plant biology : FPB.

[18]  Chengying Shi,et al.  Deep sequencing of the Camellia sinensis transcriptome revealed candidate genes for major metabolic pathways of tea-specific compounds , 2011, BMC Genomics.

[19]  Chuan-Yun Li,et al.  KOBAS 2.0: a web server for annotation and identification of enriched pathways and diseases , 2011, Nucleic Acids Res..

[20]  Colin N. Dewey,et al.  RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome , 2011, BMC Bioinformatics.

[21]  Yi Zheng,et al.  iAssembler: a package for de novo assembly of Roche-454/Sanger transcriptome sequences , 2011, BMC Bioinformatics.

[22]  Z. Mohamed-Hussein,et al.  Transcriptome profiling of genes induced by salicylic acid and methyl jasmonate in Polygonum minus , 2012, Molecular Biology Reports.

[23]  L. Ponnala,et al.  Strategies for transcriptome analysis in nonmodel plants. , 2012, American journal of botany.

[24]  N. Wang,et al.  De Novo Transcriptome of Safflower and the Identification of Putative Genes for Oleosin and the Biosynthesis of Flavonoids , 2012, PloS one.

[25]  Z. Mohamed-Hussein,et al.  Flavonoid Biosynthesis Genes Putatively Identified in the Aromatic Plant Polygonum minus via Expressed Sequences Tag (EST) Analysis , 2012, International journal of molecular sciences.

[26]  Jun Mu,et al.  De novo characterization of the root transcriptome of a traditional Chinese medicinal plant Polygonum cuspidatum , 2012, Science China Life Sciences.

[27]  H. Abé,et al.  Identification of natural diterpenes that inhibit bacterial wilt disease in tobacco, tomato and Arabidopsis. , 2012, Plant & cell physiology.

[28]  E. Bornberg-Bauer,et al.  Evaluating Characteristics of De Novo Assembly Software on 454 Transcriptome Data: A Simulation Approach , 2012, PloS one.

[29]  Yimin Wu,et al.  De Novo Transcriptome Assembly in Chili Pepper (Capsicum frutescens) to Identify Genes Involved in the Biosynthesis of Capsaicinoids , 2013, PloS one.

[30]  I. Wilson,et al.  De novo transcriptome sequencing and digital gene expression analysis predict biosynthetic pathway of rhynchophylline and isorhynchophylline from Uncaria rhynchophylla, a non-model plant with potent anti-alzheimer’s properties , 2014, BMC Genomics.

[31]  Liwang Liu,et al.  De novo transcriptome sequencing of radish (Raphanus sativus L.) and analysis of major genes involved in glucosinolate metabolism , 2013, BMC Genomics.

[32]  R. Marsh,et al.  Comparative analysis of de novo transcriptome assembly , 2013, Science China Life Sciences.

[33]  C. Külheim,et al.  Explaining intraspecific diversity in plant secondary metabolites in an ecological context. , 2014, The New phytologist.

[34]  S. Baharum,et al.  Volatile Profiling of Aromatic Traditional Medicinal Plant, Polygonum minus in Different Tissues and Its Biological Activities , 2014, Molecules.

[35]  G. Jensen,et al.  In vitro and ex-vivo cellular antioxidant protection and cognitive enhancing effects of an extract of Polygonum minus Huds (Lineminus™) demonstrated in a Barnes Maze animal model for memory and learning , 2014, BMC Complementary and Alternative Medicine.

[36]  Ortiz-Zuazaga Humberto,et al.  The khmer software package: enabling efficient sequence analysis , 2014 .

[37]  G. Narasimhulu,et al.  THE GENUS POLYGONUM (POLYGONACEAE): AN ETHNOPHARMACOLOGICAL AND PHYTOCHEMICAL PERSPECTIVES -REVIEW , 2014 .

[38]  Björn Usadel,et al.  Trimmomatic: a flexible trimmer for Illumina sequence data , 2014, Bioinform..

[39]  M. M. Mohd Ghazali,et al.  Apoptosis Induction by Polygonum minus Is Related to Antioxidant Capacity, Alterations in Expression of Apoptotic-Related Genes, and S-Phase Cell Cycle Arrest in HepG2 Cell Line , 2014, BioMed research international.

[40]  I. Ismail,et al.  Establishment of Persicaria minor hairy roots and analysis of secreted β-caryophyllene in medium broth , 2014, Plant Cell, Tissue and Organ Culture (PCTOC).

[41]  M. Vikneswaran,et al.  Review on Polygonum minus. Huds, a commonly used food additive in Southeast Asia , 2015, Pharmacognosy research.

[42]  B. Sarangi,et al.  Transcriptome analysis for identification of indigo biosynthesis pathway genes in Polygonum tinctorium , 2015, Biologia.

[43]  Tae-Jin Yang,et al.  Comprehensive analysis of Panax ginseng root transcriptomes , 2015, BMC Plant Biology.

[44]  S. Baharum,et al.  Metabolite profiling reveals temperature effects on the VOCs and flavonoids of different plant populations. , 2016, Plant biology.

[45]  Z. Mohamed-Hussein,et al.  RNA-seq analysis for secondary metabolite pathway gene discovery in Polygonum minus , 2015, Genomics data.