Transcriptomic landscape of Pueraria lobata demonstrates potential for phytochemical study

Pueraria lobata (Willd.) Ohwi has a long and broad application in the treatment of disease. However, in the US and EU, it is treated as a notorious weed. The information to be gained from decoding the deep transcriptome profile would facilitate further research on P. lobata. In this study, more than 93 million fastq format reads were generated by Illumina’s next-generation sequencing approach using five types of P. lobata tissue, followed by CLC de novo assembly methods, ultimately yielding about 83,041 contigs in total. Then BLASTx similarity searches against the NCBI NR database and UniProtKB database were conducted. Once the duplicates among BLASTx hits were eliminated, ID mapping against the UniProt database was conducted online to retrieve Gene Ontology information. In search of the putative genes relevant to essential biosynthesis pathways, all 1,348 unique enzyme commission numbers were used to map pathways against the Kyoto Encyclopedia of Genes and Genomes. Enzymes related to the isoflavonoid and flavonoid biosynthesis pathways were focused for detailed investigation and subsequently, quantitative real-time reverse transcription polymerase chain reaction was conducted for biological validation. Metabolites of interest, puerarin and daidzin were studied by HPLC. The findings in this report may serve as a footstone for further research into this promising medicinal plant.

[1]  Hideyuki Suzuki,et al.  Transcriptome Analysis of Nine Tissues to Discover Genes Involved in the Biosynthesis of Active Ingredients in Sophora flavescens. , 2015, Biological & pharmaceutical bulletin.

[2]  S. Powles,et al.  RNA-Seq transcriptome analysis to identify genes involved in metabolism-based diclofop resistance in Lolium rigidum. , 2014, The Plant journal : for cell and molecular biology.

[3]  Kazuki Saito,et al.  The flavonoid biosynthetic pathway in Arabidopsis: structural and genetic diversity. , 2013, Plant physiology and biochemistry : PPB.

[4]  E. Grotewold,et al.  Identification of a Bifunctional Maize C- and O-Glucosyltransferase* , 2013, The Journal of Biological Chemistry.

[5]  Kazuki Saito,et al.  Phytochemical genomics--a new trend. , 2013, Current opinion in plant biology.

[6]  Jordan A. Ramilowski,et al.  Glycyrrhiza uralensis transcriptome landscape and study of phytochemicals. , 2013, Plant & cell physiology.

[7]  Kazuki Saito,et al.  Phytochemical genomics on the way. , 2013, Plant & cell physiology.

[8]  Zhengwei Zhu,et al.  CD-HIT: accelerated for clustering the next-generation sequencing data , 2012, Bioinform..

[9]  P. Casati,et al.  Flavonoids: biosynthesis, biological functions, and biotechnological applications , 2012, Front. Plant Sci..

[10]  S. Glémin,et al.  Patterns and Evolution of Nucleotide Landscapes in Seed Plants[W] , 2012, Plant Cell.

[11]  A. Conesa,et al.  Differential expression in RNA-seq: a matter of depth. , 2011, Genome research.

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

[13]  Kelvin Chan,et al.  Kudzu root: traditional uses and potential medicinal benefits in diabetes and cardiovascular diseases. , 2011, Journal of ethnopharmacology.

[14]  Shigehiko Kanaya,et al.  Dynamics of time-lagged gene-to-metabolite networks of Escherichia coli elucidated by integrative omics approach. , 2011, Omics : a journal of integrative biology.

[15]  Hanbo Chen,et al.  VennDiagram: a package for the generation of highly-customizable Venn and Euler diagrams in R , 2011, BMC Bioinformatics.

[16]  S. Follak Potential distribution and environmental threat of Pueraria lobata , 2011, Central European Journal of Biology.

[17]  R. Dixon,et al.  A genomic approach to isoflavone biosynthesis in kudzu (Pueraria lobata) , 2011, Planta.

[18]  S. Bahn,et al.  Identification and testing of superior reference genes for a starting pool of transcript normalization in Arabidopsis. , 2010, Plant & cell physiology.

[19]  E. Miadoková Isoflavonoids – an overview of their biological activities and potential health benefits , 2009, Interdisciplinary toxicology.

[20]  Siu-Ming Yiu,et al.  SOAP2: an improved ultrafast tool for short read alignment , 2009, Bioinform..

[21]  Steven J. M. Jones,et al.  Abyss: a Parallel Assembler for Short Read Sequence Data Material Supplemental Open Access , 2022 .

[22]  H. Puschmann,et al.  The C-Glycosylation of Flavonoids in Cereals*♦ , 2009, The Journal of Biological Chemistry.

[23]  Jianqiu Zheng,et al.  Protective effect of puerarin on diabetic retinopathy in rats , 2009, Molecular Biology Reports.

[24]  E. Liu,et al.  Next-generation DNA sequencing of paired-end tags (PET) for transcriptome and genome analyses. , 2009, Genome research.

[25]  R. Dixon,et al.  Regioselective synthesis of plant (iso)flavone glycosides in Escherichia coli , 2008, Applied Microbiology and Biotechnology.

[26]  Thomas D. Schmittgen,et al.  Analyzing real-time PCR data by the comparative CT method , 2008, Nature Protocols.

[27]  W. Cherdshewasart,et al.  Correlation of antioxidant activity and major isoflavonoid contents of the phytoestrogen-rich Pueraria mirifica and Pueraria lobata tubers. , 2008, Phytomedicine : international journal of phytotherapy and phytopharmacology.

[28]  W. Cherdshewasart,et al.  Major isoflavonoid contents of the phytoestrogen rich-herb Pueraria mirifica in comparison with Pueraria lobata. , 2007, Journal of pharmaceutical and biomedical analysis.

[29]  Lin Fang,et al.  WEGO: a web tool for plotting GO annotations , 2006, Nucleic Acids Res..

[30]  J. Bailey-Serres,et al.  mRNA sequence features that contribute to translational regulation in Arabidopsis , 2005, Nucleic acids research.

[31]  Lijuan Cong,et al.  Characterization of soybean genomic features by analysis of its expressed sequence tags , 2004, Theoretical and Applied Genetics.

[32]  J. Zhang,et al.  Determination of puerarin, daidzein and rutin in Pueraria lobata (Wild.) Ohwi by capillary electrophoresis with electrochemical detection. , 2001, Journal of chromatography. A.

[33]  G. Gessa,et al.  Potential use of medicinal plants in the treatment of alcoholism. , 2000, Fitoterapia.

[34]  M. Ashburner,et al.  Gene Ontology: tool for the unification of biology , 2000, Nature Genetics.

[35]  O. Yu,et al.  Identification and expression of isoflavone synthase, the key enzyme for biosynthesis of isoflavones in legumes , 2000, Nature Biotechnology.

[36]  R. Dixon,et al.  Molecular characterization of the enzyme catalyzing the aryl migration reaction of isoflavonoid biosynthesis in soybean. , 1999, Archives of biochemistry and biophysics.

[37]  A. Guckert,et al.  Production of daidzein by callus cultures of Psoralea species and comparison with plants , 1998, Plant Cell, Tissue and Organ Culture.

[38]  B. Vallee,et al.  Kudzu root: an ancient Chinese source of modern antidipsotropic agents. , 1998, Phytochemistry.

[39]  T. L. Graham,et al.  Flavonoid and isoflavonoid distribution in developing soybean seedling tissues and in seed and root exudates. , 1991, Plant physiology.

[40]  G. Franz,et al.  Biosynthesis of Vitexin and Isovitexin: Enzymatic Synthesis of the C-Glucosylflavones Vitexin and Isovitexin with an Enzyme Preparation from Fagopyrum esculentum M. Seedlings , 1987 .

[41]  C. Lindgren,et al.  The Biology of Invasive Alien Plants in Canada. 12. Pueraria montana var. lobata (Willd.) Sanjappa & Predeep , 2013, Canadian Journal of Plant Science.

[42]  M. Gerstein,et al.  RNA-Seq: a revolutionary tool for transcriptomics , 2009, Nature Reviews Genetics.

[43]  H. Kosaka,et al.  Antimutagenic activity of isoflavone from Pueraria lobata. , 2001, Journal of agricultural and food chemistry.

[44]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .

[45]  G. Franz,et al.  Biosynthesis of Vitexin and Isovitexin , 1987 .