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Enhancement of chlorogenic acid production in hairy roots of Platycodon grandiflorum by over-expression of an Arabidopsis thaliana transcription factor AtPAP1.

ABSTRACT: To improve the production of chlorogenic acid (CGA) in hairy roots of Platycodon grandiflorum, we induced over-expression of Arabidopsis thaliana transcription factor production of anthocyanin pigment (AtPAP1) using an Agrobacterium rhizogenes-mediated transformation system. Twelve hairy root lines showing over-expression of AtPAP1 were generated. In order to investigate the regulation of AtPAP1 on the activities of CGA biosynthetic genes, the expression levels of seven P. grandiflorum CGA biosynthetic genes were analyzed in the hairy root line that had the greatest accumulation of AtPAP1 transcript, OxPAP1-1. The introduction of AtPAP1 increased the mRNA levels of all examined CGA biosynthetic genes and resulted in a 900% up-regulation of CGA accumulation in OxPAP1-1 hairy roots relative to controls. This suggests that P. grandiflorum hairy roots that over-express the AtPAP1 gene are a potential alternative source of roots for the production of CGA.

SUBMITTER: Tuan PA

PROVIDER: S-EPMC4159879 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Enhancement of chlorogenic acid production in hairy roots of Platycodon grandiflorum by over-expression of an Arabidopsis thaliana transcription factor AtPAP1.

Tuan Pham Anh PA Kwon Do Yeon DY Lee Sanghyun S Arasu Mariadhas Valan MV Al-Dhabi Naif Abdullah NA Park Nam Il NI Park Sang Un SU

International journal of molecular sciences 20140822 8

To improve the production of chlorogenic acid (CGA) in hairy roots of Platycodon grandiflorum, we induced over-expression of Arabidopsis thaliana transcription factor production of anthocyanin pigment (AtPAP1) using an Agrobacterium rhizogenes-mediated transformation system. Twelve hairy root lines showing over-expression of AtPAP1 were generated. In order to investigate the regulation of AtPAP1 on the activities of CGA biosynthetic genes, the expression levels of seven P. grandiflorum CGA biosy ...[more]

PMID: 25153629

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Project description:Platycodon grandiflorum is a perennial plant that has been used for medicinal purposes. Specifically, it is widely used in Northern China and Korea for the treatment of various diseases. Terpenoids belong to a group called secondary metabolites and have attracted a wide range of interest. Here, we determined the expressed sequence tag (EST) library of the methyl jasmonate (MeJA)-treated hairy root of P. grandiflorum. In total, 5760 ESTs were obtained, but after deleting the vector sequences and removing poor-quality sequences, a total of 2536 ESTs were attained. Of these, 811 contigs and 1725 singletons were annotated. The data were further analyzed with a focus on the gene families of the terpenoid biosynthetic pathway (TBP). We identified and characterized four TBP genes; among these were three full-length cDNAs encoding PgHMGS, PgMK, and PgMVD, whereas PgHMGR had a partial sequence based on the EST library database. Phylogenetic analysis and a pairwise identity matrix showed that these identified genes were closely related to those of other higher plants. Moreover, the tertiary structure and multiple alignment analysis showed that several distinct conserved motifs were present. Quantitative reverse transcription-polymerase chain reaction results revealed that TBP genes were constitutively expressed in all organs of P. grandiflorum, while the expression of transcript levels of these genes varied distinctly among the organs. Additionally, the total amount of platycosides was highly detected in the root, accumulating in concentrations 7.8 and 2.6 times higher than in the hairy root and stem, respectively, and 1.4 times higher than in the leaf and flower. The highest amount of total phytosterols was found to accumulate in the leaves at 9.3, 9.1, 1.8, and 1.6 times higher than that of the stem, root, hairy root, and flower, respectively. After the hairy root was exposed to the MeJA treatment, the transcript levels of PgHMGS, PgHMGR, PgMK, and PgMVD had significantly increased. The highest level of transcript accumulation occurred at 3 h after initial exposure for most of the genes. Meanwhile, triterpene saponin accumulation increased with the increase in the time of exposure, and at 48 h after initial exposure, the total saponin content was the highest recorded.

Project description:BackgroundCampanulaceae species are known to have highly rearranged plastid genomes lacking the acetyl-CoA carboxylase (ACC) subunit D gene (accD), and instead have a nuclear (nr)-accD. Plastid genome information has been thought to depend on studies concerning Trachelium caeruleum and genome announcements for Adenophora remotiflora, Campanula takesimana, and Hanabusaya asiatica. RNA editing information for plastid genes is currently unavailable for Campanulaceae. To understand plastid genome evolution in Campanulaceae, we have sequenced and characterized the chloroplast (cp) genome and nr-accD of Platycodon grandiflorum, a basal member of Campanulaceae.ResultsWe sequenced the 171,818 bp cp genome containing a 79,061 bp large single-copy (LSC) region, a 42,433 bp inverted repeat (IR) and a 7840 bp small single-copy (SSC) region, which represents the cp genome with the largest IR among species of Campanulaceae. The genome contains 110 genes and 18 introns, comprising 77 protein-coding genes, four RNA genes, 29 tRNA genes, 17 group II introns, and one group I intron. RNA editing of genes was detected in 18 sites of 14 protein-coding genes. Platycodon has an IR containing a 3' rps12 operon, which occurs in the middle of the LSC region in four other species of Campanulaceae (T. caeruleum, A. remotiflora, C. takesimana, and H. asiatica), but lacks accD, clpP, infA, and rpl23, as has been found in these four species. Platycodon nr-accD contains about 3.2 kb intron between nr-accD.e1 and nr-accD.e2 at the same insertion point as in other Campanulaceae. The phylogenies of the plastid genomes and accD show that Platycodon is basal in the Campanulaceae clade, indicating that IR disruption in Campanulaceae occurred after the loss of accD, clpP, infA, and rpl23 in the cp genome, which occurred during plastid evolution in Campanulaceae.ConclusionsThe plastid genome of P. grandiflorum lacks the rearrangement of the IR found in T. caeruleum, A. remotiflora, C. takesimana, and H. asiatica. The absence of accD, clpP, infA, and rpl23 in the plastid genome is a synapomorphic characteristic of Campanulaceae. The chloroplast genome phylogeny supports the hypothesis that chloroplast genomic arrangement occurred after accD nuclear transfer and loss of the four genes in the plastid of early Campanulaceae as a lineage of asterids.

Dataset Information

Enhancement of chlorogenic acid production in hairy roots of Platycodon grandiflorum by over-expression of an Arabidopsis thaliana transcription factor AtPAP1.

Publications

Enhancement of chlorogenic acid production in hairy roots of Platycodon grandiflorum by over-expression of an Arabidopsis thaliana transcription factor AtPAP1.

Similar Datasets

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