Deep sequencing reveals transcriptome re-programming of Taxus × media cells to the elicitation with methyl jasmonate

PLoS One. 2013 Apr 30;8(4):e62865. doi: 10.1371/journal.pone.0062865. Print 2013.

Abstract

Background: Plant cell culture represents an alternative source for producing high-value secondary metabolites including paclitaxel (Taxol®), which is mainly produced in Taxus and has been widely used in cancer chemotherapy. The phytohormone methyl jasmonate (MeJA) can significantly increase the production of paclitaxel, which is induced in plants as a secondary metabolite possibly in defense against herbivores and pathogens. In cell culture, MeJA also elicits the accumulation of paclitaxel; however, the mechanism is still largely unknown.

Methodology/principal findings: To obtain insight into the global regulation mechanism of MeJA in the steady state of paclitaxel production (7 days after MeJA addition), especially on paclitaxel biosynthesis, we sequenced the transcriptomes of MeJA-treated and untreated Taxus × media cells and obtained ∼ 32.5 M high quality reads, from which 40,348 unique sequences were obtained by de novo assembly. Expression level analysis indicated that a large number of genes were associated with transcriptional regulation, DNA and histone modification, and MeJA signaling network. All the 29 known genes involved in the biosynthesis of terpenoid backbone and paclitaxel were found with 18 genes showing increased transcript abundance following elicitation of MeJA. The significantly up-regulated changes of 9 genes in paclitaxel biosynthesis were validated by qRT-PCR assays. According to the expression changes and the previously proposed enzyme functions, multiple candidates for the unknown steps in paclitaxel biosynthesis were identified. We also found some genes putatively involved in the transport and degradation of paclitaxel. Potential target prediction of miRNAs indicated that miRNAs may play an important role in the gene expression regulation following the elicitation of MeJA.

Conclusions/significance: Our results shed new light on the global regulation mechanism by which MeJA regulates the physiology of Taxus cells and is helpful to understand how MeJA elicits other plant species besides Taxus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetates / pharmacology*
  • Cell Line
  • Cells, Cultured
  • Computational Biology
  • Cyclopentanes / metabolism
  • Cyclopentanes / pharmacology*
  • Databases, Genetic
  • Ethylenes / biosynthesis
  • Gene Expression Regulation, Plant / drug effects
  • Genes, Plant
  • High-Throughput Nucleotide Sequencing*
  • MicroRNAs / genetics
  • MicroRNAs / metabolism
  • Molecular Sequence Annotation
  • Oxylipins / metabolism
  • Oxylipins / pharmacology*
  • Paclitaxel / biosynthesis
  • Plant Cells / drug effects*
  • Plant Cells / metabolism*
  • Plant Growth Regulators / pharmacology*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Reproducibility of Results
  • Taxus / cytology*
  • Terpenes / metabolism
  • Transcriptome*

Substances

  • Acetates
  • Cyclopentanes
  • Ethylenes
  • MicroRNAs
  • Oxylipins
  • Plant Growth Regulators
  • RNA, Messenger
  • Terpenes
  • jasmonic acid
  • methyl jasmonate
  • ethylene
  • Paclitaxel

Grants and funding

This work is supported by a grant from the National Natural Science Foundation of China (31170628) and a startup grant from Kunming Institute of Botany, Chinese Academy of Sciences, to the Group of Molecular Ecology, a grant from Yunnan Recruitment Program of Experts in Sciences (2012HA016), and a grant from Thousand Youth Talents Program. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.