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Protoplasma

, Volume 252, Issue 3, pp 813–824 | Cite as

Gene transcript profiles of the TIA biosynthetic pathway in response to ethylene and copper reveal their interactive role in modulating TIA biosynthesis in Catharanthus roseus

  • Ya-Jie Pan
  • Jia Liu
  • Xiao-Rui Guo
  • Yuan-Gang Zu
  • Zhong-Hua Tang
Original Article

Abstract

Research on transcriptional regulation of terpenoid indole alkaloid (TIA) biosynthesis of the medicinal plant, Catharanthus roseus, has largely been focused on gene function and not clustering analysis of multiple genes at the transcript level. Here, more than ten key genes encoding key enzyme of alkaloid synthesis in TIA biosynthetic pathways were chosen to investigate the integrative responses to exogenous elicitor ethylene and copper (Cu) at both transcriptional and metabolic levels. The ethylene-induced gene transcripts in leaves and roots, respectively, were subjected to principal component analysis (PCA) and the results showed the overall expression of TIA pathway genes indicated as the Q value followed a standard normal distribution after ethylene treatments. Peak gene expression was at 15–30 μM of ethephon, and the pre-mature leaf had a higher Q value than the immature or mature leaf and root. Treatment with elicitor Cu found that Cu up-regulated overall TIA gene expression more in roots than in leaves. The combined effects of Cu and ethephon on TIA gene expression were stronger than their separate effects. It has been documented that TIA gene expression is tightly regulated by the transcriptional factor (TF) ethylene responsive factor (ERF) and mitogen-activated protein kinase (MAPK) cascade. The loading plot combination with correlation analysis for the genes of C. roseus showed that expression of the MPK gene correlated with strictosidine synthase (STR) and strictosidine b-d-glucosidase(SGD). In addition, ERF expression correlated with expression of secologanin synthase (SLS) and tryptophan decarboxylase (TDC), specifically in roots, whereas MPK and myelocytomatosis oncogene (MYC) correlated with STR and SGD genes. In conclusion, the ERF regulates the upstream pathway genes in response to heavy metal Cu mainly in C. roseus roots, while the MPK mainly participates in regulating the STR gene in response to ethylene in pre-mature leaf. Interestingly, the change in TIA accumulation does not correlate with expression of the associated genes. Our previous research found significant accumulation of vinblastine in response to high concentration of ethylene and Cu suggesting the involvement of posttranscriptional and posttranslational mechanisms in a spatial and temporal manner. In this study, meta-analysis reveals ERF and MPK form a positive feedback loop connecting two pathways actively involved in response of TIA pathway genes to ethylene and copper in C. roseus.

Keywords

Catharanthus roseus Copper Ethylene Terpenoid indole alkaloids Transcript clustering 

Abbreviations

AS

Anthranilate synthase

DAT

Acetyl-CoA: 4-O-deacetylvindoline4-O-acetyl-transferase

D4H

Desacetoxyvindoline-4-hydroxylase

ERF

Ethylene responsive factor

G10H

Geraniol 10-hydroxylase

MAPK

Mitogen-activated protein kinase

MeJA

Methyl-jasmonate

MYC

Myelocytomatosis oncogen

ORCA

Octadecanoid-responsive Catharanthus AP2-domain

PCA

Principal component analysis

qRT-PCR

Quantitative real-time reverse transcription

RPS

Ribosomal protein subunit

SGD

Strictosidine b-d-glucosidase

SLS

Secologanin synthase

STR

Strictosidine synthase

TDC

Tryptophan decarboxylase

TF

Transcriptional factor

TIA

Terpenoid indole alkaloid

T16H

Tabersonine-16-hydroxylase

Notes

Acknowledgments

We sincerely thank Matthew Paul from Rothamsted Research and Ling Yuan from University of Kentucky for their constructive and critical comments on this manuscript. This study was financially supported by the Fundamental Research Funds for the Central Universities (2572014DA01) and National Natural Science Foundation of China (31400337 and 31370007).

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Copyright information

© Springer-Verlag Wien 2014

Authors and Affiliations

  • Ya-Jie Pan
    • 1
  • Jia Liu
    • 1
  • Xiao-Rui Guo
    • 1
  • Yuan-Gang Zu
    • 1
  • Zhong-Hua Tang
    • 1
  1. 1.The Key Laboratory of Plant EcologyNortheast Forestry UniversityHarbinChina

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