biologia plantarum

Biologia plantarum 59:695-700, 2015 | DOI: 10.1007/s10535-015-0541-y

Identification of hydrogen peroxide responsive ESTs involved in phenylethanoid glycoside biosynthesis in Cistanche salsa cell culture

J. Chen¹, Y. -X. Yan¹, Z. -G. Guo^1,*

¹ Institute of Biochemical Engineering, Department of Chemical Engineering, Tsinghua University, Beijing, P.R.China

Hydrogen peroxide is an effective abiotic elicitor that can induce secondary metabolite biosynthesis in plants. We show that in cell suspension culture of a salt-tolerant medicinal plant Cistanche salsa, the production of bioactive components phenylethanoid glycosides (PeGs) was increased after an H₂O₂ treatment. To identify genes related to PeGs biosynthesis affected by H₂O₂, we constructed a suppression subtractive hybridization library of H₂O₂ responsive genes using a C. salsa cell line and identified 105 expressed sequence tags (ESTs) and 85 genes. EST library functional annotation and gene ontology analyses showed genes related to various stress responses, biosynthesis of secondary metabolites, and transcriptional regulation. Among them we identified two genes related to the PeGs biosynthesis pathway (4-coumarate coenzyme A ligase and cinnamate 4-hydroxylase), and two WRKY type transcription factors. The expressions of selected genes after the H₂O₂ treatment were analyzed by RT-qPCR. An early increased transcription of PeG biosynthesis pathway genes after the treatment revealed that H₂O₂ induced PeGs biosynthesis via up-regulation of its key genes.

Keywords: cinnamate-4-hydroxylase; 4-coumarate coenzyme A ligase; suppression subtractive hybridization

Subjects: hydrogen peroxide; expressed sequence tags; suppression subtractive hybridization; cinnamate 4-hydroxylase; 4-coumarate: coenzyme A ligase; phenylethanoid glycosides

Received: August 3, 2014; Revised: January 27, 2015; Accepted: February 10, 2015; Published: December 1, 2015  Show citation