Triterpenoid content and expression of triterpenoid biosynthetic genes in birch ( Betula platyphylla Suk) treated with 5
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ORIGINAL PAPER
Triterpenoid content and expression of triterpenoid biosynthetic genes in birch (Betula platyphylla Suk) treated with 5-azacytidine Fansuo Zeng1,2 • Xiaoyi Li2 • Rui Qie3 • Leilei Li2 • Minghao Ma2 Yaguang Zhan1,2
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Received: 3 June 2018 / Accepted: 3 October 2018 Northeast Forestry University 2019
Abstract DNA methylation is widespread in plants and associated with plant development and defense mechanisms. However, the relationship between DNA methylation and plant secondary metabolism has rarely been reported. Here, when birch suspension cells were treated with 5-azacytidine (5-azaC), which blocks DNA methylation, triterpenoid accumulation was significantly promoted and antioxidant and defense enzymatic activity changed. For studying triterpenoid accumulation, 0.1 mM azaC was optimal. A qRT-PCR assay revealed increased expression of genes encoding key triterpenoid biosynthetic enzymes. Evaluation of methylation polymorphisms at CCGG sites showed that the methylation level was lower in cells treated with 5-azaC. These results demonstrated that 5-azaC treatment led to an increase in the production of triterpenoids in cell cultures through a mechanism that involved in DNA methylation, which resulted in the induction of genes encoding the key enzymes. The study provides Project funding: The work was supported by The Fundamental Research Funds for the Central Universities (2572017EA05) and the National Natural Science Foundation of China (31870588). The online version is available at http://www.springerlink.com. Corresponding editor: Tao Xu. & Yaguang Zhan [email protected] 1
State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040, People’s Republic of China
2
College of Life Science, Northeast Forestry University, Harbin 150040, People’s Republic of China
3
First Affiliated Hospital, Heilongjiang University of Chinese Medicine, Harbin 150040, People’s Republic of China
evidence of a relationship between DNA methylation and regulation of secondary metabolism. Keywords 5-Azacytidine Methylation Suspension cells Triterpenoids biosynthesis
Introduction Birch (Betula platyphylla Suk) is indisputably one of the most important commercial tree species native to China for its high quality industrial timber and for plywood, paper, baking, furniture, pulp material among other uses (Li et al. 1995). The bark also has important medicinal value for treating yellow gangrene, breast carbuncle, scabies and other diseases (Krasutsky 2006; Guo et al. 2017). In recent years, the pharmacological effect of birch bark, which has a complex composition, has received global attention. Some of the main active components in the bark are triterpenoids (Eom et al. 2016) such as oleanolic acid, ursolic acid and betulinic acid, which have outstanding antiviral and antitumor properties (Dzubak et al. 2006; Petronelli et al. 2009; Yadav et al. 2014; Lomkova et al. 2016). Currently, triterpenes are mainly extracted and separated from the bark of B. platyphylla, but because bi
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