Integrated analysis of transcriptomic and metabolomic data reveals critical metabolic pathways involved in polyphenol biosynthesis in Nicotiana tabacum under chilling stress
Peilu Zhou
A B E , Qiyao Li A B E , Guangliang Liu A , Na Xu A , Yinju Yang A B , Wenlong Zeng C , Aiguo Chen A D and Shusheng Wang A D
+ Author Affiliations
- Author Affiliations
A Key Laboratory of Tobacco Biology and Processing, Ministry of Agriculture, Tobacco Research Institute of Chinese Academy of Agricultural Sciences, Qingdao 266101, People’s Republic of China.
B Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, People’s Republic of China.
C Longyan Tobacco Agricultural Science Institute, Longyan, Fujian 364000, People’s Republic of China.
D Corresponding authors. Emails: chenaiguo@caas.cn; wangshusheng@caas.cn
E These authors contributed equally to this work.
Functional Plant Biology 46(1) 30-43 https://doi.org/10.1071/FP18099
Submitted: 17 April 2018 Accepted: 15 August 2018 Published: 15 October 2018
Abstract
Chilling stress increases the amount of polyphenols, especially lignin, which protects tobacco (Nicotiana tabacum L. cv. k326) from chilling stress. To clarify the molecular biosynthesis mechanism of the key representative compounds, specifically lignin, RNA sequencing and ultra-high pressure liquid chromatography coupled to quadrupole-time of flight mass spectrometry technologies were used to construct transcriptomic and metabolomic libraries from the leaves of tobacco plants subjected to normal (25°C) and chilling (4°C) temperature treatments. Transcriptomic libraries from the different samples were sequenced, generating more than 40 million raw reads. Among nine samples, metabolomic analysis identified a total of 97 encoding enzymes that function in the key steps of pathways related to polyphenol biosynthesis, where 42 metabolites were also located. An integrated analysis of metabolic and transcriptomic data revealed that most of the intermediate metabolites and enzymes related to lignin biosynthesis were synthesised in the leaves under chilling stress, which suggests that the biosynthesis of lignin plays an important role in the response of tobacco leaves to cold temperatures. In addition, the cold insensitivity of chalcone synthase genes might be considered to be an important rate-limiting factor in the process of precursor substance flow to flavonoid biosynthesis under chilling stress. Furthermore, the upregulated expression of phenylalanine ammonia lyase (PAL), hydroxycinnamoyl transferase (HCT) and cinnamyl-alcohol dehydrogenase (CAD) under chilling stress is the key to an increase in lignin synthesis. This study provides a hypothetical basis for the screening of new active metabolites and the metabolic engineering of polyphenols in tobacco.
Additional keywords: lignin, metabolome, polyphenol metabolic pathway, tobacco, transcriptome.
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