Dynamic analysis of gene expression and determination of chemicals in agarwood in Aquilaria sinensis

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ORIGINAL PAPER

Dynamic analysis of gene expression and determination of chemicals in agarwood in Aquilaria sinensis Zeqing Wu1 • Wanzhen Liu2 • Jing Li3 • Liangwen Yu2 • Li Lin3

Received: 9 April 2018 / Accepted: 5 July 2018  The Author(s) 2019, corrected publication 2019

Abstract Agarwood is the resinous heartwood of Aquilaria species. However, low yields and high costs of existing stimulation methods have led to the need for new techniques to produce agarwood rapidly and effectively. We developed a biological agarwood-inducing technique (Agar-Bit) that produces high yields and quality within 6 months. To better understand agarwood formation by Agar-Bit, dynamic gene expressions of key synthetases pathways of sesquiterpenes and chalcone- related enzymes at different times were determined after both Agar-Bit and the traditional burning chisel drilling (BCD) stimulation on Aquilaria sinensis trees. The qRT-PCR results show that some characteristic synthase genes were expressed at greatly different levels and times compared with the

Project funding: The work was supported by Grant Number NDRC2011-51 from the National Development and Reform Commission (NDRC), the Office of New High-Tech Industrial Development. This work was also supported by Grant Number 18A36002 from Key project of institutions of colleges and universities in Henan province, Henan Education Department. The online version is available at http://www.springerlink.com. Corresponding editor: Tao Xu. & Zeqing Wu [email protected] 1

College of Pharmacy, Xinxiang Medical University, Xinxiang 453003, Henan, People’s Republic of China

2

College of Sports and Health, Guangzhou University of Chinese Medicine, Guangzhou 510006, Guangdong, People’s Republic of China

3

College of Chinese Materia Medical, Guangzhou University of Chinese Medicine, Guangzhou 510006, Guangdong, People’s Republic of China

controls. For the Agar-Bit technology, main changes were after the 3rd or 5th month, while BCD expression clearly changed at the 5th month. Essential oils and total chromone contents were simultaneously determined. In the Agar-Bit group, both were higher and similar to natural levels. The Agar-Bit methodology is a new option for producing agarwood as demonstrated by genetic and chemical aspects. The differences in gene expression within 6 months for both groups indicates that the mechanisms of the two methods are different. These findings provide information on genetic variation during the process of agarwood formation. Keywords Agarwood  Biological agarwood-inducing technique (Agar-Bit)  Essential oils  Gene expression  Total chromone

Introduction Agarwood, resinous trunks or branches from Aquilaria trees, is widely used in perfume manufacturing, the production of incense and in traditional Chinese medicine. However, agarwood is only harvested from wounded trees. In nature, agarwood is formed by insect feeding, lightning strikes or storm damage and result in low production, keeping prices high. There was previously a problem for natural Aquilaria becaus