title: Transgenic analysis reveals LeACS-1 as a positive regulator of ethylene-induced shikonin biosynthesis in Lithospermum erythrorhizon hairy roots.
abstract: The phytohormone ethylene (ET) is a crucial signaling molecule that induces the biosynthesis of shikonin and its derivatives in Lithospermum erythrorhizon shoot cultures. However, the molecular mechanism and the positive regulators involved in this physiological process are largely unknown. In this study, the function of LeACS-1, a key gene encoding the 1-aminocyclopropane-1-carboxylic acid synthase for ET biosynthesis in L. erythrorhizon hairy roots, was characterized by using overexpression and RNA interference (RNAi) strategies. The results showed that overexpression of LeACS-1 significantly increased endogenous ET concentration and shikonin production, consistent with the up-regulated genes involved in ET biosynthesis and transduction, as well as the genes related to shikonin biosynthesis. Conversely, RNAi of LeACS-1 effectively decreased endogenous ET concentration and shikonin production and down-regulated the expression level of above genes. Correlation analysis showed a significant positive linear relationship between ET concentration and shikonin production. All these results suggest that LeACS-1 acts as a positive regulator of ethylene-induced shikonin biosynthesis in L. erythrorhizon hairy roots. Our work not only gives new insights into the understanding of the relationship between ET and shikonin biosynthesis, but also provides an efficient genetic engineering target gene for secondary metabolite production in non-model plant L. erythrorhizon.
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, NJU-NJFU Joint Institute of Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing, 210093, People's Republic of China.|Jiangsu University of Science and Technology, Zhenjiang, 212003, People's Republic of China.|Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China.
authors.forename: Rongjun
authors.initials: R
authors.lastname: Fang
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, NJU-NJFU Joint Institute of Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing, 210093, People's Republic of China.|Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China.
authors.forename: Fengyao
authors.initials: F
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, NJU-NJFU Joint Institute of Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing, 210093, People's Republic of China.|Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China.
authors.forename: Ailan
authors.initials: A
authors.lastname: Zou
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, NJU-NJFU Joint Institute of Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing, 210093, People's Republic of China.|Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, NJU-NJFU Joint Institute of Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing, 210093, People's Republic of China.|Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, NJU-NJFU Joint Institute of Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing, 210093, People's Republic of China.|Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China.
authors.forename: Hu
authors.initials: H
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, NJU-NJFU Joint Institute of Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing, 210093, People's Republic of China.|Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China.
authors.forename: Yonghui
authors.initials: Y
authors.lastname: Liao
authors.affiliation: Department of Plant and Microbial Biology, University of California, Berkeley, CA, 94720, USA.
authors.forename: Ren-Jie
authors.initials: RJ
authors.lastname: Tang
authors.affiliation: Jiangsu University of Science and Technology, Zhenjiang, 212003, People's Republic of China.
authors.forename: Tongyi
authors.initials: T
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, NJU-NJFU Joint Institute of Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing, 210093, People's Republic of China.|Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China.
authors.forename: Yanjun
authors.initials: Y
authors.lastname: Pang
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, NJU-NJFU Joint Institute of Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing, 210093, People's Republic of China.|Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, NJU-NJFU Joint Institute of Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing, 210093, People's Republic of China.|Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China.
authors.forename: Rongwu
authors.initials: R
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, NJU-NJFU Joint Institute of Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing, 210093, People's Republic of China. QiJLberkeley@berkeley.edu.|Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China. QiJLberkeley@berkeley.edu.|Department of Plant and Microbial Biology, University of California, Berkeley, CA, 94720, USA. QiJLberkeley@berkeley.edu.
authors.forename: Jinliang
authors.initials: J
authors.lastname: Qi
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, NJU-NJFU Joint Institute of Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing, 210093, People's Republic of China. guihua.lu@nju.edu.cn.|Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China. guihua.lu@nju.edu.cn.
authors.forename: Guihua
authors.initials: G
authors.lastname: Lu
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, NJU-NJFU Joint Institute of Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing, 210093, People's Republic of China. YangYH@nju.edu.cn.|Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China. YangYH@nju.edu.cn.
authors.forename: Yonghua
authors.initials: Y
authors.lastname: Yang
mesh_terms: D003001:Cloning, Molecular; D019295:Computational Biology; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D005030:Ethylenes / Q000494:pharmacology*; D018506:Gene Expression Regulation, Plant / Q000502:physiology*; D029684:Lithospermum / Q000378:metabolism*; D008190:Lyases / Q000235:genetics / Q000378:metabolism*; D009285:Naphthoquinones / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000187:drug effects / Q000378:metabolism*; D030821:Plants, Genetically Modified; D015398:Signal Transduction / Q000502:physiology
chemical_list: D018076:DNA, Complementary; D005030:Ethylenes; D009285:Naphthoquinones; D010940:Plant Proteins; C016101:shikonin; C036216:ethylene; D008190:Lyases; C021773:1-aminocyclopropanecarboxylate synthase
keywords: Ethylene; Hairy root; LeACS-1; Overexpression; RNAi; Shikonin