title: Regulation of Nicotine Biosynthesis by an Endogenous Target Mimicry of MicroRNA in Tobacco.
abstract: The interaction between noncoding endogenous target mimicry (eTM) and its corresponding microRNA (miRNA) is a newly discovered regulatory mechanism and plays pivotal roles in various biological processes in plants. Tobacco (Nicotiana tabacum) is a model plant for studying secondary metabolite alkaloids, of which nicotine accounts for approximately 90%. In this work, we identified four unique tobacco-specific miRNAs that were predicted to target key genes of the nicotine biosynthesis and catabolism pathways and an eTM, novel tobacco miRNA (nta)-eTMX27, for nta-miRX27 that targets QUINOLINATE PHOSPHORIBOSYLTRANSFERASE2 (QPT2) encoding a quinolinate phosphoribosyltransferase. The expression level of nta-miRX27 was significantly down-regulated, while that of QPT2 and nta-eTMX27 was significantly up-regulated after topping, and consequently, nicotine content increased in the topping-treated plants. The topping-induced down-regulation of nta-miRX27 and up-regulation of QPT2 were only observed in plants with a functional nta-eTMX27 but not in transgenic plants containing an RNA interference construct targeting nta-eTMX27. Our results demonstrated that enhanced nicotine biosynthesis in the topping-treated tobacco plants is achieved by nta-eTMX27-mediated inhibition of the expression and functions of nta-miRX27. To our knowledge, this is the first report about regulation of secondary metabolite biosynthesis by an miRNA-eTM regulatory module in plants.
authors.affiliation: Institute of Crop Science and Research Center for Air Pollution and Health (F.L., W.W., N.Z., C.Y., E.S., J.Q., L.F.) and Institute of Biotechnology (F.L., N.Z., X.Z.), Zhejiang University, Hangzhou 310058, China;Yunnan Academy of Tobacco Agricultural Sciences and China Tobacco Breeding Research Center at Yunnan, Yuxi 653100, China (B.X., X.W.);National Center of Tobacco Genes, Zhengzhou 450001, China (P.C.);Commonwealth Scientific and Industrial Research Organization Agriculture Flagship, Canberra, Australian Capital Territory 2601, Australia (Q.-H.Z.); andDepartment of Pharmaceutical Sciences, North Carolina Central University, Durham, North Carolina 27707 (J.X.).
authors.forename: Fangfang
authors.initials: F
authors.lastname: Li
authors.affiliation: Institute of Crop Science and Research Center for Air Pollution and Health (F.L., W.W., N.Z., C.Y., E.S., J.Q., L.F.) and Institute of Biotechnology (F.L., N.Z., X.Z.), Zhejiang University, Hangzhou 310058, China;Yunnan Academy of Tobacco Agricultural Sciences and China Tobacco Breeding Research Center at Yunnan, Yuxi 653100, China (B.X., X.W.);National Center of Tobacco Genes, Zhengzhou 450001, China (P.C.);Commonwealth Scientific and Industrial Research Organization Agriculture Flagship, Canberra, Australian Capital Territory 2601, Australia (Q.-H.Z.); andDepartment of Pharmaceutical Sciences, North Carolina Central University, Durham, North Carolina 27707 (J.X.).
authors.forename: Weidi
authors.initials: W
authors.lastname: Wang
authors.affiliation: Institute of Crop Science and Research Center for Air Pollution and Health (F.L., W.W., N.Z., C.Y., E.S., J.Q., L.F.) and Institute of Biotechnology (F.L., N.Z., X.Z.), Zhejiang University, Hangzhou 310058, China;Yunnan Academy of Tobacco Agricultural Sciences and China Tobacco Breeding Research Center at Yunnan, Yuxi 653100, China (B.X., X.W.);National Center of Tobacco Genes, Zhengzhou 450001, China (P.C.);Commonwealth Scientific and Industrial Research Organization Agriculture Flagship, Canberra, Australian Capital Territory 2601, Australia (Q.-H.Z.); andDepartment of Pharmaceutical Sciences, North Carolina Central University, Durham, North Carolina 27707 (J.X.).
authors.forename: Nan
authors.initials: N
authors.lastname: Zhao
authors.affiliation: Institute of Crop Science and Research Center for Air Pollution and Health (F.L., W.W., N.Z., C.Y., E.S., J.Q., L.F.) and Institute of Biotechnology (F.L., N.Z., X.Z.), Zhejiang University, Hangzhou 310058, China;Yunnan Academy of Tobacco Agricultural Sciences and China Tobacco Breeding Research Center at Yunnan, Yuxi 653100, China (B.X., X.W.);National Center of Tobacco Genes, Zhengzhou 450001, China (P.C.);Commonwealth Scientific and Industrial Research Organization Agriculture Flagship, Canberra, Australian Capital Territory 2601, Australia (Q.-H.Z.); andDepartment of Pharmaceutical Sciences, North Carolina Central University, Durham, North Carolina 27707 (J.X.).
authors.forename: Bingguang
authors.identifier: http://orcid.org/0000-0002-1791-4461
authors.initials: B
authors.lastname: Xiao
authors.affiliation: Institute of Crop Science and Research Center for Air Pollution and Health (F.L., W.W., N.Z., C.Y., E.S., J.Q., L.F.) and Institute of Biotechnology (F.L., N.Z., X.Z.), Zhejiang University, Hangzhou 310058, China;Yunnan Academy of Tobacco Agricultural Sciences and China Tobacco Breeding Research Center at Yunnan, Yuxi 653100, China (B.X., X.W.);National Center of Tobacco Genes, Zhengzhou 450001, China (P.C.);Commonwealth Scientific and Industrial Research Organization Agriculture Flagship, Canberra, Australian Capital Territory 2601, Australia (Q.-H.Z.); andDepartment of Pharmaceutical Sciences, North Carolina Central University, Durham, North Carolina 27707 (J.X.).
authors.forename: Peijian
authors.identifier: http://orcid.org/0000-0001-9991-423X
authors.initials: P
authors.lastname: Cao
authors.affiliation: Institute of Crop Science and Research Center for Air Pollution and Health (F.L., W.W., N.Z., C.Y., E.S., J.Q., L.F.) and Institute of Biotechnology (F.L., N.Z., X.Z.), Zhejiang University, Hangzhou 310058, China;Yunnan Academy of Tobacco Agricultural Sciences and China Tobacco Breeding Research Center at Yunnan, Yuxi 653100, China (B.X., X.W.);National Center of Tobacco Genes, Zhengzhou 450001, China (P.C.);Commonwealth Scientific and Industrial Research Organization Agriculture Flagship, Canberra, Australian Capital Territory 2601, Australia (Q.-H.Z.); andDepartment of Pharmaceutical Sciences, North Carolina Central University, Durham, North Carolina 27707 (J.X.).
authors.forename: Xingfu
authors.initials: X
authors.lastname: Wu
authors.affiliation: Institute of Crop Science and Research Center for Air Pollution and Health (F.L., W.W., N.Z., C.Y., E.S., J.Q., L.F.) and Institute of Biotechnology (F.L., N.Z., X.Z.), Zhejiang University, Hangzhou 310058, China;Yunnan Academy of Tobacco Agricultural Sciences and China Tobacco Breeding Research Center at Yunnan, Yuxi 653100, China (B.X., X.W.);National Center of Tobacco Genes, Zhengzhou 450001, China (P.C.);Commonwealth Scientific and Industrial Research Organization Agriculture Flagship, Canberra, Australian Capital Territory 2601, Australia (Q.-H.Z.); andDepartment of Pharmaceutical Sciences, North Carolina Central University, Durham, North Carolina 27707 (J.X.).
authors.forename: Chuyu
authors.identifier: http://orcid.org/0000-0003-0903-0356
authors.initials: C
authors.lastname: Ye
authors.affiliation: Institute of Crop Science and Research Center for Air Pollution and Health (F.L., W.W., N.Z., C.Y., E.S., J.Q., L.F.) and Institute of Biotechnology (F.L., N.Z., X.Z.), Zhejiang University, Hangzhou 310058, China;Yunnan Academy of Tobacco Agricultural Sciences and China Tobacco Breeding Research Center at Yunnan, Yuxi 653100, China (B.X., X.W.);National Center of Tobacco Genes, Zhengzhou 450001, China (P.C.);Commonwealth Scientific and Industrial Research Organization Agriculture Flagship, Canberra, Australian Capital Territory 2601, Australia (Q.-H.Z.); andDepartment of Pharmaceutical Sciences, North Carolina Central University, Durham, North Carolina 27707 (J.X.).
authors.forename: Enhui
authors.initials: E
authors.lastname: Shen
authors.affiliation: Institute of Crop Science and Research Center for Air Pollution and Health (F.L., W.W., N.Z., C.Y., E.S., J.Q., L.F.) and Institute of Biotechnology (F.L., N.Z., X.Z.), Zhejiang University, Hangzhou 310058, China;Yunnan Academy of Tobacco Agricultural Sciences and China Tobacco Breeding Research Center at Yunnan, Yuxi 653100, China (B.X., X.W.);National Center of Tobacco Genes, Zhengzhou 450001, China (P.C.);Commonwealth Scientific and Industrial Research Organization Agriculture Flagship, Canberra, Australian Capital Territory 2601, Australia (Q.-H.Z.); andDepartment of Pharmaceutical Sciences, North Carolina Central University, Durham, North Carolina 27707 (J.X.).
authors.forename: Jie
authors.initials: J
authors.lastname: Qiu
authors.affiliation: Institute of Crop Science and Research Center for Air Pollution and Health (F.L., W.W., N.Z., C.Y., E.S., J.Q., L.F.) and Institute of Biotechnology (F.L., N.Z., X.Z.), Zhejiang University, Hangzhou 310058, China;Yunnan Academy of Tobacco Agricultural Sciences and China Tobacco Breeding Research Center at Yunnan, Yuxi 653100, China (B.X., X.W.);National Center of Tobacco Genes, Zhengzhou 450001, China (P.C.);Commonwealth Scientific and Industrial Research Organization Agriculture Flagship, Canberra, Australian Capital Territory 2601, Australia (Q.-H.Z.); andDepartment of Pharmaceutical Sciences, North Carolina Central University, Durham, North Carolina 27707 (J.X.).
authors.forename: Qian-Hao
authors.identifier: http://orcid.org/0000-0002-6505-7417
authors.initials: QH
authors.lastname: Zhu
authors.affiliation: Institute of Crop Science and Research Center for Air Pollution and Health (F.L., W.W., N.Z., C.Y., E.S., J.Q., L.F.) and Institute of Biotechnology (F.L., N.Z., X.Z.), Zhejiang University, Hangzhou 310058, China;Yunnan Academy of Tobacco Agricultural Sciences and China Tobacco Breeding Research Center at Yunnan, Yuxi 653100, China (B.X., X.W.);National Center of Tobacco Genes, Zhengzhou 450001, China (P.C.);Commonwealth Scientific and Industrial Research Organization Agriculture Flagship, Canberra, Australian Capital Territory 2601, Australia (Q.-H.Z.); andDepartment of Pharmaceutical Sciences, North Carolina Central University, Durham, North Carolina 27707 (J.X.).
authors.forename: Jiahua
authors.initials: J
authors.lastname: Xie
authors.affiliation: Institute of Crop Science and Research Center for Air Pollution and Health (F.L., W.W., N.Z., C.Y., E.S., J.Q., L.F.) and Institute of Biotechnology (F.L., N.Z., X.Z.), Zhejiang University, Hangzhou 310058, China;Yunnan Academy of Tobacco Agricultural Sciences and China Tobacco Breeding Research Center at Yunnan, Yuxi 653100, China (B.X., X.W.);National Center of Tobacco Genes, Zhengzhou 450001, China (P.C.);Commonwealth Scientific and Industrial Research Organization Agriculture Flagship, Canberra, Australian Capital Territory 2601, Australia (Q.-H.Z.); andDepartment of Pharmaceutical Sciences, North Carolina Central University, Durham, North Carolina 27707 (J.X.) fanlj@zju.edu.cn zzhou@zju.edu.cn.
authors.forename: Xueping
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Institute of Crop Science and Research Center for Air Pollution and Health (F.L., W.W., N.Z., C.Y., E.S., J.Q., L.F.) and Institute of Biotechnology (F.L., N.Z., X.Z.), Zhejiang University, Hangzhou 310058, China;Yunnan Academy of Tobacco Agricultural Sciences and China Tobacco Breeding Research Center at Yunnan, Yuxi 653100, China (B.X., X.W.);National Center of Tobacco Genes, Zhengzhou 450001, China (P.C.);Commonwealth Scientific and Industrial Research Organization Agriculture Flagship, Canberra, Australian Capital Territory 2601, Australia (Q.-H.Z.); andDepartment of Pharmaceutical Sciences, North Carolina Central University, Durham, North Carolina 27707 (J.X.) fanlj@zju.edu.cn zzhou@zju.edu.cn.
authors.forename: Longjiang
authors.identifier: http://orcid.org/0000-0001-7695-7958
authors.initials: L
authors.lastname: Fan
mesh_terms: D019143:Evolution, Molecular; D018506:Gene Expression Regulation, Plant; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D018716:Molecular Mimicry; D009538:Nicotine / Q000096:biosynthesis* / Q000235:genetics; D010430:Pentosyltransferases / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism*; D000098553:Nicotinate-Nucleotide Diphosphorylase (Carboxylating)
chemical_list: D035683:MicroRNAs; D010940:Plant Proteins; D009538:Nicotine; D010430:Pentosyltransferases; D000098553:Nicotinate-Nucleotide Diphosphorylase (Carboxylating)