title: Natural variation in monoterpene synthesis in kiwifruit: transcriptional regulation of terpene synthases by NAC and ETHYLENE-INSENSITIVE3-like transcription factors.
abstract: Two kiwifruit (Actinidia) species with contrasting terpene profiles were compared to understand the regulation of fruit monoterpene production. High rates of terpinolene production in ripe Actinidia arguta fruit were correlated with increasing gene and protein expression of A. arguta terpene synthase1 (AaTPS1) and correlated with an increase in transcript levels of the 2-C-methyl-D-erythritol 4-phosphate pathway enzyme 1-deoxy-D-xylulose-5-phosphate synthase (DXS). Actinidia chinensis terpene synthase1 (AcTPS1) was identified as part of an array of eight tandemly duplicated genes, and AcTPS1 expression and terpene production were observed only at low levels in developing fruit. Transient overexpression of DXS in Nicotiana benthamiana leaves elevated monoterpene synthesis by AaTPS1 more than 100-fold, indicating that DXS is likely to be the key step in regulating 2-C-methyl-D-erythritol 4-phosphate substrate flux in kiwifruit. Comparative promoter analysis identified potential NAC (for no apical meristem [NAM], Arabidopsis transcription activation factor [ATAF], and cup-shaped cotyledon [CUC])-domain transcription factor) and ETHYLENE-INSENSITIVE3-like transcription factor (TF) binding sites in the AaTPS1 promoter, and cloned members of both TF classes were able to activate the AaTPS1 promoter in transient assays. Electrophoretic mobility shift assays showed that AaNAC2, AaNAC3, and AaNAC4 bind a 28-bp fragment of the proximal NAC binding site in the AaTPS1 promoter but not the A. chinensis AcTPS1 promoter, where the NAC binding site was mutated. Activation could be restored by reintroducing multiple repeats of the 12-bp NAC core-binding motif. The absence of NAC transcriptional activation in ripe A. chinensis fruit can account for the low accumulation of AcTPS1 transcript, protein, and monoterpene volatiles in this species. These results indicate the importance of NAC TFs in controlling monoterpene production and other traits in ripening fruits.
authors: The New Zealand Institute for Plant and Food Research Limited, Auckland 1025, New Zealand (N.J.N., X.C., M.Y.W., R.L.P., A.C.A., S.A.G., R.G.A.);School of Biological Sciences, University of Auckland, Auckland 1142, New Zealand (N.J.N., A.C.A.); andThe New Zealand Institute for Plant and Food Research Limited, Palmerston North 4442, New Zealand (A.J.M.) niels.nieuwenhuizen@plantandfood.co.nz.
authors: Niels J
authors: NJ
authors: Nieuwenhuizen
authors: The New Zealand Institute for Plant and Food Research Limited, Auckland 1025, New Zealand (N.J.N., X.C., M.Y.W., R.L.P., A.C.A., S.A.G., R.G.A.);School of Biological Sciences, University of Auckland, Auckland 1142, New Zealand (N.J.N., A.C.A.); andThe New Zealand Institute for Plant and Food Research Limited, Palmerston North 4442, New Zealand (A.J.M.).
authors: Xiuyin
authors: X
authors: Chen
authors: The New Zealand Institute for Plant and Food Research Limited, Auckland 1025, New Zealand (N.J.N., X.C., M.Y.W., R.L.P., A.C.A., S.A.G., R.G.A.);School of Biological Sciences, University of Auckland, Auckland 1142, New Zealand (N.J.N., A.C.A.); andThe New Zealand Institute for Plant and Food Research Limited, Palmerston North 4442, New Zealand (A.J.M.).
authors: Mindy Y
authors: MY
authors: Wang
authors: The New Zealand Institute for Plant and Food Research Limited, Auckland 1025, New Zealand (N.J.N., X.C., M.Y.W., R.L.P., A.C.A., S.A.G., R.G.A.);School of Biological Sciences, University of Auckland, Auckland 1142, New Zealand (N.J.N., A.C.A.); andThe New Zealand Institute for Plant and Food Research Limited, Palmerston North 4442, New Zealand (A.J.M.).
authors: Adam J
authors: AJ
authors: Matich
authors: The New Zealand Institute for Plant and Food Research Limited, Auckland 1025, New Zealand (N.J.N., X.C., M.Y.W., R.L.P., A.C.A., S.A.G., R.G.A.);School of Biological Sciences, University of Auckland, Auckland 1142, New Zealand (N.J.N., A.C.A.); andThe New Zealand Institute for Plant and Food Research Limited, Palmerston North 4442, New Zealand (A.J.M.).
authors: Ramon Lopez
authors: RL
authors: Perez
authors: The New Zealand Institute for Plant and Food Research Limited, Auckland 1025, New Zealand (N.J.N., X.C., M.Y.W., R.L.P., A.C.A., S.A.G., R.G.A.);School of Biological Sciences, University of Auckland, Auckland 1142, New Zealand (N.J.N., A.C.A.); andThe New Zealand Institute for Plant and Food Research Limited, Palmerston North 4442, New Zealand (A.J.M.).
authors: Andrew C
authors: AC
authors: Allan
authors: The New Zealand Institute for Plant and Food Research Limited, Auckland 1025, New Zealand (N.J.N., X.C., M.Y.W., R.L.P., A.C.A., S.A.G., R.G.A.);School of Biological Sciences, University of Auckland, Auckland 1142, New Zealand (N.J.N., A.C.A.); andThe New Zealand Institute for Plant and Food Research Limited, Palmerston North 4442, New Zealand (A.J.M.).
authors: Sol A
authors: SA
authors: Green
authors: The New Zealand Institute for Plant and Food Research Limited, Auckland 1025, New Zealand (N.J.N., X.C., M.Y.W., R.L.P., A.C.A., S.A.G., R.G.A.);School of Biological Sciences, University of Auckland, Auckland 1142, New Zealand (N.J.N., A.C.A.); andThe New Zealand Institute for Plant and Food Research Limited, Palmerston North 4442, New Zealand (A.J.M.).
authors: Ross G
authors: RG
authors: Atkinson
mesh_terms: D029042:Actinidia / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D019883:Alkyl and Aryl Transferases / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D004896:Erythritol / Q000031:analogs & derivatives / Q000378:metabolism; D005030:Ethylenes / Q000378:metabolism; D005638:Fruit / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D039821:Monoterpenes / Q000378:metabolism*; D010802:Phylogeny; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D013403:Sugar Phosphates / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014166:Transferases / Q000235:genetics / Q000378:metabolism
chemical_list: C114232:2-C-methylerythritol 4-phosphate; D005030:Ethylenes; D039821:Monoterpenes; D010940:Plant Proteins; D013403:Sugar Phosphates; D014157:Transcription Factors; C081898:erythritol 4-phosphate; C036216:ethylene; D014166:Transferases; C109461:deoxyxylulose-5-phosphate synthase; D019883:Alkyl and Aryl Transferases; C108877:terpene synthase; D004896:Erythritol
references: Nat Chem Biol. 2008 Oct;4(10):617-23
references: 18776889
references: Theor Appl Genet. 2009 Feb;118(3):541-52
references: 19002427
references: Science. 2002 Apr 12;296(5566):343-6
references: 11951045
references: Plant J. 2005 Feb;41(4):638-49
references: 15686526
references: Appl Microbiol Biotechnol. 2000 Apr;53(4):396-400
references: 10803894
references: Biochemistry. 1998 Sep 1;37(35):12213-20
references: 9724535
references: J Exp Bot. 2012 Mar;63(5):1951-67
references: 22162874
references: Plant Cell Physiol. 2014 Jan;55(1):119-35
references: 24265273
references: Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8915-20
references: 11427737
references: Proc Natl Acad Sci U S A. 2008 Jan 22;105(3):1085-90
references: 18198275
references: Plant Physiol. 2009 May;150(1):365-77
references: 19261734
references: Plant Cell. 2009 Jan;21(1):168-83
references: 19151225
references: Plant J. 2011 Jan;65(1):106-118
references: 21175894
references: Plant Cell. 2005 May;17(5):1612-24
references: 15805488
references: Plant Biotechnol J. 2005 Jan;3(1):17-27
references: 17168896
references: Anal Biochem. 1995 Jul 20;229(1):140-3
references: 8533884
references: Plant Cell. 2008 Feb;20(2):482-94
references: 18296628
references: New Phytol. 2012 Jul;195(2):335-345
references: 22548501
references: Plant Cell. 1996 Jul;8(7):1137-48
references: 8768373
references: Plant Mol Biol. 2003 Oct;53(3):383-97
references: 14750526
references: Nat Commun. 2013;4:2640
references: 24136039
references: Proc Natl Acad Sci U S A. 2001 Nov 20;98(24):13543-8
references: 11698643
references: Plant Cell. 2011 Mar;23(3):923-41
references: 21398570
references: Arch Biochem Biophys. 2000 Apr 15;376(2):354-64
references: 10775423
references: Protein Sci. 1999 May;8(5):978-84
references: 10338008
references: J Chromatogr. 1993 Aug 13;645(1):161-7
references: 23738391
references: Plant J. 2011 Mar;65(6):936-48
references: 21323772
references: Plant Cell Rep. 2007 Nov;26(11):1999-2008
references: 17639404
references: J Mol Biol. 2005 Apr 29;348(2):253-64
references: 15811366
references: Plant Physiol. 2010 Jul;153(3):1280-92
references: 20457803
references: J Biol Chem. 2001 Jun 22;276(25):22901-9
references: 11264287
references: Trends Plant Sci. 2008 Dec;13(12):619-23
references: 18948055
references: Plant Methods. 2005 Dec 18;1:13
references: 16359558
references: Phytochemistry. 2003 Jun;63(3):285-301
references: 12737978
references: Elife. 2013 Jun 11;2:e00675
references: 23795294
references: Plant Physiol. 2008 May;147(1):280-95
references: 18337489
references: Plant Physiol. 2012 Feb;158(2):961-9
references: 22184656
references: J Exp Bot. 2009;60(10):2933-43
references: 19584121
references: Plant Mol Biol. 2006 Nov;62(4-5):683-95
references: 16941216
references: Cell. 2003 Dec 12;115(6):667-77
references: 14675532
references: Plant J. 2008 Jul;55(2):301-10
references: 18397374
references: Theor Appl Genet. 2009 Feb;118(4):653-69
references: 19037624
references: Plant J. 2009 Dec;60(6):1081-95
references: 19891701
references: J Exp Bot. 2012 Sep;63(14):5171-87
references: 22888129
references: Genes Dev. 1998 Dec 1;12(23):3703-14
references: 9851977
references: Plant Cell. 2012 Jun;24(6):2635-48
references: 22669881
references: Plant Cell. 2004 Sep;16(9):2481-98
references: 15319476
references: J Biol Chem. 2013 Jun 7;288(23):16926-16936
references: 23612965
references: New Phytol. 2013 Jan;197(1):36-48
references: 23127167
references: Nature. 2006 Apr 20;440(7087):1078-82
references: 16495946
references: Plant J. 2010 Dec;64(6):936-47
references: 21143675
references: Plant J. 2004 Oct;40(2):188-99
references: 15447646
references: Plant Physiol. 2002 Nov;130(3):1079-89
references: 12427975
references: Plant Physiol. 2006 Nov;142(3):890-900
references: 16980564
references: Methods Enzymol. 2012;515:43-61
references: 22999169
references: Planta. 2010 Oct;232(5):1033-43
references: 20683728
references: Proc Natl Acad Sci U S A. 1998 Apr 14;95(8):4126-33
references: 9539701
references: Plant Cell. 2009 Dec;21(12):4002-17
references: 20028839
references: Phytochemistry. 2003 Apr;62(8):1283-9
references: 12648552
references: Plant Physiol. 2000 Jul;123(3):979-86
references: 10889246
references: Plant J. 2011 Apr;66(1):212-29
references: 21443633
references: Plant Cell. 2009 Oct;21(10):3041-62
references: 19880793
references: Metab Eng. 2013 Nov;20:198-211
references: 24060453
references: Plant Physiol. 2011 Nov;157(3):1568-79
references: 21941001
references: Plant Physiol. 2004 May;135(1):507-15
references: 15133151
references: J Exp Bot. 2012 Oct;63(17):6139-47
references: 23066145
references: Funct Plant Biol. 2007 Oct;34(10):946-961
references: 32689423
references: Cell. 2003 Dec 12;115(6):679-89
references: 14675533
references: DNA Res. 2003 Dec 31;10(6):239-47
references: 15029955
references: J Exp Bot. 2011 Jul;62(11):3821-35
references: 21511911
references: Nature. 2003 Oct 2;425(6957):521-5
references: 14523448
references: Genes Dev. 2000 Dec 1;14(23):3024-36
references: 11114891
references: BMC Genomics. 2008 Jul 27;9:351
references: 18655731
references: Plant J. 2000 Jun;22(6):503-13
references: 10886770
references: Plant Physiol. 2013 Feb;161(2):787-804
references: 23256150
references: Methods Mol Biol. 2000;130:79-83
references: 10589422
references: Anal Biochem. 1987 Nov 1;166(2):368-79
references: 2449095
references: Biochem J. 2010 Feb 09;426(2):183-96
references: 19995345
references: Nat Genet. 2006 Aug;38(8):948-52
references: 16832354
references: J Exp Bot. 2013 May;64(8):2499-510
references: 23599278
references: J Exp Bot. 2002 Oct;53(377):2039-55
references: 12324528