Isolation and Functional Characterization of a Lycopene β-cyclase Gene Promoter from Citrus.
Lycopene β-cyclases are key enzymes located at the branch point of the carotenoid biosynthesis pathway. However, the transcriptional regulatory mechanisms of LCYb1 in citrus with abundant carotenoid accumulation are still unclear. To understand the molecular basis of CsLCYb1 expression, we isolated and functionally characterized the 5' upstream sequences of CsLCYb1 from citrus. The full-length CsLCYb1 promoter and a series of its 5' deletions were fused to the β-glucuronidase (GUS) reporter gene and transferred into different plants (tomato, Arabidopsis and citrus callus) to test the promoter activities. The results of all transgenic species showed that the 1584 bp upstream region from the translational start site displayed maximal promoter activity, and the minimal promoter containing 746 bp upstream sequences was sufficient for strong basal promoter activity. Furthermore, the CsLCYb1 promoter activity was developmentally and tissue-specially regulated in transgenic Arabidopsis, and it was affected by multiple hormones and environmental cues in transgenic citrus callus under various treatments. Finer deletion analysis identified an enhancer element existing as a tandem repeat in the promoter region between -574 to -513 bp and conferring strong promoter activity. The copy numbers of the enhancer element differed among various citrus species, leading to the development of a derived simple sequence repeat marker to distinguish different species. In conclusion, this study elucidates the expression characteristics of the LCYb1 promoter from citrus and further identifies a novel enhancer element required for the promoter activity. The characterized promoter fragment would be an ideal candidate for genetic engineering and seeking of upstream trans-acting elements.
Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University Wuhan, China.
Suwen
S
Lu
Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University Wuhan, China.
Yin
Y
Zhang
Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University Wuhan, China.
Xiongjie
X
Zheng
Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University Wuhan, China.
Kaijie
K
Zhu
Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University Wuhan, China.
Qiang
Q
Xu
Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University Wuhan, China.
Xiuxin
X
Deng
carotenoid; cis-element; citrus; enhancer; lycopene β-cyclase; promoter
Plant Cell. 1996 Sep;8(9):1613-26
8837512
BMC Genomics. 2009 Nov 18;10:540
19922663
Front Plant Sci. 2013 Jun 14;4:198
23785378
Plant J. 1996 Apr;9(4):505-12
8624513
Trends Plant Sci. 1999 Jun;4(6):210-214
10366876
BMC Plant Biol. 2010 Apr 09;10:61
20380705
Plant J. 1998 Dec;16(6):735-43
10069079
Plant J. 2011 Sep;67(5):917-28
21585571
Methods Enzymol. 1993;214:256-69
8469143
Ann Bot. 2013 Jan;111(1):1-19
23104641
Plant Physiol. 1987 Nov;85(3):726-32
16665768
Science. 2012 Jun 29;336(6089):1711-5
22745430
Genes Dev. 1996 Mar 15;10(6):711-24
8598298
J Exp Bot. 2012 Jan;63(2):871-86
21994171
Plant Physiol. 2006 Jan;140(1):3-11
16403736
Transgenic Res. 2012 Oct;21(5):1043-56
22297392
Plant Cell. 2013 Feb;25(2):371-86
23386264
PLoS Genet. 2010 Sep 09;6(9):e1001102
20838596
Nucleic Acids Res. 2002 Jan 1;30(1):325-7
11752327
BMC Genomics. 2012 Jan 10;13:10
22230690
Trends Plant Sci. 2004 Aug;9(8):385-90
15358269
Plant Cell. 1993 Apr;5(4):379-87
8485401
Plant Physiol. 2004 Aug;135(4):2150-61
15310827
Plant J. 2002 Jan;29(1):1-10
12060222
EMBO J. 1987 Dec 20;6(13):3901-7
3327686
Prog Lipid Res. 2004 May;43(3):228-65
15003396
Annu Rev Plant Biol. 2006;57:711-38
16669779
J Exp Bot. 2012 Jul;63(12):4403-17
22611233
J Agric Food Chem. 2006 Jul 26;54(15):5474-81
16848534
Planta. 2003 Jan;216(3):523-34
12520345
Mol Plant. 2015 Jan;8(1):68-82
25578273
Genomics. 2010 Oct;96(4):239-50
20633636
Proc Natl Acad Sci U S A. 2012 Apr 3;109(14):5535-40
22431637
Plant Physiol. 2001 Apr;125(4):1558-66
11299337
Planta. 2012 Oct;236(4):1315-25
22729824
Plant Cell. 1995 Jul;7(7):1027-38
7640523
Nat Genet. 2013 Jan;45(1):59-66
23179022
J Exp Bot. 2009;60(6):1783-97
19325166
J Plant Physiol. 2013 Sep 15;170(14):1295-9
23643306
BMC Plant Biol. 2015 May 08;15:114
25953041
Science. 2002 Apr 12;296(5566):343-6
11951045
Mol Plant Pathol. 2008 Mar;9(2):251-68
18705856
Plant Physiol. 2013 Mar;161(3):1476-85
23292788
Mol Plant. 2015 Jan;8(1):28-39
25578270
Photochem Photobiol. 1996 Mar;63(3):257-264
8881328
J Biol Chem. 1998 Nov 13;273(46):30651-9
9804838
Plant Cell Rep. 2011 Jun;30(6):973-80
21243360
Proc Natl Acad Sci U S A. 2010 Jun 22;107(25):11626-31
20534526
Plant Cell Environ. 2014 Feb;37(2):273-89
23777240
Mol Aspects Med. 2005 Dec;26(6):459-516
16309738
Physiol Plant. 2011 Jan;141(1):1-10
20875059
Plant Physiol. 2004 Feb;134(2):824-37
14739348
Plant Physiol. 2007 Nov;145(3):1073-85
17873090
Nucleic Acids Res. 1999 Jan 1;27(1):297-300
9847208
J Biol Chem. 2002 Mar 29;277(13):11582-90
11782472
Science. 2012 Mar 16;335(6074):1348-51
22422982
J Agric Food Chem. 2012 Jan 11;60(1):197-201
22026557
J Agric Food Chem. 2006 Jun 14;54(12):4397-406
16756373