title: Visualization of plant cell wall lignification using fluorescence-tagged monolignols.
abstract: Lignin is an abundant phenylpropanoid polymer produced by the oxidative polymerization of p-hydroxycinnamyl alcohols (monolignols). Lignification, i.e., deposition of lignin, is a defining feature of secondary cell wall formation in vascular plants, and provides an important mechanism for their disease resistance; however, many aspects of the cell wall lignification process remain unclear partly because of a lack of suitable imaging methods to monitor the process in vivo. In this study, a set of monolignol analogs γ-linked to fluorogenic aminocoumarin and nitrobenzofuran dyes were synthesized and tested as imaging probes to visualize the cell wall lignification process in Arabidopsis thaliana and Pinus radiata under various feeding regimens. In particular, we demonstrate that the fluorescence-tagged monolignol analogs can penetrate into live plant tissues and cells, and appear to be metabolically incorporated into lignifying cell walls in a highly specific manner. The localization of the fluorogenic lignins synthesized during the feeding period can be readily visualized by fluorescence microscopy and is distinguishable from the other wall components such as polysaccharides as well as the pre-existing lignin that was deposited earlier in development.
authors.affiliation: Department of Biochemistry and the US Department of Energy's Great Lakes Bioenergy Research Center (GLBRC), the Wisconsin Energy Institute, University of Wisconsin, 1552 University Avenue, Madison, WI, 53726, USA.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Tobimatsu
authors.forename: Armin
authors.initials: A
authors.lastname: Wagner
authors.forename: Lloyd
authors.initials: L
authors.lastname: Donaldson
authors.forename: Prajakta
authors.initials: P
authors.lastname: Mitra
authors.forename: Claudiu
authors.initials: C
authors.lastname: Niculaes
authors.forename: Oana
authors.initials: O
authors.lastname: Dima
authors.forename: Jeong Im
authors.initials: JI
authors.lastname: Kim
authors.forename: Nickolas
authors.initials: N
authors.lastname: Anderson
authors.forename: Dominique
authors.initials: D
authors.lastname: Loque
authors.forename: Wout
authors.initials: W
authors.lastname: Boerjan
authors.forename: Clint
authors.initials: C
authors.lastname: Chapple
authors.forename: John
authors.initials: J
authors.lastname: Ralph
mesh_terms: D017360:Arabidopsis; D001572:Benzofurans; D002473:Cell Wall / Q000378:metabolism*; D003373:Coumaric Acids; D003374:Coumarins; D005453:Fluorescence; D008031:Lignin / Q000378:metabolism*; D010666:Phenylpropionates / Q000378:metabolism; D028223:Pinus; D059828:Plant Cells / Q000378:metabolism*; D011422:Propionates / Q000378:metabolism; D011523:Protoplasts / Q000378:metabolism; D036226:Seedlings / Q000378:metabolism
chemical_list: D001572:Benzofurans; D003373:Coumaric Acids; D003374:Coumarins; D010666:Phenylpropionates; D011422:Propionates; C036334:2-nitrobenzofuran; C496130:sinapyl alcohol; D008031:Lignin; C495469:p-coumaric acid
keywords: Arabidopsis thaliana; Pinus radiata; cell wall; dehydrogenative polymerization; fluorescence microscopy; fluorescent probes; lignin; technical advance
references.citation: J Struct Biol. 2003 Jul;143(1):77-84
references.pmid: 12892728
references.citation: Plant Cell. 2011 Mar;23(3):1124-37
references.pmid: 21447792
references.citation: ChemSusChem. 2012 Apr;5(4):676-86
references.pmid: 22359379
references.citation: Biotechnol Bioeng. 2009 Sep 1;104(1):68-75
references.pmid: 19489027
references.citation: Plant Physiol. 2010 Jul;153(3):895-905
references.pmid: 20472751
references.citation: Bioconjug Chem. 2003 Jan-Feb;14(1):30-7
references.pmid: 12526689
references.citation: Annu Rev Plant Physiol Plant Mol Biol. 1990;41:455-96
references.pmid: 11543592
references.citation: Science. 2006 Jun 9;312(5779):1491-5
references.pmid: 16627697
references.citation: Planta. 2007 Jun;226(1):267-81
references.pmid: 17333255
references.citation: Annu Rev Plant Biol. 2011;62:273-97
references.pmid: 21370977
references.citation: Biomacromolecules. 2011 May 9;12(5):1752-61
references.pmid: 21410250
references.citation: Phytochemistry. 2001 Jul;57(6):859-73
references.pmid: 11423137
references.citation: Annu Rev Plant Biol. 2003;54:519-46
references.pmid: 14503002
references.citation: Phytochemistry. 2009 Jul-Aug;70(11-12):1366-73
references.pmid: 19712948
references.citation: Cell. 2013 Apr 11;153(2):402-12
references.pmid: 23541512
references.citation: Proc Natl Acad Sci U S A. 1998 Oct 27;95(22):12803-8
references.pmid: 9788995
references.citation: New Phytol. 2010 Jul;187(2):273-285
references.pmid: 20642725
references.citation: Curr Opin Biotechnol. 2008 Apr;19(2):166-72
references.pmid: 18403196
references.citation: Angew Chem Int Ed Engl. 2012 Nov 26;51(48):12005-8
references.pmid: 23109300
references.citation: Biotechnol Bioeng. 2011 Feb;108(2):286-95
references.pmid: 20824689
references.citation: Curr Biol. 2012 Jul 10;22(13):1207-12
references.pmid: 22704988
references.citation: Annu Rev Plant Biol. 2012;63:563-90
references.pmid: 22404466
references.citation: New Phytol. 2012 Dec;196(4):978-1000
references.pmid: 23035778
references.citation: Plant J. 2012 Feb;69(3):542-52
references.pmid: 21978273
references.citation: Plant J. 2005 Jun;42(5):618-40
references.pmid: 15918878
references.citation: Plant Mol Biol. 2006 Dec;62(6):797-807
references.pmid: 17004015
references.citation: Plant Physiol. 2010 Oct;154(2):449-52
references.pmid: 20921161
references.citation: Biomacromolecules. 2005 Sep-Oct;6(5):2688-96
references.pmid: 16153107
references.citation: New Phytol. 2008;180(2):271-295
references.pmid: 19138219
references.citation: Ann Bot. 2005 Apr;95(5):863-8
references.pmid: 15710645
references.citation: Nat Protoc. 2012 Sep;7(9):1694-708
references.pmid: 22918387
references.citation: J Integr Plant Biol. 2010 Feb;52(2):186-94
references.pmid: 20377680
references.citation: Plant Cell. 1992 Nov;4(11):1413-24
references.pmid: 1477555
references.citation: Nat Rev Mol Cell Biol. 2002 Jul;3(7):520-30
references.pmid: 12094218
references.citation: Nature. 2008 Dec 18;456(7224):962-6
references.pmid: 18953331
references.citation: Mol Plant. 2011 Sep;4(5):794-804
references.pmid: 21772029
references.citation: Science. 2012 Nov 23;338(6110):1055-60
references.pmid: 23180856
references.citation: Proc Natl Acad Sci U S A. 1999 Oct 26;96(22):12328-32
references.pmid: 10535921
references.citation: Proc Natl Acad Sci U S A. 2010 Dec 28;107(52):22728-33
references.pmid: 21149736
references.citation: Proc Natl Acad Sci U S A. 2012 Jun 19;109(25):10101-6
references.pmid: 22665765
references.citation: Plant Methods. 2009 Nov 24;5:16
references.pmid: 19930690
references.citation: Annu Rev Genet. 2010;44:337-63
references.pmid: 20809799
references.citation: Phytochemistry. 2010 Oct;71(14-15):1673-83
references.pmid: 20615517
references.citation: Curr Opin Plant Biol. 1999 Apr;2(2):145-52
references.pmid: 10322194
references.citation: Science. 1997 Jul 11;277(5323):235-9
references.pmid: 9211851
references.citation: New Phytol. 2009;184(1):99-113
references.pmid: 19674336
references.citation: Proc Natl Acad Sci U S A. 2012 Jan 24;109(4):1329-34
references.pmid: 22232683
references.citation: Nat Protoc. 2007;2(2):259-62
references.pmid: 17406583
references.citation: J Am Chem Soc. 2005 Jul 6;127(26):9320-1
references.pmid: 15984830
references.citation: Planta. 2013 Mar;237(3):705-15
references.pmid: 23108661
references.citation: Mol Plant. 2012 Mar;5(2):304-17
references.pmid: 22307199
references.citation: Phytochemistry. 2003 Jan;62(2):139-46
references.pmid: 12482448
references.citation: Plant Physiol. 2008 Aug;147(4):1750-60
references.pmid: 18550683
references.citation: J Exp Bot. 2013 Jan;64(1):11-31
references.pmid: 23162114