Metabolic Profiling of Intact Arabidopsis thaliana Leaves during Circadian Cycle Using 1H High Resolution Magic Angle Spinning NMR.
Arabidopsis thaliana is the most widely used model organism for research in plant biology. While significant advances in understanding plant growth and development have been made by focusing on the molecular genetics of Arabidopsis, extracting and understanding the functional framework of metabolism is challenging, both from a technical perspective due to losses and modification during extraction of metabolites from the leaves, and from the biological perspective, due to random variation obscuring how well the function is performed. The purpose of this work is to establish the in vivo metabolic profile directly from the Arabidopsis thaliana leaves without metabolite extraction, to reduce the complexity of the results by multivariate analysis, and to unravel the mitigation of cellular complexity by predominant functional periodicity. To achieve this, we use the circadian cycle that strongly influences metabolic and physiological processes and exerts control over the photosynthetic machinery. High resolution-magic angle spinning nuclear magnetic resonance (HR-MAS NMR) was applied to obtain the metabolic profile directly from intact Arabidopsis leaves. Combining one- and two-dimensional 1H HR-MAS NMR allowed the identification of several metabolites including sugars and amino acids in intact leaves. Multivariate analysis on HR-MAS NMR spectra of leaves throughout the circadian cycle revealed modules of primary metabolites with significant and consistent variations of their molecular components at different time points of the circadian cycle. Since robust photosynthetic performance in plants relies on the functional periodicity of the circadian rhythm, our results show that HR-MAS NMR promises to be an important non-invasive method that can be used for metabolomics of the Arabidopsis thaliana mutants with altered physiology and photosynthetic efficiency.
Leiden Institute of Chemistry, POB 9502, 2300, RA, Leiden, The Netherlands.
D
D
Augustijn
Leiden Institute of Chemistry, POB 9502, 2300, RA, Leiden, The Netherlands.
U
U
Roy
Leiden Institute of Chemistry, POB 9502, 2300, RA, Leiden, The Netherlands.
R
R
van Schadewijk
Leiden Institute of Chemistry, POB 9502, 2300, RA, Leiden, The Netherlands.
H J M
HJ
de Groot
Leiden Institute of Chemistry, POB 9502, 2300, RA, Leiden, The Netherlands.|Institute of Medical Physics and Biophysics, University of Leipzig, Härtelstr. 16-18, D-04107, Leipzig, Germany.
A
A
Alia
Proc Natl Acad Sci U S A. 2001 Aug 14;98(17):10001-5
11481443
Nat Rev Mol Cell Biol. 2006 Mar;7(3):198-210
16496022
Magn Reson Chem. 2014 Aug;52(8):422-9
24842075
Trends Plant Sci. 2000 Dec;5(12):517-22
11120473
Plant Signal Behav. 2014;9(9):e29715
25763705
Food Chem. 2012 Nov 15;135(2):684-93
22868146
Genome Biol. 2006;7(8):R76
16916443
Front Plant Sci. 2012 Mar 14;3:49
22629276
Planta. 2010 Jun;232(1):85-93
20372923
Nature. 2013 Oct 31;502(7473):689-92
24153186
Plant Cell. 1997 Jul;9(7):1235-1244
12237384
Bioessays. 2000 Jan;22(1):38-47
10649289
Front Plant Sci. 2015 Apr 09;6:245
25914715
Curr Metabolomics. 2013;1(1):92-107
26078916
Comput Struct Biotechnol J. 2013 Mar 22;4:e201301009
24688690
Nat Protoc. 2010 Jun;5(6):1019-32
20539278
Wiley Interdiscip Rev Syst Biol Med. 2009 Nov-Dec;1(3):372-9
20228888
Nucleic Acids Res. 2013 Jan;41(Database issue):D801-7
23161693
Magn Reson Chem. 2011 Oct;49(10):659-67
21915899
Arabidopsis Book. 2013;11:e0161
23505340
J Exp Bot. 2002 Apr;53(370):959-70
11912238
Plant Cell. 2005 Dec;17(12):3257-81
16299223
Nucleic Acids Res. 2008 Jan;36(Database issue):D402-8
17984079
Plant J. 2011 Aug;67(3):485-98
21501262
Annu Rev Biophys Biomol Struct. 2004;33:75-93
15139805
Front Plant Sci. 2014 Apr 29;5:163
24808901
Plant Physiol. 2010 Mar;152(3):1251-62
20107023
Methods Mol Biol. 2015;1277:137-46
25677152
Science. 1998 Oct 23;282(5389):662, 679-82
9784120
Magn Reson Chem. 2009 Dec;47 Suppl 1:S147-56
19551810
Environ Sci Technol. 2015 Jun 2;49(11):7056-62
25973714
Front Plant Sci. 2015 Jun 09;6:419
26106401
J Agric Food Chem. 2010 Sep 8;58(17):9675-84
20715777
Anal Chem. 2010 Mar 1;82(5):1643-52
20121204
Plant Physiol. 2007 Nov;145(3):640-52
17885087
J Exp Bot. 2007;58(9):2339-58
17578865
Nat Protoc. 2010 Mar;5(3):536-49
20203669
Sci Rep. 2015 Jul 06;5:11848
26143886
J Agric Food Chem. 2011 Dec 28;59(24):12784-93
22084979
Talanta. 2006 Feb 28;68(5):1683-91
18970515
Plant Physiol. 1987 Feb;83(2):399-407
16665257
Mol Plant. 2014 Jan;7(1):137-55
24121291
Science. 2005 Jul 22;309(5734):630-3
16040710
NMR Biomed. 2000 May;13(3):129-53
10861994
Plant Physiol. 2011 Nov;157(3):1469-82
21949213
Front Bioeng Biotechnol. 2015 Mar 05;3:23
25798438
Planta. 2000 Oct;211(5):743-51
11089689