BPI i norm
ð
Þ ¼
BPI i
P n
i¼0
BPI i
∙100
BPI i(norm) : Base peak intensity of an isotopolog normalized
to the sum of BPI of all isotopologs
DoL i ¼
BPI i norm
ð
Þ ∙i
n
DoL i : Degree of
13
C labeling of an isotopolog with i
13 C
atoms.
n: Number of C atoms in the molecule (40 for carotenoids)
DoL ¼
X n
i¼0
DoL i
DoL: Total degree of
13 C labeling of a metabolite.
Acknowledgments
We thank Ingar Janzik and Andrea Neuwohner for
13 C-labeled leaf
material of peppermint.
References
1. Cazzonelli CI, Pogson BJ (2010) Source to
sink: regulation of carotenoid biosynthesis in
plants. Trends Plant Sci 15:266–274
2. Ruiz-Sola MA ´ , Rodrı ´guez-Conceptio ´ n M
(2012) Carotenoid biosynthesis in Arabidopsis: A colorful pathway. In: The Arabidopsis
Book, vol. 10. American Society of Plant Biologists, p e0158
3. Britton G (1985) Stable isotopes in carotenoid
biochemistry. Pure Appl Chem 57:701–708
4. Lichtenthaler HK (1999) The 1-deoxy-D-xylulose-5-phosphate pathway of isoprenoid biosynthesis in plants. Annu Rev Plant Physiol
Plant Mol Biol 50:47–65
5. Szecowka M, Heise R, Tohge T, NunesNesi A, Vosloh D, Huege J, Feil R, Lunn J,
Nikoloski Z, Stitt M, Fernie AR, Arrivault S
(2013) Metabolic fluxes in an illuminated Arabidopsis rosette. Plant Cell 25:694–714
6. Delwiche CF, Sharkey TD (1993) Rapid
appearance of
13 C in biogenic isoprene when
13
CO 2 is fed to intact leaves. Plant Cell Environ
16:587–591
7. Ghirardo A, Wright LP, Bi Z, Rosenkranz M,
Pulido
P,
Rodrı ´guez-Concepcio ´ n
M,
Niinemetz €
U, Bru ¨ggeman N, Gershenzon J,
Schnitzler J-P (2014) Metabolic flux analysis
of plastidic isoprenoid biosynthesis in poplar
leaves emitting and nonemitting isoprene.
Plant Physiol 165:37–51
8. Beisel KG, Jahnke S, Hofmann D, Ko ¨ppchen S,
Schurr U, Matsubara S (2010) Continuous
turnover of carotenes and chlorophyll a in
mature leaves of Arabidopsis revealed by
14
CO 2 pulse-chase labeling. Plant Physiol
152:2188–2199
9. Aharoni A, de Vos CHR, Verhoeven HA, Maliepaard CA, Kruppa G, Bino R, Goodenowe
DB (2002) Nontargeted metabolome analysis
by use of Fourier transform ion cyclotron mass
spectrometry. OMICS 6:217–234
10. Fiehn O (2002) Metabolomics—the link
between genotypes and phenotypes. Plant
Mol Biol 48:151–171
11. Lei Z, Huhman DV, Sumner LW (2011) Mass
spectrometry strategies in metabolomics. J Biol
Chem 286:25435–25442
12. Heise R, Arrivault S, Szecowka M, Tohge T,
Nunes-Nesi A, Stitt M, Nikoloski Z, Fernie AR
(2014) Flux profiling of photosynthetic carbon
276
Bjo ¨ rn Thiele and Shizue Matsubara
ð
Þ ¼
BPI i
P n
i¼0
BPI i
∙100
BPI i(norm) : Base peak intensity of an isotopolog normalized
to the sum of BPI of all isotopologs
DoL i ¼
BPI i norm
ð
Þ ∙i
n
DoL i : Degree of
13
C labeling of an isotopolog with i
13 C
atoms.
n: Number of C atoms in the molecule (40 for carotenoids)
DoL ¼
X n
i¼0
DoL i
DoL: Total degree of
13 C labeling of a metabolite.
Acknowledgments
We thank Ingar Janzik and Andrea Neuwohner for
13 C-labeled leaf
material of peppermint.
References
1. Cazzonelli CI, Pogson BJ (2010) Source to
sink: regulation of carotenoid biosynthesis in
plants. Trends Plant Sci 15:266–274
2. Ruiz-Sola MA ´ , Rodrı ´guez-Conceptio ´ n M
(2012) Carotenoid biosynthesis in Arabidopsis: A colorful pathway. In: The Arabidopsis
Book, vol. 10. American Society of Plant Biologists, p e0158
3. Britton G (1985) Stable isotopes in carotenoid
biochemistry. Pure Appl Chem 57:701–708
4. Lichtenthaler HK (1999) The 1-deoxy-D-xylulose-5-phosphate pathway of isoprenoid biosynthesis in plants. Annu Rev Plant Physiol
Plant Mol Biol 50:47–65
5. Szecowka M, Heise R, Tohge T, NunesNesi A, Vosloh D, Huege J, Feil R, Lunn J,
Nikoloski Z, Stitt M, Fernie AR, Arrivault S
(2013) Metabolic fluxes in an illuminated Arabidopsis rosette. Plant Cell 25:694–714
6. Delwiche CF, Sharkey TD (1993) Rapid
appearance of
13 C in biogenic isoprene when
13
CO 2 is fed to intact leaves. Plant Cell Environ
16:587–591
7. Ghirardo A, Wright LP, Bi Z, Rosenkranz M,
Pulido
P,
Rodrı ´guez-Concepcio ´ n
M,
Niinemetz €
U, Bru ¨ggeman N, Gershenzon J,
Schnitzler J-P (2014) Metabolic flux analysis
of plastidic isoprenoid biosynthesis in poplar
leaves emitting and nonemitting isoprene.
Plant Physiol 165:37–51
8. Beisel KG, Jahnke S, Hofmann D, Ko ¨ppchen S,
Schurr U, Matsubara S (2010) Continuous
turnover of carotenes and chlorophyll a in
mature leaves of Arabidopsis revealed by
14
CO 2 pulse-chase labeling. Plant Physiol
152:2188–2199
9. Aharoni A, de Vos CHR, Verhoeven HA, Maliepaard CA, Kruppa G, Bino R, Goodenowe
DB (2002) Nontargeted metabolome analysis
by use of Fourier transform ion cyclotron mass
spectrometry. OMICS 6:217–234
10. Fiehn O (2002) Metabolomics—the link
between genotypes and phenotypes. Plant
Mol Biol 48:151–171
11. Lei Z, Huhman DV, Sumner LW (2011) Mass
spectrometry strategies in metabolomics. J Biol
Chem 286:25435–25442
12. Heise R, Arrivault S, Szecowka M, Tohge T,
Nunes-Nesi A, Stitt M, Nikoloski Z, Fernie AR
(2014) Flux profiling of photosynthetic carbon
276
Bjo ¨ rn Thiele and Shizue Matsubara