11. Use a universal scintillation cocktail for both hydrophilic and
lipophilic samples. A
14 C-quenching correction is recommended for scintillation counting.
Acknowledgments
This work was supported in part by the HarvestPlus research consortium (grant 2014H6320.FRE) to R.W. The authors thank the
former members of the laboratory of Prof. Peter Beyer for their
contributions to establishing this method.
References
1. Beck G, Coman D, Herren E et al (2013)
Characterization of the GGPP synthase gene
family in Arabidopsis thaliana. Plant Mol Biol
82:393–416
2. Ruiz-Sola MA ´ , Coman D, Beck G et al (2016)
Arabidopsis GERANYLGERANYL DIPHOSPHATE SYNTHASE 11 is a hub isozyme
required for the production of most
photosynthesis-related isoprenoids. New Phytol 209:252–264
3. Baranski R, Cazzonelli CI (2016) Carotenoid
biosynthesis and regulation in plants. In: Carotenoids. John Wiley & Sons Ltd, Chichester,
UK, pp 159–189
4. von Lintig J, Welsch R, Bonk M et al (1997)
Light-dependent regulation of carotenoid biosynthesis occurs at the level of phytoene
synthase expression and is mediated by phytochrome in Sinapis alba and Arabidopsis thaliana
seedlings. Plant J 12:625–634
5. Li F, Vallabhaneni R, Wurtzel ET (2008)
PSY3, a new member of the phytoene synthase
gene family conserved in the Poaceae and regulator of abiotic stress-induced root carotenogenesis. Plant Physiol 146:1333–1345
6. Welsch R, Wu ¨st F, B€ ar C et al (2008) A third
phytoene synthase is devoted to abiotic stressinduced abscisic acid formation in rice and
defines functional diversification of phytoene
synthase genes. Plant Physiol 147:367–380
7. Toledo-Ortiz G, Huq E, Rodrı ´guez-Concepcio ´ n M (2010) Direct regulation of phytoene
synthase gene expression and carotenoid biosynthesis by phytochrome-interacting factors.
Proc Natl Acad Sci U S A 107:11626–11631
8. Schledz M, Al-Babili S, von Lintig J et al
(1996) Phytoene synthase from Narcissus
pseudonarcissus: functional expression, galactolipid requirement, topological distribution in
chromoplasts and induction during flowering.
Plant J 10:781–792
9. Giorio G, Stigliani AL, D’Ambrosio C (2008)
Phytoene synthase genes in tomato (Solanum
lycopersicum L.) - new data on the structures,
the deduced amino acid sequences and the
expression patterns. FEBS J 275:527–535
10. Qin X, Coku A, Inoue K, Tian L (2011)
Expression, subcellular localization, and
cis-regulatory structure of duplicated phytoene
synthase genes in melon (Cucumis melo L.).
Planta 234:737–748
11. Farre ´ G, Sanahuja G, Naqvi S et al (2010)
Travel advice on the road to carotenoids in
plants. Plant Sci 179:28–48
12. Maass D, Arango J, Wu ¨st F et al (2009) Carotenoid crystal formation in Arabidopsis and carrot roots caused by increased phytoene
synthase protein levels. PLoS One 4:e6373
13. Schaub P, Rodriguez-Franco M, Cazzonelli CI
et al (2018) Establishment of an Arabidopsis
callus system to study the interrelations of biosynthesis, degradation and accumulation of
carotenoids. PLoS One 13:e0192158
14. Zhou X, Welsch R, Yang Y et al (2015) Arabidopsis OR proteins are the major posttranscriptional regulators of phytoene synthase in
controlling carotenoid biosynthesis. Proc Natl
Acad Sci 112:3558–3563
15. Li L, Yang Y, Xu Q et al (2011) The or gene
enhances carotenoid accumulation and stability
during post-harvest storage of potato tubers.
Mol Plant 5:339–352
16. A ´ lvarez D, Voß B, Maass D et al (2016) Carotenogenesis is regulated by 5’UTR-mediated
translation of Phytoene synthase splice variants.
Plant Physiol 172:2314–2326
17. Welsch R, Zhou X, Yuan H et al (2017) Clp
protease and OR directly control the proteostasis of phytoene synthase, the crucial enzyme
Enzyme Activity of Phytoene Synthase
51
lipophilic samples. A
14 C-quenching correction is recommended for scintillation counting.
Acknowledgments
This work was supported in part by the HarvestPlus research consortium (grant 2014H6320.FRE) to R.W. The authors thank the
former members of the laboratory of Prof. Peter Beyer for their
contributions to establishing this method.
References
1. Beck G, Coman D, Herren E et al (2013)
Characterization of the GGPP synthase gene
family in Arabidopsis thaliana. Plant Mol Biol
82:393–416
2. Ruiz-Sola MA ´ , Coman D, Beck G et al (2016)
Arabidopsis GERANYLGERANYL DIPHOSPHATE SYNTHASE 11 is a hub isozyme
required for the production of most
photosynthesis-related isoprenoids. New Phytol 209:252–264
3. Baranski R, Cazzonelli CI (2016) Carotenoid
biosynthesis and regulation in plants. In: Carotenoids. John Wiley & Sons Ltd, Chichester,
UK, pp 159–189
4. von Lintig J, Welsch R, Bonk M et al (1997)
Light-dependent regulation of carotenoid biosynthesis occurs at the level of phytoene
synthase expression and is mediated by phytochrome in Sinapis alba and Arabidopsis thaliana
seedlings. Plant J 12:625–634
5. Li F, Vallabhaneni R, Wurtzel ET (2008)
PSY3, a new member of the phytoene synthase
gene family conserved in the Poaceae and regulator of abiotic stress-induced root carotenogenesis. Plant Physiol 146:1333–1345
6. Welsch R, Wu ¨st F, B€ ar C et al (2008) A third
phytoene synthase is devoted to abiotic stressinduced abscisic acid formation in rice and
defines functional diversification of phytoene
synthase genes. Plant Physiol 147:367–380
7. Toledo-Ortiz G, Huq E, Rodrı ´guez-Concepcio ´ n M (2010) Direct regulation of phytoene
synthase gene expression and carotenoid biosynthesis by phytochrome-interacting factors.
Proc Natl Acad Sci U S A 107:11626–11631
8. Schledz M, Al-Babili S, von Lintig J et al
(1996) Phytoene synthase from Narcissus
pseudonarcissus: functional expression, galactolipid requirement, topological distribution in
chromoplasts and induction during flowering.
Plant J 10:781–792
9. Giorio G, Stigliani AL, D’Ambrosio C (2008)
Phytoene synthase genes in tomato (Solanum
lycopersicum L.) - new data on the structures,
the deduced amino acid sequences and the
expression patterns. FEBS J 275:527–535
10. Qin X, Coku A, Inoue K, Tian L (2011)
Expression, subcellular localization, and
cis-regulatory structure of duplicated phytoene
synthase genes in melon (Cucumis melo L.).
Planta 234:737–748
11. Farre ´ G, Sanahuja G, Naqvi S et al (2010)
Travel advice on the road to carotenoids in
plants. Plant Sci 179:28–48
12. Maass D, Arango J, Wu ¨st F et al (2009) Carotenoid crystal formation in Arabidopsis and carrot roots caused by increased phytoene
synthase protein levels. PLoS One 4:e6373
13. Schaub P, Rodriguez-Franco M, Cazzonelli CI
et al (2018) Establishment of an Arabidopsis
callus system to study the interrelations of biosynthesis, degradation and accumulation of
carotenoids. PLoS One 13:e0192158
14. Zhou X, Welsch R, Yang Y et al (2015) Arabidopsis OR proteins are the major posttranscriptional regulators of phytoene synthase in
controlling carotenoid biosynthesis. Proc Natl
Acad Sci 112:3558–3563
15. Li L, Yang Y, Xu Q et al (2011) The or gene
enhances carotenoid accumulation and stability
during post-harvest storage of potato tubers.
Mol Plant 5:339–352
16. A ´ lvarez D, Voß B, Maass D et al (2016) Carotenogenesis is regulated by 5’UTR-mediated
translation of Phytoene synthase splice variants.
Plant Physiol 172:2314–2326
17. Welsch R, Zhou X, Yuan H et al (2017) Clp
protease and OR directly control the proteostasis of phytoene synthase, the crucial enzyme
Enzyme Activity of Phytoene Synthase
51