Abstract
The blue colour of Vanda coerulea petals is slowly produced during flower development but quickly disappears when the flowers are pollinated. To investigate the molecular basis of the phenomenon, we isolated a novel cytochrome P450 gene, CYP76AB1, from this plant by polymerase chain reaction (PCR) based on the conserved regions of flavonoid 3′,5′-hydroxylase amino acid sequences. CYP76AB1 transcripts were detectable at low level in the late phase of flower development when the petals were light blue, but became abundant in the subsequent opening flower stage, as the petals turned blue. When the flowers were pollinated they turned white, and the CYP76AB1 transcripts returned to low level. This indicates that the expression of CYP76AB1 may regulate petal colour of V. coerulea, and be a target for developing permanently blue flowers.
Abbreviations
- cDNA:
-
complementary deoxyribonucleic acid
- CYP75A:
-
cytochrome P450, family 75, subfamily A
- CYP76AB1:
-
cytochrome P450, family 76, subfamily AB, isoform 1
- CYP93B:
-
cytochrome P450, family 93, subfamily B
- dNTP:
-
deoxynucleotide triphosphate
- F3′H:
-
flavonoid 3′-hydroxylase
- F3′5′H:
-
flavonoid 3′,5′-hydroxylase
- FNSII:
-
flavone synthase II
- gDNA:
-
genomic deoxyribonucleic acid
- P450s:
-
cytochromes P450
- ORF:
-
open reading frame
- RACE:
-
rapid amplification of cDNA end
- RT-PCR:
-
reverse transcriptase polymerase chain reaction
References
Akashi, T., Fukuchi-Mizutani, M., Aoki, T., Ueyama, Y., Yonekura-Sakakibara, K., Tanaka, Y., Kusumi, T., Ayabe, S.: Molecular cloning and biochemical characterization of a novel cytochrome P450, flavone synthase II, that catalyzes direct conversion of flavanones to flavones. — Plant Cell Physiol. 40: 1182–1186, 1999.
Dellaporta, S., Wood, J., Hicks, J.: A plant DNA minipreparation: version II. — Plant mol. Biol. Rep. 1: 19–21, 1983.
Feria-Romero, I., Astudillo-de la Vega, H., Chavez-Soto, M.A., Rivera-Arce, E., López, M., Serrano, H., Lozoya, X.: RAPD markers associated with quercetin accumulation in Psidium guajava. — Biol. Plant. 53: 125–128, 2009.
Holton, T.A., Brugliera, F., Lester, D.R., Tanaka, Y., Hyland, C.D., Menting, J.G., Lu, C.Y., Farcy, E., Stevenson, T.W., Cornish, E.C.: Cloning and expression of cytochrome P450 genes controlling flower colour. — Nature 366: 276–279, 1993.
Imaishi, H., Ishitobi, U.: Molecular cloning of CYP76A3, a novel cytochrome P450 from Petunia hybrida catalyzing the ω-hydroxylation of myristic acid. — Biol. Plant. 52: 242–250, 2008.
Imaishi, H., Petkova-Andonova, M.: Molecular cloning of CYP76B9, a cytochrome P450 from Petunia hybrida, catalyzing the ω-hydroxylation of capric acid and lauric acid. — Biosci. Biotechnol. Biochem. 71: 104–113, 2007.
Martens, S., Forkmann, G.: Cloning and expression of flavone synthase II from Gerbera hybrids. — Plant J. 20: 611–618, 1999.
Martens, S., Mithofer, A.: Flavones and flavone synthases. — Phytochemistry 66: 2399–2407, 2005.
Mizutani, M., Ward, E., Ohta, D.: Cytochrome P450 superfamily in Arabidopsis thaliana: isolation of cDNAs, differential expression, and RFLP mapping of multiple cytochromes P450. — Plant mol. Biol. 37: 39–52, 1998.
Mori, S., Kobayashi, H., Hoshi, Y., Kondo, M., Nakano, M.: Heterologous expression of the flavonoid 3′,5′-hydroxylase gene of Vinca major alters flower color in transgenic Petunia hybrida. — Plant Cell Rep. 22: 415–421, 2004.
Nielsen, K.M., Podivinsky, E.: cDNA cloning and endogenous expression of a flavonoid 3′,5′-hydroxylase from petals of lisianthus (Eustoma grandiflorum). — Plant Sci. 129: 167–174, 1997.
Su, V., Hsu, B.D.: Cloning and expression of a putative cytochrome P450 gene that influences the colour of Phalaenopsis flowers. — Biotechnol. Lett. 25: 1933–1939, 2003.
Verwoerd, T.C., Dekker, B.M., Hoekema, A.: A small-scale procedure for the rapid isolation of plant RNAs. — Nucl. Acids Res. 17: 2362, 1989.
Wang, J., Ming, F., Han, Y., Shen, D.: Flavonoid-3′,5′- hydroxylase from Phalaenopsis: a novel member of cytochrome P450s, its cDNA cloning, endogenous expression and molecular modeling. — Biotechnol. Lett. 28: 327–334, 2006.
Yukiko, U., Yukihisa, K., Yuko, F., Masako, F.M., Hideo, O., Takaaki, K., Takashi, I., Yoshikazu, T.: Molecular characterization of the flavonoid biosynthetic pathway and flower color modification of Nierembergia sp. — Plant Biotechnol. 23: 19–24, 2006.
Acknowledgements
This work was supported by Coordinating Center for Thai Government Science and Technology Scholarship Students, National Science and Technology Development Agency, Ministry of Science and Technology, Thailand (F-31-312-79-01).
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Ratanasut, K., Wongkhamprai, B. & Maknoi, S. Expression of a CYP76AB1 correlates with the sequential white-blue-white colour transition of Vanda coerulea petals. Biol Plant 55, 353–356 (2011). https://doi.org/10.1007/s10535-011-0053-3
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DOI: https://doi.org/10.1007/s10535-011-0053-3