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Regulation of Carotenoid Pigmentation in Corollas of Petunia

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Petunia (Petunia hybrida) is an important ornamental plant, with corolla colors ranging widely from pink to red to purple, owing mainly to anthocyanins. Although there are no bright-yellow-flowered cultivars, some pale-yellow-flowered cultivars accumulate a small amount of carotenoids. To find a key regulatory step that controls carotenoid content in petunia corollas, we compared the expression of carotenoid metabolism genes and carotenoid composition in corollas of white-flowered and pale-yellow-flowered cultivars. Pale yellow corollas tended to have higher expression of biosynthesis genes. The most prominent result was the complete lack of carotenoid cleavage dioxygenase 4a (CCD4a) transcripts in pale yellow corollas. We found two insertions, one in the putative promoter region and the other in the coding region, of the genomic CCD4a sequence of a pale-yellow-flowered cultivar relative to that of a white-flowered cultivar. We consider this the main reason for the lack of CCD4a transcripts. The results suggest that pale yellow corollas have higher carotenoid biosynthesis activity and lower catabolism activity than white corollas. We propose that carotenoid content in petunia corollas is determined by the balance of the degradation and biosynthesis of carotenoids.

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This work was supported by JSPS KAKENHI Grant Number JP25450051.

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Correspondence to Sanae Kishimoto.

Electronic Supplementary Material

Supplementary Fig. S1

Carotenoid analysis in corollas of petunia cultivars. Saponified carotenoid extracts from 0.4 g f.w. of limbs and tubes were analyzed by HPLC. Peaks: 1, (all-E)-neoxanthin; 2, (all-E)-violaxanthin; 3, (9′Z)-neoxanthin; 4, (9Z)-violaxanthin; 5, unknown carotenoid; 6, lutein; 7, antheraxanthin and zeaxanthin; 8, β-carotene. Designations of cultivars are as in Fig. 2. (PPTX 1449 kb)

Supplementary Fig. S2

Carotenoid analysis in leaves of CG and EW. Saponified carotenoid extracts from 0.4 g f.w. of leaves were analyzed by HPLC. Peaks: 2, (all-E)-violaxanthin; 3, (9′Z)-neoxanthin; 4, (9Z)-violaxanthin; 5, unknown carotenoid; 6, lutein; 7, antheraxanthin and zeaxanthin; 8, β-carotene. Designations of cultivars are as in Fig. 2. (PPTX 347 kb)

Supplementary Fig. S3

Alignment of deduced amino acid sequences of CCD1 and CCD4 of various plant species. AtCCD1, from Arabidopsis thaliana (acc. no. NM_116217); AtCCD4 (AtNCED4), A. thaliana (NM_118036); CcCCD4a, Citrus clementina (DQ309330); CcCCD4b, C. clementina (DQ309331); CmCCD4a, Chrysanthemum morifolium (AB247158); CmCCD4b, C. morifolium (AB247160); CsCCD4a, Crocus sativus (EU523662); CsCCD4b, C. sativus (EU523663); PhCCD1, Petunia hybrida (AY576003); PhCCD4a, P. hybrida (LC335775); PhCCD4b, P. hybrida (LC335772). Numbers at branch points indicate bootstrap values (1000 replicates). (PPTX 43 kb)

Supplementary Fig. S4

Genomic sequences of CCD4a presumed promoter and coding regions of California Girl (CG) and Eagle White (EW). Red arrows, genomic PCR primers for insertion detection; green arrows, primers for CCD4a RT-qPCR. Blue bar, insertion; pink bar, deletion in CG relative the sequence from EW. (PPTX 4613 kb)


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Kishimoto, S., Oda-Yamamizo, C. & Ohmiya, A. Regulation of Carotenoid Pigmentation in Corollas of Petunia. Plant Mol Biol Rep 36, 632–642 (2018).

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