Fig 1.
Xanthophyll biosynthesis pathway in plants.
Abbreviations: CYP97C, heme-containing cytochrome P450 carotene ε-ring hydroxylase; HYDB, β-carotene hydroxylase (non-heme di-iron β-carotene hydroxylase (BCH) and heme-containing cytochrome P450 β-ring hydroxylases CYP97A and CYP97B); LYCB, lycopene β-cyclase; LYCE, lycopene ε-cyclase; VDE, violaxanthin de-epoxidase; ZEP, zeaxanthin epoxidase.
Fig 2.
Multiple alignments of CYP97C protein sequences from maize (Zm, Zea mays; GenBank: GU130217), rice (Os, Oryza sativa; GenBank: AK065689), Arabidopsis (At, Arabidopsis thaliana; GenBank: NM_115173) and tomato (Sl, Solanum lycopersicon; GenBank: EU849604).
Fig 3.
Gene structures for maize, rice, tomato and A. thaliana CYP97C homologs.
Orange boxes represent exons. Blue lines represent introns. The lengths of DNA sequences are indicated on the right. Zm, Zea mays cDNA; GenBank: GU130217), rice (Os, Oryza sativa cDNA; GenBank: AK065689), Arabidopsis (At, Arabidopsis thaliana cDNA; GenBank: NM_115173), tomato (Sl, Solanum lycopersicon cDNA and genomic DNA; GenBank: EU849604 and EU849603).
Fig 4.
DNA blot analysis of ZmCYP97C19 transgene in A. thaliana wild type, lut1 mutant and three different transgenic lines transformed with ZmCYP97C19 driven by the CaMV 35S promoter.
Genomic DNA (10 μg) from rosette leaves was separately digested with EcoRI (A) and XbaI (B). The DNA blot was hybridized with a ZmCYP97C19 gene probe. WT, wild-type (Col-0); Lut1, lut1 mutant; L1, line 1; L2, line 2; L3, line 3.
Fig 5.
RNA blot analysis of maize CYP97C19 transgene expression in A. thaliana wild-type, lut1 mutant and three different transgenic lines transformed with ZmCYP97C19 driven by the CaMV 35S promoter.
Each lane was loaded with 20 μg total RNA from rosette leaves. Ribosomal RNA stained with ethidium bromide is shown as a loading control. The blot was hybridized with a ZmCYP97C19 probe. WT, wild type (Col-0); Lut1, lut1 mutant; L1, line 1; L2, line 2; L3, line 3.
Table 1.
Abundance of individual carotenoids in Arabidopsis thaliana leaves (%) and the total carotenoid content (μg/g dry weight).
Fig 6.
HPLC analysis of carotenoids in rosette leaves of A. thaliana wild-type, lut1 mutant and transgenic lines transformed with ZmCYP97C19 driven by the CaMV 35S promoter.
Peak 1, violaxanthin; peak 2, lutein, peak 3, β-carotene; peak 3', β-carotene cis isomer; peak 4, antheraxanthin; peak 5, zeaxanthin; peak 6, zeinoxanthin; peak 7, α-carotene. Wild type, Col-0; Lut1, lut1 mutant; CYP97C in Lut 1, transgenic lines.