Figure 1.
Simplified flavonoid pathway demonstrating the influence of DFR substrate acceptance on the establishment of flower colour.
Figure 2.
Multiple alignment of the deduced amino acid sequences encoded by DFRs isolated from F. vesca cv Alexandria during early (KC894042, KC894049) and late (KC894043, KC894044, KC894050, KC894051) stages, cv. Red Wonder early (KC894045, KC894052) and late (KC894046, KC894053) stages and F.
×ananassa cv. Elsanta early (KC894047, KC894054) and late (KC894048, KC894055) stages. abbreviations: RW: Red Wonder, Alex: Alexandria, ES: Elsanta, es: early stage, ls: late stage.
Table 1.
DFR cDNA clones isolated from Fragaria species.
Figure 3.
Neighbour-joining tree of DFR amino acid sequences of Fragaria and various species published in the NCBI GenBank.
DFR1 F. vesca cv Alexandria, early stage (KC894042), late stage 1 (KC894043), late stage 2 (KC894044), cv Red Wonder early stage (KC894045), late stage (KC894046), F.×ananassa cv. Elsanta, early stage (KC894047), late stage (KC894048), DFR2 F. vesca cv Alexandria, early stage (KC894049), late stage 1 (KC894050), late stage 2 (KC894051), cv Red Wonder early stage (KC894052), late stage (KC894053), F.×ananassa cv. Elsanta, early stage (KC894054), late stage (KC894055), Angelonia×angustifolia (KF285561), Camellia sinensis (AAT84073), Citrus sinensis (AAS00611), Crataegus monogynae (AAX16491), Delphinium belladonna (BAF49325), Garcinia mangostane (ACM62744), Gerbera hybrida (CAA78930), Malus×domestica (AAO39816, AAO39817), Medicago truncatula (AAR27014, AAR27015), Onobrychis viciifolia (AEF14420), Petunia×hybrida (AAF60298), Pyrus communis (AAO39818), Rosa hybrida (AAX1242), Triticum aestivum (AAQ77347), Vitis vinifera (AAX12423, XP002275531), Vaccinium macrocarpon (AAL89715, AAL8971, AAX12420). The bootstrap values are indicated next to the relevant nodes (1000 replicates).
Table 2.
Characterization of recombinant DFR1 and DFR2 from Fragaria×ananassa cv. Elsanta obtained from heterologous expression of DFRs (KC894048, KC894055) in yeast.
Figure 4.
Quantitative expression of DFR1 and DFR2 normalized to glyceraldehyde 3-phosphate dehydrogenase in receptacle and achenes of Fragaria fruits during the different stages of fruit development.
a: F.×ananassa receptacle, b: F.×ananassa achenes, c: F. vesca receptacle, d: F. vesca achenes. red: DFR1, grey: DFR2. Data were calculated from three biological replicates with at least two technical replicates for each and with error bars representing the standard deviation.
Figure 5.
Specific DFR activity [nkat/kg protein] with DHQ and DHK as substrates in receptacle and achenes throughout the different stages of fruit development and ripening in F.×ananassa and F. vesca.
a: F.×ananassa receptacle, b: F.×ananassa achenes, c: F. vesca receptacle, d: F. vesca achenes. red: DFR1, grey: DFR2. Data were calculated from three biological replicates and with error bars representing the standard deviation.