mep.claudie Dhuique:mep.DB 10/09/09 16:57 Page 1 Effect of genotype and geographical origins on carotenoid content from Citrus sweet orange juices

C. Dhuique-Mayer1*, A.L Fanciullino2, ITRUS juices, such as orange juices, represent C. Dubois3, P. Ollitrault4 significant sources of carotenoids that could contribute, to the 1CIRAD-Dept persyst-UMR Qualisud, TA B95/16, 2 INRA UR Geqa, site de San Guliano, beneficial health effects of this fruit product. Genetic and F-20230, San Guliano, France 3,4 Dept BIOS UPR 75, 34398 Montpellier cedex 5, environmental factors influenced the carotenoid France ([email protected]). composition of citrus juices. In order to assess the * corresponding author [email protected] effect of genotype and geographical origins, on carotenoid content from citrus juices, a selection of three orange varieties cultivated in Mediterranean, subtropical and tropical areas were analysed.

Material and Method Juices from three sweet orange cultivars, i.e. Pera, Sanguinelli and Valencia (Citrus sinensis (L.) Osbeck) belonging to the same Citrus accessions and cultivated in Mediterranean (Corsica), and tropical areas (New Caledonia, Tahiti, Brazil, Cuba and Costa-Rica). All juices were analysed by HPLC using a C30 column according to a previous method (1). Representative samples were collected from different countries at commercial maturity using maturity index as indicator. Statistical analysis was used to evaluate significance of variety and year effect and to classify juices (SAS 9.1 Windows USA, @DARwin 5.0 cirad and XLSTAT 7.0 France).

Results and discussion Table 1. Carotenoid contents (mg/L) and vitamin A values in three varieties of Corsican orange juices, mean over 3 years. Orange varieties phytoene phytofluene β−carotene β−cryptoxanthin zeaxanthin Cis-violaxanthin lutein Vit A* TT Annual carotenoid content variations in Corsican fruits were found to be Valencia CO 0.48 0.79 0.44 2.1 1.77 5.02 1.37 124 0.10 0.20 0.10 0.40 0.20 0.40 0.40 limited compared to variations due to varietal influences (p < 0.05). The genetic Sanguin CO 0.94 1.01 0.43 4.1 1.86 6.37 1.79 207 variability among sweet orange cultivars was associated with significant 0.07 0.05 0.10 0.10 0.16 1.20 0.20 Pera CO 0.72 0.67 0.5 3.17 2.37 5.26 1.5 174 variations in the contents of four carotenoids (cis-violaxanthin, -cryptoxanthin, 0.20 0.10 0.10 0.40 0.60 0.70 0.40 phytoene and phytofluene). Sanguinelli and Pera cultivars revealedβ significantly * Expressed as retinol activity equivalent L-1 (Conversion factor 1 µg RAE = 12 µg -carotene+24 µg -cryptoxanthine) higher provitamin A carotenoid than Valencia variety (table 1). TT Variations in Valencia oranges from five geographical origins were illustrated by a dissimilarity tree (figure 1) which was drawn up with the content of seven carotenoids, listed in table 1, underlined that the quantitative compositions were more geographical origin-dependent than annual variation- V.CostaR dependent. V.NCal V.Tahiti TT Influence of environmental conditions on carotenoid contents was particularly high for sweet V.Cors3 oranges. Principal component analysis (PCA) showed that Mediterranean sweet oranges, with high carotenoid contents, are clearly differentiated from those of all other origins (figure 2). Strong V.CubaV.Cuba V.Cors2 V.Cors1 correlations were observed between -cryptoxanthin, phytofluene and phytoene (r = 0.931) and 0 2. between -carotene and phytoene (0.918).β To complete this PCA, the graph from the figure 3 Corsica V.NCal: Valencia New Caledonia; V.CostaR: Valencia Costa Rica. highlightedβ that carotenoid profiles varied as a function of the two principal carotenoids ( -cryptoxanthin and cis-violaxanthin) with respect to origins (New-Caledonia, Corsica). Figure 1. Classification of Valencia β orange juices from five different origins. The tree was constructed according to the Neighbour-joining method using Factorial map of samples (F1 x F2 plane : 88 %) Correlation circle Euclidean distances. 1

1,5 0,8 lutein San.NC 0,6 1 Val.C5 Per.C5 beta- Val.CR Per.BR San.C5 0,4 Val.NC cryptoxanthin 0,5 0,2 zeaxanthin 6 violaxanthin 0 0 β−carotene phytofluene Val.TA Val.C3 San.C3 -0,2 4 β−cryptoxanthin phytofluene violaxanthin -- axis F2-0,5 (9 %) --> -- axis F2 (9 %)-0,4 --> phytoene Val.C4 San.C4 -0,6 2 -1 Val.CU Per.C3 -0,8

-1,5 -1 0 -4 -3 -2 -1 0 1 2 3 4 -1 -0,5 0 0,5 1 -- axe F1 (79 %) --> -- axe F1 (79 %) --> Corsica phytoene zeaxanthin Tropical zones

Figure 2. Orange juice origin differentiation according to PCA based on seven carotenoids as variables. Corsica Per: Pera oranges; San: Sanguinelli oranges; Val: Valencia oranges. BR: Brazil; C3: Corsica year 1; New Caledonia C4: Corsica year 2; C5: Corsica year 3; CR: Costa Rica; CU: Cuba; NC: New Caledonia; TA: Tahiti. beta-carotene lutein Martine, Duportal September 2009 Figure 3. “Radar” chart showing variations in carotenoid profiles of orange with respect to origins Conclusion (New Caledonia, Corsica 2005). UR results underlined the importance of environmental factors in carotenoid synthesis and accumulation in citrus fruits. Interactions between Citrus accessions and environmental

1. DHUIQUE-MAYER C., CARIS-VEYRAT C., Design and production: CIRAD, OLLITRAULT P., CURK F. , AMIOT M.J. conditions may occur with respect to carotenoid biosynthesis in juice sacs. We suggest that Centre (2005) Varietal and interspecific O de coopération influence on micronutrient contents in environmental conditions play a major role on the concentration of secondary metabolites of internationale citrus from the Mediterranean area. nutritional interest, such as -cryptoxanthin, the main provitamin A carotenoid in orange citrus juices. en recherche Journal of Agricultural and Food agronomique pour le Chemistry, 53, 2140-2145. β développement R e f e r e n c e