The biochemical basis of colour as an aesthetic quality in Citrus sinensis.
The development of fruit colour in the sweet orange (Citrus sinensis) is a cultivar characteristic affected by climate and environment. Although external colour is not always an indication of internal quality or maturity, it is probably the most important factor determining consumer acceptance. In the present investigation, efforts were made to determine the biochemical basis of colour as an aesthetic quality in C. sinensis 'Navel' and 'Valencia'. Furthermore, the changes in pigment content and composition during the period of colour development were recorded. Finally, in an attempt to manipulate citrus colour, the effect of dehydrating agents (alcohols), plant hormones, micro-nutrients and low temperature on flavedo carotenoid content was determined. Saponification of the two major colour-imparting components resolved by thin layer chromatography, followed by reversed-phase high performance liquid chromatography revealed that the principal colour-imparting pigments in C. sinensis flavedo are the yellow-coloured xanthophyll 9-Z-violaxanthin and the red C(30) apocarotenoid β-citraurin. Both pigments occur in the flavedo in esterified form. Identification of the chromophores was based on co-chromatography and online spectral analysis. The colour quality of mature fruit was dependant on the content and relative amounts of 9-Z-violaxanthin and Pcitraurin. Quantitative results revealed that increased colour intensity of citrus flavedo was associated with a decline in the 9-Z-violaxanthin : β-citraurin ratio from greater than 50 to below 10, and an increase in 9-Z-violaxanthin and β-citraurin content. Measurement of the mass and ratio of these pigments can be used to accurately colour-grade orange fruit for local and export markets. These parameters will also aid in the evaluation of colour manipulatory techniques. Visual colour break in C. sinensis appears to be associated with a minimum in total pigment as well as total carotenoid content. The period prior to colour break is characterised by a reduction in chlorophylls, carotenes and free xanthophylls usually associated with photosynthetic activity. Following colour break, a massive increase in xanthophyll acyl esters (particularly 9-Z-violaxanthin) is observed. Efforts were made to manipulate carotenoid content of citrus flavedo in vivo, with a view to manipulate fruit colour commercially. It was shown that the micro-nutrients tungsten and molybdenum, and the plant hormones abscisic acid and jasmonic acid increased carotenoid content of flavedo discs; whereas the plant hormone gibberellic acid decreased carotenoid levels. The dehydrating agents ethanol and butanol increased carotenoid content in whole fruit flavedo and flavedo discs. Optimum concentrations were shown to be 20-30% (v/v) for ethanol and 5-10% (v/v) for butanol.