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Fruits of 23 loquat ( Eriobotrya japonica Lindl.) cultivars, of which 11 were white-fleshed and 12 red-fleshed, were analyzed for color, carotenoid content, and vitamin A values. Color differences between two loquat groups were observed in the peel as well as in the flesh. beta-Carotene and lutein were the major carotenoids in the peel, which accounted for about 60% of the total colored carotenoids in both red- and white-fleshed cultivars. beta-Cryptoxanthin and, in some red-fleshed cultivars, beta-carotene were the most abundant carotenoids in the flesh, and in total, they accounted for over half of the colored carotenoids. Neoxanthin, violaxanthin, luteoxanthin, 9- cis-violaxanthin, phytoene, phytofluene, and zeta-carotene were also identified, while zeaxanthin, alpha-carotene, and lycopene were undetectable. Xanthophylls were highly esterified. On average, 1.3- and 10.8-fold higher levels of colored carotenoids were observed in the peel and flesh tissue of red-fleshed cultivars, respectively. The percentage of beta-carotene among colored carotenoids was higher in both the peel and the flesh of red-fleshed cultivars. Correlations between the levels of total colored carotenoids and the color indices were analyzed. The a* and the ratio of a*/ b* were positively correlated with the total content of colored carotenoids, while L*, b*, and H degrees correlated negatively. Vitamin A values, as retinol equivalents (RE), of loquat flesh were 0.49 and 8.77 microg/g DW (8.46 and 136.41 microg/100 g FW) on average for white- and red-fleshed cultivars, respectively. The RE values for the red-fleshed fruits were higher than fruits such as mango, red watermelon, papaya, and orange as reported in the literature, suggesting that loquat is an excellent source of provitamin A.

Polymethoxylated flavones (PMFs), which accumulate exclusively in fruit peel of citrus, play important physiological and pharmacological roles but the genetic basis for the methylation of flavonoids has not been fully elucidated in citrus. Here we characterize a caffeoyl-CoA O-methyltransferase-like enzyme, designated CrOMT1. The expression pattern of CrOMT1 was highly correlated with the concentration of the three major PMFs in two different citrus fruit tissues during fruit maturation. Exposure of fruit to UV-B radiation sharply increased the level of CrOMT1 transcripts and also led to the accumulation of three PMFs. The potential role of CrOMT1 was studied by testing the catalytic activity of recombinant CrOMT1 with numerous possible substrates in vitro. The enzyme could most efficiently methylate flavones with neighboring hydroxy moieties, with high catalytic efficiencies found with 6-OH- and 8-OH-containing compounds, preferences that correspond precisely with the essential methylation sites involved in the synthesis of the three naturally occurring PMFs in Citrus reticulata. This indicates that CrOMT1 is capable of in vitro methylation reactions required to synthesize PMFs in vivo. Furthermore, transient overexpression of CrOMT1 increased levels of the three major PMFs in fruit, indicating that CrOMT1 is likely to play an essential role in the biosynthesis of PMFs in citrus.

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Flavonoids are the most widespread polyphenolic compounds and are important dietary constituents present in horticultural crops such as fruits, vegetables, and tea. Natural flavonoids are responsible for important quality traits, such as food colors and beneficial dietary antioxidants and numerous investigations have shown that intake of flavonoids can reduce the incidence of various non-communicable diseases (NCDs). Analysis of the thousands of flavonoids reported so far has shown that different hydroxylation modifications affect their chemical properties and nutritional values. These diverse flavonoids can be classified based on different hydroxylation patterns in the B, C, A rings and multiple structure–activity analyses have shown that hydroxylation decoration at specific positions markedly enhances their bioactivities. This review focuses on current knowledge concerning hydroxylation of flavonoids catalyzed by several different types of hydroxylase enzymes. Flavonoid 3′-hydroxylase (F3’H) and flavonoid 3′5’-hydroxylase (F3’5’H) are important enzymes for the hydroxylation of the B ring of flavonoids. Flavanone 3-hydroxylase (F3H) is key for the hydroxylation of the C ring, while flavone 6-hydroxylase (F6H) and flavone 8-hydroxylase (F8H) are key enzymes for hydroxylation of the A ring. These key hydroxylases in the flavonoid biosynthesis pathway are promising targets for the future bioengineering of plants and mass production of flavonoids with designated hydroxylation patterns of high nutritional importance. In addition, hydroxylation in key places on the ring may help render flavonoids ready for degradation, the catabolic turnover of which may open the door for new lines of inquiry.

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