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Sweetpotato  Ipomoea batatas  is known as a hexaploid species. Here, we analyzed carotenoids contained in the leaves and tubers of sweetpotato cultivars ‘White Star’ (WS) and W71. These cultivars were found to contain several carotenoids unique to sweetpotato tubers such as β-carotene-5,6,5′,8′-diepoxide and β-carotene-5,8-epoxide. Next, we isolated two kinds of carotene cyclase genes that encode lycopene β- and ε-cyclases from the WS and W71 leaves, by RT-PCR and subsequent RACE. Two and three lycopene β-cyclase gene sequences were, respectively, isolated from WS, named  IbLCYb1 ,  2 , and from W71,  IbLCYb3 ,  4 ,  5 . Meanwhile, only a single lycopene ε-cyclase gene sequence, designated  IbLCYe , was isolated from both WS and W71. These genes were separately introduced into a lycopene-synthesizing  Escherichia coli  transformed with the  Pantoea ananatis crtE ,  crtB  and  crtI  genes, followed by HPLC analysis. β-Carotene was detected in  E. coli  cells that carried  IbLCYb1 - 4 , indicating that the  IbLCYb1 - 4  genes encode lycopene β-cyclase. Meanwhile, the introduction of  IbLCYe  into the lycopene-synthesizing  E. coli  led to efficient production of δ-carotene with a monocyclic ε-ring, providing evidence that the  IbLCYe  gene codes for lycopene ε-(mono)cyclase. Expression of the β- and ε-cyclase genes was analyzed as well.

Carotenoid analysis of sweetpotato Ipomoea batatas and functional identification of its lycopene β- and ε-cyclase genes