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. 2021 Jul 15;38(11):4659–4673. doi: 10.1093/molbev/msab213

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© The Author(s) 2021. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Fig. 6. — IaCYP708A15v2 catalyzes 22-hydroxylation in the cucurbitacin biosynthesis. (A) Hierarchical cluster heat map of expression patterns of the 287 P450 contigs and five oxidosqualene cyclases expressed in the roots, petals, and stems of Iberis amara. Putative biosynthetic P450 genes involved in cucurbitacin biosynthesis are marked by a yellow box. (B) Doughnut chart was drawn based on the exact number of genes from each P450 subfamily of I. amara (outer doughnut) and Arabidopsis thaliana (inner doughnut). Each subfamily was color coded. In particular, the CYP72A, CYP708A, CYP81D, and CYP707A subfamilies were expanded in I. amara as compared with A. thaliana. (C) GC-MS chromatograms of extracts from Nicotiana benthamiana leaves agro-infiltrated with EV control and vectors expressing IaCPQ, IaCPQ+CYP708A16, IaCPQ+CYP708A15v2, IaCPQ+CYP708A16+CYP708A15v2, and CYP708A16+CYP708A15v2. Cucurbitadienol and 16β-hydroxy-cucurbitadienol are indicated with black arrows. The new peak that appeared only in the extract from IaCPQ+CYP708A16+CYP708A15v2 expressing leaves is indicated by a red arrow.