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A genomic approach to isoflavone biosynthesis in kudzu (Pueraria lobata)

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Abstract

Roots of kudzu (Pueraria lobata) are a rich source of isoflavone O- and C-glycosides. Although O-glycosylation of (iso)flavonoids has been well characterized at the molecular level, no plant isoflavonoid C-glycosyltransferase genes have yet been isolated. To address the biosynthesis of kudzu isoflavonoids, we generated 6,365 high-quality expressed sequence tags (ESTs) from a subtraction cDNA library constructed using RNA from roots that differentially accumulate puerarin. The ESTs were clustered into 722 TCs and 3,913 singletons, from which 15 family I glycosyltransferases (UGTs) were identified. Hierarchical clustering analysis of the expression patterns of these UGTs with isoflavone synthase (IFS) in a range of tissues identified UGTs with potential functions in isoflavone glycosylation. The open reading frames of these UGTs were expressed in E. coli for functional analysis, and one was shown to preferentially glycosylate isoflavones at the 7-O-position. In addition, ESTs corresponding to chalcone synthase, chalcone reductase, chalcone isomerase (CHI) and 2-hydroxyisoflavanone dehydratase were identified. Recombinant CHI proteins had high activities with both 6′-deoxy- and 6′-hydroxy chalcones, typical of Type II CHIs. Establishment of this EST database and identification of genes associated with kudzu isoflavone biosynthesis and glycosylation provide a new resource for metabolic engineering of bioactive kudzu isoflavones.

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Acknowledgments

We thank Drs. Ji He and Patrick Zhao for processing of ESTs, Dr. Marina Naoumkina for assistance in transcript cluster analysis, Tui Ray for assistance with qRT-PCR, Darla Boydston for assistance with artwork, and Drs. Yongzhen Pang and Rui Zhou for critical reading of the manuscript. This work was supported by the Oklahoma Center for the Advancement of Science and Technology (OCAST) Plant Science Program, and by the Samuel Roberts Noble Foundation.

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  1. Shujun Ge

    Present address: College of Agronomy, Agricultural University of Hebei, 2596 Lekai South Street, 071001, Baoding, Hebei, China

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  1. Plant Biology Division, Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK, 73401, USA

    XianZhi He, Jack W. Blount, Yuhong Tang & Richard A. Dixon

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He, X., Blount, J.W., Ge, S. et al. A genomic approach to isoflavone biosynthesis in kudzu (Pueraria lobata). Planta 233, 843–855 (2011). https://doi.org/10.1007/s00425-010-1344-1

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