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First published online January 23, 2003; 10.1104/pp.004820

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Plant Physiol, March 2003, Vol. 131, pp. 941-951

A Cluster of Genes Encodes the Two Types of Chalcone Isomerase Involved in the Biosynthesis of General Flavonoids and Legume-Specific 5-Deoxy(iso)flavonoids in Lotus japonicus1

Norimoto Shimada, Toshio Aoki, Shusei Sato, Yasukazu Nakamura, Satoshi Tabata, and Shin-ichi Ayabe*

Department of Applied Biological Sciences, Nihon University, Fujisawa, Kanagawa 252-8510, Japan (N.S., T.A., S.A.); and Kazusa DNA Research Institute, 1532-3 Yana, Kisarazu, Chiba 292-0812, Japan (S.S., Y.N., S.T.)

Leguminous plants produce 5-deoxyflavonoids and 5-deoxyisoflavonoids that play essential roles in legume-microbe interactions. Together with chalcone polyketide reductase and cytochrome P450 2-hydroxyisoflavanone synthase, the chalcone isomerase (CHI) of leguminous plants is fundamental in the construction of these ecophysiologically active flavonoids. Although CHIs of nonleguminous plants isomerize only 6'-hydroxychalcone to 5-hydroxyflavanone (CHIs with this function are referred to as type I), leguminous CHIs convert both 6'-deoxychalcone and 6'-hydroxychalcone to 5-deoxyflavanone and 5-hydroxyflavanone, respectively (referred to as type II). In this study, we isolated multiple CHI cDNAs (cCHI1-cCHI3) from a model legume, Lotus japonicus. In contrast to previous observations, the amino acid sequence of CHI2 was highly homologous to nonleguminous CHIs, whereas CHI1 and CHI3 were the conventional leguminous type. Furthermore, genome sequence analysis revealed that four CHI genes (CHI1-3 and a putative gene, CHI4) form a tandem cluster within 15 kb. Biochemical analysis with recombinant CHIs expressed in Escherichia coli confirmed that CHI1 and CHI3 are type II CHIs and that CHI2 is a type I CHI. The occurrence of both types of CHIs is probably common in leguminous plants, and it was suggested that type II CHIs evolved from an ancestral CHI by gene duplication and began to produce 5-deoxy(iso)flavonoids along with the establishment of the Fabaceae.


1 This work was supported by the Ministry of Education, Sports, Science and Culture of Japan (Grant-in-Aid for Scientific Research on Priority Area [A] no. 12045261).

* Corresponding author; e-mail ayabe{at}brs.nihon-u.ac.jp; fax 81-466-80-1141.

© 2003 American Society of Plant Biologists



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