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JXB Advance Access originally published online on May 11, 2006
Journal of Experimental Botany 2006 57(9):1957-1969; doi:10.1093/jxb/erj143
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© The Author [2006]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

RESEARCH PAPER

Metabolic engineering of rice with soybean isoflavone synthase for promoting nodulation gene expression in rhizobia

VS Sreevidya1,*, C Srinivasa Rao1,*, SB Sullia2, Jagdish K. Ladha1 and Pallavolu M. Reddy1,3,{dagger}

1International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines
2Department of Microbiology and Biotechnology, Bangalore University, Jnana Bharathi, Bangalore 560056, India
3Centro de Ciencias Genómicas, Programa de Genómica Funcional Eucariotes, Universidad Nacional Autónoma de México, Apdo. Postal 565-A, Cuernavaca, Morelos 62210, México

{dagger}To whom correspondence should be addressed in Mexico. E-mail: pmreddy{at}ccg.unam.mx

Isoflavonoids are derived from a flavonone intermediate, naringenin, that is ubiquitously present in plants, and play a critical role in plant development and defence response. Isoflavonoids secreted by the legumes also play an important role in promoting the formation of nitrogen-fixing nodules by symbiotic rhizobia. In these plants, the key enzyme that redirects phenylpropanoid pathway intermediates from flavonoids to isoflavonoids is the cytochrome P450 mono-oxygenase, isoflavone synthase. In an effort to develop a rice variety possessing the ability to induce nodulation (nod) genes in rhizobia, the IFS gene from soybean was incorporated into rice (Oryza sativa L. cv. Murasaki R86) under the control of the 35S promoter. The presence of IFS in transgenic rice was confirmed by PCR and Southern blot analysis. Analyses of the 35S-IFS transgenic lines demonstrated that the expression of the IFS gene led to the production of the isoflavone genistein in rice tissues. These results showed that the soybean IFS gene-expressed enzyme is active in the R86 rice plant, and that the naringenin intermediate of the anthocyanin pathway is available as a substrate for the introduced foreign enzyme. The genistein produced in rice cells was present in a glycoside form, indicating that endogenous glycosyltransferases were capable of recognizing genistein as a substrate. Studies with rhizobia demonstrated that the expression of isoflavone synthase confers rice plants with the ability to produce flavonoids that are able to induce nod gene expression, albeit to varied degrees, in different rhizobia.

Key words: Flavonoids, isoflavone synthase, nod gene induction, Oryza sativa, rhizobia


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