Expression of the legume symbiotic lectin genes psl and gs52 promotes rhizobial colonization of roots in rice

Transgenic rice (Oryza sativa L. cv. Murasaki) carrying genes encoding pea (Pisum sativum) lectin (PSL) or wild-soybean (Glycine soja) lectin-nucleotide phosphohydrolase (GS52) were inoculated with Rhizobium leguminosarum bv. viciae or Bradyrhizobium japonicum USDA110, respectively, as well as with Rhizobium sp. NGR234, and root colonization was assessed in comparison to comparably inoculated control plants. The data showed that expression of PSL and GS52 significantly promoted rhizobial colonization of root epidermal cells including root hairs in rice. In addition, in the case of R. leguminosarum bv. viciae and B. japonicum USDA110 colonization of the psl and gs52 transgenic rice plants, respectively, the bacterial cells were found to preferentially home towards and aggregate maximally at the root hair tip regions rather than on the root hair "stalks". The above data suggest that the lectins PSL and GS52, which participate in rhizobial recognition by root epidermal cells in pea and soybean, respectively, are also able to facilitate rhizobial attachment and colonization of the epidermal cells in rice roots. Moreover, aggregation of R. leguminosarum bv. viciae and B. japonicum USDA110 cells preferentially at root hair tip regions suggest that similar to legumes, the PSL and GS52 lectins are targeted to the root hair tips in transgenic rice, enabling higher bacterial attachment/colonization at the tip region. Rhizobial colonization at root hair tips in the psl and gs52 rice plants frequently led to the localized dissolution of the cell wall creating perforations at the tip region. It is likely that the presence of lectins, such as PSL and GS52 leads to structural modifications in cell wall organization of the root hair/epidermal cells, making them prone to localized dissolution by the hydrolytic activity of compatible rhizobia to permit invasion of the root cells. © 2005 Elsevier Ireland Ltd. All rights reserved

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

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. © The Author 2006. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved