Suppression of an elicitor-induced oxidative burst reaction in Medicago sativa cell cultures by Sinorhizobium meliloti lipopolysaccharides

Albus U, Baier R, Holst O, Pühler A, Niehaus K. Suppression of an elicitor-induced oxidative burst reaction in Medicago sativa cell cultures by Sinorhizobium meliloti lipopolysaccharides. New Phytologist . 2001;151(3):597-606.The biological activity of lipopolysaccharides (LPS) from the symbiotic soil bacterium Sinorhizobium meliloti was analysed in cell cultures of the host plant Medicago sativa (alfalfa) and the nonhost plant Nicotiana tabacum (tobacco). LPS of S. meliloti were purified and chemically characterized. Alfalfa and tobacco suspension cell cultures responded to yeast elicitors with an alkalinization of the culture medium and the induction of an oxidative burst. This assay was used to study the biological activity of isolated LPS. In alfalfa cell cultures the simultaneous addition of purified LPS of S. meliloti suppressed the elicitor induced alkalinization and oxidative burst reaction. Cell cultures of the nonhost tobacco reacted differently to the application of S. meliloti LIPS. In these cell cultures, the S. meliloti LIPS itself caused an alkalinization of the culture medium and an oxidative burst reaction. S. meliloti LPS released from the bacterial surface might function as a specific signal molecule, promoting the symbiotic interaction and suppressing a pathogenic response in the host plant, alfalfa. (C) New Phytologist (2001)

Genetic Characterization of a Sinorhizobium meliloti Chromosomal Region Involved in Lipopolysaccharide Biosynthesis

The genetic characterization of a 5.5-kb chromosomal region of Sinorhizobium meliloti 2011 that contains lpsB, a gene required for the normal development of symbiosis with Medicago spp., is presented. The nucleotide sequence of this DNA fragment revealed the presence of six genes: greA and lpsB, transcribed in the forward direction; and lpsE, lpsD, lpsC, and lrp, transcribed in the reverse direction. Except for lpsB, none of the lps genes were relevant for nodulation and nitrogen fixation. Analysis of the transcriptional organization of lpsB showed that greA and lpsB are part of separate transcriptional units, which is in agreement with the finding of a DNA stretch homologous to a “nonnitrogen” promoter consensus sequence between greA and lpsB. The opposite orientation of lpsB with respect to its first downstream coding sequence, lpsE, indicated that the altered LPS and the defective symbiosis of lpsB mutants are both consequences of a primary nonpolar defect in a single gene. Global sequence comparisons revealed that the greA-lpsB and lrp genes of S. meliloti have a genetic organization similar to that of their homologous loci in R. leguminosarum bv. viciae. In particular, high sequence similarity was found between the translation product of lpsB and a core-related biosynthetic mannosyltransferase of R. leguminosarum bv. viciae encoded by the lpcC gene. The functional relationship between these two genes was demonstrated in genetic complementation experiments in which the S. meliloti lpsB gene restored the wild-type LPS phenotype when introduced into lpcC mutants of R. leguminosarum. These results support the view that S. meliloti lpsB also encodes a mannosyltransferase that participates in the biosynthesis of the LPS core. Evidence is provided for the presence of other lpsB-homologous sequences in several members of the family Rhizobiaceae