Chalcone Synthase Transcripts Are Detected in Alfalfa Root Hairs Following Inoculation with Wild-Type Rhizobium meliloti

Flavonoids are involved in a number of critical events in the interaction between nitrogen-fixing bacteria and legumes. To get a better understanding of the importance of flavonoids in the earliest stages of the alfalfa-Rhizobium meliloti symbiosis, we followed the expression of two chal-cone synthase (CHS) gene family members as well as of chalcone isomerase (CHI) and isoflavone reductase (IFR) genes. CHS transcripts increased 2 to 4 dpi (days post-inoculation) with wild-type rhizobia, but not after inoculation with the heterologous R. leguminosarum bv. trifolii or with an exopolysaccharide (exo) mutant of R. meliloti. CHS transcripts were detected in the root hairs and epidermal cells of the root hair zone, and infrequently in nodule pri-mordia. Insignificant CHI and IFR mRNA accumulation over control levels was observed in response to rhizobial inoculation. The slight increase in CHS transcript accumulation following wild-type R. meliloti inoculation was correlated with an observed increase in root flavonoid content as well as a change in the nod gene-inducing activity of the root exudate. The nod gene-inducing flavonoids exuded from wild-type rhizobia-inoculated roots were identified as 4′, 7-dihydroxyflavone and 4, 4′ dihydroxy-2′-methoxychalcone. Although there was a slight increase over the uninoculated controls in the level of medicarpin-3-O-glucoside 6″-O-malonate (MGM) in extracts of roots inoculated with rhizobia, IFR transcript accumulation was not significantly elevated over that of the controls. Moreover, no medicarpin aglycone was detected in the inoculated roots. Thus, although inoculation with wild-type rhizobia triggers some of the genes induced during an interaction between a host and a pathogen, the expression of these genes in the Rhizobium-legume interaction is at a very low level, suggesting that rhizobia have evolved a mechanism(s) to avoid triggering the host's defense responses.