Sulfation of nod factors via nodHPQ is nodD-independent in R.tropici CIAT899

Abstract

A cosmid from the Rhizobium tropici CIAT899 symbiotic plasmid, containing most of the nodulation genes described in this strain, has been isolated. Although this cosmid does not carry a nodD gene, it confers ability to heterologous Rhizobium spp. to nodulate R. tropici hosts (Phaseolus vulgaris, Macroptilium atropurpureum, and Leucaena leucocephala). The observed phenotype is due to constitutive expression of the nodABCSUIJ operon, which has lost its regulatory region and is expressed from a promoter present in the cloning vector. Thin-layer chromatography (TLC) analysis of the Nod factors produced by this construction shows that it is still capable of synthesizing sulfated compounds, suggesting that the nodHPQ genes are organized as an operon that is transcribed in a nodDindependent manner and is not regulated by flavonoids. Se ha aislado un cósmido del plásmido simbiótico de Rhizobium tropici CIAT899 que contiene la mayoría de los genes de nodulación descrito para esta estirpe, menos el gen regulador nodD. La introducción de este cósmido en una estirpe curada del plásmido simbiótico de R. tropici CIAT899 permite la nodulación en las plantas ensayadas (Phaseolus vulgaris, Macroptilium atropurpureum, y Leucaena leucocephala). El fenotipo observado se debe a la expresión constitutiva del operón nodABCSUIJ bajo el promotor del gen de resistencia a la kanamicina, que lleva el vector donde se ha clonado el fragmento de ADN. Análisis por cromatografia de capa fina demuestran que esta construcción es capaz de sulfatar el extremo reductor del factor Nod. Estas evidencias sugieren que los genes nodHPQ constituyen un operón, y que su expresión es independiente del gen regulador nodD.