A procedure for in vitro nodulation studies

Abstract

Standardised in vitro conditions yielding successful, synchronised, reproducible cycles of symbiotic nitrogen fixation are essential experimental tools for investigating the different stages of the Rhizobium-legume interaction. A standard in vitro procedure can facilitate analysis of the kinetics of phenotype at the molecular level occurring after Rhizobium infection. This is essential when comparing, for instance, the expression pattern of nodulin or marker genes or the phenotypes of wild type and mutant lines. We present here a feasible, reliable procedure for in vitro nodulation studies of L. japonicus. All the different steps of the symbiotic interaction have been analysed and different parameters of plant growth followed to demonstrate the reproducibility of the procedure.