Physiological and morphological aspects of interactions between Rhizobium meliloti and alfalfa (Medicago sativa) in association with Azospirillum brasilense

In a 50-L pot experiment with Medicago sativa grown under nonsterile conditions, a combined treatment of Azospirillum and Rhizobium was measured against soil inoculated with Rhizobium or Azospirillum alone or a control with a low background level of autochthonous rhizobia. The combined treatment significantly increased the shoot length and weight at 6 weeks and the regrowth shoot weight at 14 weeks when compared with the treatment with Rhizobium alone. In 1.5-L pots in which gnotobiotic conditions were maintained, the combined treatment led to more nodules on the main root at intermediate Rhizobium concentrations, and a greater root surface area at intermediate and high Rhizobium concentrations after 2 weeks but not after 4 weeks. In pouch-grown seedlings, plants were inoculated with either Rhizobium alone or in combination with Azospirillum or applied together with a flavonoid, luteolin (a nodulation gene inducer), or with a cytokinin, benzyl adenine. Luteolin had similar effects to those of Azospirillum in increasing the main root nodule number and the total nodule number. With Fahraeus slides, a significant increase was observed in the number of root hairs and the root diameter in the presence of Azospirillum as compared with the control and Rhizobium alone. There was no increase in the total number of infection threads; however, the combined treatment caused a significant decrease in the percentage of infected root hairs.