c 1997 Kluwer Academic Publishers. Printed in the Netherlands. Biological nitrogen fixation in non-leguminous field crops: Facilitating the evolution of an effective association between Azospirillum and wheat

Recent advances towards achieving significant nitrogen fixation by diazotrophs in symbioses with cereals are reviewed, referring to the literature on the evolution of effective symbioses involving rhizobia and Frankia as microsymbionts. Data indicating that strains of Acetobacter and Herbaspirillum colonizing specific cultivars of sugarcane as endophytes make a significant contribution to the nitrogen economy of this crop improves the prospects that similar associative systems may be developed for other gramineous species such as rice and wheat. By contrast, the transfer of nodulation genes similar to those in legumes or Parasponia to achieve nodulation in crops like rice and wheat is considered to be a more ambitious and distant goal. Progress in developing an effective associative system for cereals has been materially assisted by the development of genetic tools based on the application of lacZ and gusA fusions with the promoters of genes associated with nitrogen fixation. These reporter genes have provided clear evidence that ‘crack-entry ’ at the points of emergence of lateral roots or of 2,4-D induced para-nodules is the most significant route of endophytic colonization. Furthermore, using the laboratory model of para-nodulated wheat, there is now evidence that the ability of azospirilla and other nitrogen fixing bacteria to colonize extensively as endophytes can be genetically controlled. The most successful strain of Azospirillum brasilense (Sp7-S) for endophytic colonization and nitrogen fixation in wheat seedlings is a mutant with reduce