The use of mutants and transgenic plants to study nitrate assimilation


Michel Caboche, Laboratoire de Biologie Cellulaire, INRA, F-78026 Versailles Cedex, France.


The nitrate assimilatory pathway has been the matter of intensive genetic and molecular analysis over the past decade. Mutants impaired in the expression of nitrate reductase have been characterized in a number of plant species. Molecular analysis of the Nia gene coding for nitrate reductase has been the basis for a three-domain model of the structure of the enzyme, in agreement with biochemical and genetic data. Mutagenesis and antisense strategies have led to the description of nitrite reductase deficiencies. The molecular analysis of the corresponding Nii genes has provided invaluable information on the structure of nitrite reductase. Recently, a gene involved in nitrate uptake has also been identified. The regulation of the nitrate assimilatory pathway has been investigated. Analysis of the regulation of the pathway at the molecular level has shown evidence for the involvement of nitrate, light and/or sucrose, and reduced nitrogen in the regulation. Surprisingly, no bona fide regulatory mutant specific to this pathway has been identified so far in higher plants. This may reflect the redundancy of regulatory genes. The deregulated expression of one or the other step of the pathway obtained by ectopic expression of the corresponding genes is a new approach to study the physiological role of these regulations. Elements of the pathway have also been successfully used as transposon traps, or negatively selectable markers for other purposes. Finally, the identification at the molecular level of regulatory genes and structural elements involved in transport and storage of nitrate, or in the biosynthesis of cofactors of nitrate and nitrite reductases, will be the goal of the next decade.