The nitrate reductase (NR) gene
niaA of the oomycete
Phytophthora infestans was selected from a gene library by heterologous hybridization.
NiaA occurs as a single-copy gene and its expression is regulated by the nitrogen source. The nucleotide sequence of
niaA was determined and comparison of the deduced amino-acid sequence of 902 residues with NRs of higher fungi and plants revealed a significant homology, particularly within the three cofactor-binding domains for molybdenum, heme and FAD. The
P. infestans niaA gene was used as a model gene to test whether oomycete genes are functional in the ascomycete
Aspergillus nidulans, a fungus which is highly accessible for molecular genetic studies. The complete
niaA gene was stably integrated into the genome of a
nia
- deletion mutant of
A. nidulans. However, transformants containing one or more copies of the
niaA gene were not able to complement the
nia
- mutant. This suggests that there is no functional expression of the introduced
niaA gene in
A. nidulans. In addition, the activity of two other oomycete gene promoters was analyzed in a transient expression assay. Plasmids containing chimaeric genes with the promoter of the
P. infestans ubiquitin gene
ubi3R, or the
Bremia lactucae ham34 gene, fused to the coding sequence of the
Escherichia coli 
-glucuronidase (GUS) reporter gene, were transferred to
A. nidulans protoplasts. No significant GUS activity was detectable indicating that the
ubi3R and
ham34 promoters are not active in
A. nidulans. Apparently, the regulatory sequences which are sufficient for gene activation in oomycetes are not functional in the ascomycete
A. nidulans.
Key words
Phytophthora infestans
- Nitrate reductase gene - Transformation - Oomycete
Communicated by P. J. G. M. de Wit