The negative effect of nitrate on gametogenesis is independent of nitrate assimilation in Chlamydomonas reinhardtii

Abstract.

 The effect of nitrate on gamete differentiation as well as on the expression of genes involved in gametogenesis, nitrogen scavenging, and nitrate assimilation has been analyzed in wild-type and mutant strains of Chlamydomonas reinhardtii. Nitrate prevented gamete formation from wild-type strains and caused a strong reduction in the number of zygotes recovered in genetic crosses between nitrate-assimilation-deficient mutants, thus suggesting that nitrate by itself is providing a negative regulatory signal for the sexual differentiation of the alga. Addition of nitrate at low concentrations to wild-type cells, after an initial period of nitrogen starvation, resulted in a drastic decrease in transcript levels of both nitrate-assimilation genes (NIA1 and NRT2;1) and genes induced after N-starvation (NCG2 and NCG4). This strong effect of nitrate was due to its assimilation products since it was not evident in nitrate-assimilation mutants. A slight negative effect of nitrate on NCG4 expression was only observed in the mutant. Nitrate by itself was also found to provide a negative signal for the expression of gamete-specific genes (GAS3 and GAS18) in mutants incapable of assimilating nitrate.