Use of NaCN to increase the growth of N₂-fixing bacteria in a model spermosphere mimicked by sucrose and amino acids leached by seeds

In order to study the establishment of a spermosphere, the C₂H₂ reduction activity of N₂-fixing bacteria isolated from river sand was examined in a simulated spermosphere in the river sand which contained sucrose, an amino acid mixture, and CN^- released from plant seeds. The sand incubated with 10^-10 to 10^-9 mol CN^- 30 g sand^-1 exhibited higher C₂H₂ reduction activity than that without CN^-. The change in the most probable number of N₂ fixers with increasing quantities of CN^- roughly corresponded to that in C₂H₂ reduction activity. However, the most probable number of non-N₂-fixing bacteria decreased except for CN^-- tolerant ones. Both C₂H₂ reduction activity and proliferation of the N₂ fixers isolated on a modified Burk's medium were almost similar to those in the bacteria in the sand. In contrast, the proliferation of some nonfixers decreased with an increasing CN^- concentration. C₂H₂ reduction activity of N₂ fixers cultured in combination with non-fixers exhibited a clear peak at 10^-7 M CN^- as for C₂H₂ reduction activity in the sand. We therefore speculate that cyanide evolved from seeds during a pregermination period may suppress the growth of general bacteria, but may promote the proliferation of N₂ fixers, thus contributing to the establishment of a spermosphere.