Counteractive Action of Nitric Oxide on the Decrease of Nitrogenase Activity Induced by Enhanced Ultraviolet-B Radiation in Cyanobacterium

Abstract

The experimental enhancement of UV-B radiation resulted in damage to chlorophyll-a in Spirulina platensis 794, and the degree of this damage was modified by chemical treatments. The addition of 0.5 mM sodium nitroprusside (SNP), a donor of nitric oxide (NO), to cultures of Spirulina platensis 794 could markedly alleviate the damage to chlorophyll-a caused by enhanced ultraviolet-B radiation. Exposure of N(2)-fixing cyanobacterium Spirulina platensis 794 to enhanced ultraviolet-B radiation resulted in an intensity-dependent inhibition of nitrogenase activity. In cultured cells that were treated with 0.5 mM SNP and enhanced UV-B for 6 h, nitrogenase activity increased by 47.3% compared with UV-B treated control cells. SNP apparently counteracted the decrease in nitrogenase activity caused by UV-B stress. NAC (a free radical scavenger) significantly increased nitrogenase activity, but PTIO (a nitric oxide scavenger) decreased nitrogenase activity in UV-B treated S. platensis 794. Thus, the free radical scavenger NAC and NO may counteract the effects of enhanced UV-B radiation. The activity of UV-B-inhibited nitrogenase did not recover upon transfer of exposed cells to fluorescent light, suggesting that the inhibition may be due to specific inactivation of the enzyme. By experimentally manipulating the inhibitors of photosystem-II activity, it was demonstrated that nitrogenase activity in cyanobacterium S. platensis 794 is limited by the amount of reductant and ATP. This result further confirmed that nitrogenase activity requires a continued and abundant supply of suitable reductant and ATP for conversion of N(2) to NH(3). The effects of UV-B treatment on nitratase activity were also examined, and enhanced UV-B radiation increased nitratase activity. In addition, enhanced UV-B in combination with SNP and NAC resulted in significant increases in the activity of nitratase.