Center for Environmental and Estuarine Studies, University of Maryland, Chesapeake Biological Laboratory, Solomons, Maryland 20688-0038; New England BioLabs, Inc., Beverly, Massachusetts 01915 ; and Marine Sciences Research Center, State University of New York, Stony Brook, New York 11794-5000.
Appl Environ Microbiol. 1990 Nov;56(11):3532-6. doi: 10.1128/aem.56.11.3532-3536.1990.
Natural populations of the nonheterocystous marine cyanobacterium Trichodesmium thiebautii exhibit a diel periodicity in nitrogenase activity (NA). NA "turns on" near dawn and "turns off" near dusk, independent of photic conditions. Chloramphenicol (CAP) and ammonium prevented turn on of NA in T. thiebautii when added to samples collected before dawn but were progressively less effective in inhibiting NA in samples collected later in the morning. In samples collected after turn on, activities declined with time with both CAP and ammonium treatments, with ammonium having a stronger effect. In contrast, CAP added to samples collected in late afternoon prolonged NA, compared with controls, which turned off. Direct analysis of the presence of the Fe protein of nitrogenase in T. thiebautii by using a Western immunoblot procedure found a strong protein band present in samples collected after 0800 h through the late evening but little or no Fe protein in samples collected within the 2 to 4 h preceding dawn. We conclude that the diel cycle of NA in T. thiebautii results from de novo synthesis of nitrogenase each morning and from the inactivation and degradation of nitrogenase in the late afternoon and night.
非异形固氮海洋蓝藻束毛藻的自然种群表现出固氮酶活性(NA)的昼夜周期性。NA 在黎明附近“开启”,在黄昏附近“关闭”,与光照条件无关。当在黎明前收集的样品中添加氯霉素(CAP)和铵时,CAP 和铵可防止束毛藻中 NA 的“开启”,但在上午晚些时候收集的样品中抑制 NA 的效果逐渐减弱。在“开启”后收集的样品中,CAP 和铵处理均随时间推移而使活性下降,而铵的作用更强。相比之下,与对照相比,下午晚些时候添加到样品中的 CAP 延长了 NA 的时间,而对照则关闭了。通过使用 Western 免疫印迹程序直接分析束毛藻中固氮酶的 Fe 蛋白的存在情况,发现从 0800 小时到深夜,在收集的样品中存在强蛋白带,但在黎明前 2 至 4 小时内收集的样品中几乎没有或没有 Fe 蛋白。我们得出结论,束毛藻中 NA 的昼夜周期是由于每天早晨固氮酶的从头合成以及下午和晚上固氮酶的失活和降解所致。