Elhai J, Wolk C P
MSU-DOE Plant Research Laboratory, Michigan State University, East Lansing 48824-1312.
EMBO J. 1990 Oct;9(10):3379-88. doi: 10.1002/j.1460-2075.1990.tb07539.x.
Depriving the cyanobacterium Anabaena of fixed nitrogen induces the differentiation of heterocysts at intervals along its filaments. To test whether the oxygen-deficient conditions believed to prevail within mature heterocysts are sufficient, in the absence of fixed nitrogen, to elicit the expression of nitrogenase, PnifHDK was fused transcriptionally to luxAB (encoding luciferase). Expression, monitored from individual cells as light emission, was localized (with a resolution of approximately 1 micron) to differentiated cells, whether or not oxygen was present. Anabaena PCC 7118 is a heterocystless mutant strain that is known to fix nitrogen when deprived of combined nitrogen under anaerobic conditions. Three lines of evidence indicate that the mutant has retained the ability to develop a pattern despite its inability to make heterocysts. First, morphologically distinct cells appear at nonrandom intervals when filaments are starved of nitrogen. Second, these cells, like heterocysts, have little or no phycocyanin-dependent fluorescence. Third, nitrogen-starved filaments fragment, with fragment lengths similar to the spacing normally seen between heterocysts. Expression of PnifHDK-luxAB was largely confined to differentiated cells in the mutant as in the wild-type strain. These results provide evidence for a causal relationship between development and transcriptional events in Anabaena.
剥夺蓝藻鱼腥藻的固定氮会诱导其藻丝上间隔出现异形胞分化。为了测试在缺乏固定氮的情况下,成熟异形胞内被认为普遍存在的缺氧条件是否足以引发固氮酶的表达,将PnifHDK转录融合到luxAB(编码荧光素酶)上。通过监测单个细胞的发光来检测表达情况,结果表明,无论是否存在氧气,表达都定位在分化细胞中(分辨率约为1微米)。鱼腥藻PCC 7118是一种无异形胞的突变菌株,已知在厌氧条件下剥夺化合态氮时能固定氮。三条证据表明,尽管该突变体无法形成异形胞,但仍保留了发育模式的能力。第一,当藻丝缺乏氮时,形态上不同的细胞会以非随机的间隔出现。第二,这些细胞与异形胞一样,几乎没有或没有藻蓝蛋白依赖性荧光。第三,缺氮的藻丝会断裂,片段长度与正常情况下异形胞之间的间距相似。与野生型菌株一样,PnifHDK-luxAB在突变体中的表达主要局限于分化细胞。这些结果为鱼腥藻发育与转录事件之间的因果关系提供了证据。