Marco E, Ohad N, Schwarz R, Lieman-Hurwitz J, Gabay C, Kaplan A
Department of Botany, Hebrew University of Jerusalem, Israel.
Plant Physiol. 1993 Mar;101(3):1047-53. doi: 10.1104/pp.101.3.1047.
The high-concentration CO2-requiring mutant N5 of Synechococcus sp. PCC 7942 was obtained by the insertion of a kanamycin-resistant gene at the EcoRI site, 12.4 kb upstream of rbc. The mutant is unable to accumulate inorganic carbon internally and exhibits very low apparent photosynthetic affinity for inorganic carbon but a photosynthetic Vmax similar to that of the wild type. Sequence and northern analyses showed that the insertion inactivated a gene highly homologous to ndhB, encoding subunit II of NADH dehydrogenase in Synechocystis sp. PCC 6803 (T. Ogawa [1991] Proc Natl Acad Sci USA 88: 4275-4279). When the mutant and the wild-type cells were exposed to 5% CO2 in air, their photosynthetic electron transfer capabilities, as revealed by fluorescence and thermoluminescence measurements, were similar. On the other hand, a significant decrease in variable fluorescence was observed when the mutant (but not the wild-type) cells were exposed to low CO2 under continuous light. The same treatment also resulted in a shift (from 38-27 degrees C) in the temperature at which the maximal thermoluminescence emission signal was obtained in the mutant but not in the wild type. These results may indicate that subunit II of NADH dehydrogenase is essential for the functional operation of the photosynthetic electron transport in Synechococcus under low but not high levels of CO2. We suggest that the inability to accumulate inorganic carbon under air conditions stems from disrupture of electron transport in this mutant.
通过在rbc上游12.4 kb处的EcoRI位点插入卡那霉素抗性基因,获得了集胞藻属PCC 7942的高浓度二氧化碳需求突变体N5。该突变体无法在细胞内积累无机碳,对无机碳表现出非常低的表观光合亲和力,但光合最大反应速率与野生型相似。序列分析和Northern分析表明,该插入使一个与ndhB高度同源的基因失活,ndhB编码集胞藻属PCC 6803中NADH脱氢酶的亚基II(T. 小川[1991]《美国国家科学院院刊》88: 4275 - 4279)。当突变体和野生型细胞暴露于空气中5%的二氧化碳时,荧光和热发光测量显示它们的光合电子传递能力相似。另一方面,当突变体(而非野生型)细胞在连续光照下暴露于低二氧化碳环境时,观察到可变荧光显著降低。相同处理还导致突变体获得最大热发光发射信号的温度发生了偏移(从38℃变为27℃),而野生型则没有。这些结果可能表明,在低水平而非高水平二氧化碳条件下,NADH脱氢酶的亚基II对于集胞藻光合电子传递的功能运行至关重要。我们认为,该突变体在空气条件下无法积累无机碳是由于电子传递受到破坏所致。