Tichý Martin, Bečková Martina, Kopečná Jana, Noda Judith, Sobotka Roman, Komenda Josef
Laboratory of Photosynthesis, Institute of Microbiology, Academy of Sciences of the Czech Republic, Center AlgatechTřeboň, Czech Republic; Faculty of Science, University of South BohemiaČeské Budějovice, Czech Republic.
Laboratory of Photosynthesis, Institute of Microbiology, Academy of Sciences of the Czech Republic, Center Algatech Třeboň, Czech Republic.
Front Plant Sci. 2016 May 12;7:648. doi: 10.3389/fpls.2016.00648. eCollection 2016.
Cyanobacterium Synechocystis PCC 6803 represents a favored model organism for photosynthetic studies. Its easy transformability allowed construction of a vast number of Synechocystis mutants including many photosynthetically incompetent ones. However, it became clear that there is already a spectrum of Synechocystis "wild-type" substrains with apparently different phenotypes. Here, we analyzed organization of photosynthetic membrane complexes in a standard motile Pasteur collection strain termed PCC and two non-motile glucose-tolerant substrains (named here GT-P and GT-W) previously used as genetic backgrounds for construction of many photosynthetic site directed mutants. Although, both the GT-P and GT-W strains were derived from the same strain constructed and described by Williams in 1988, only GT-P was similar in pigmentation and in the compositions of Photosystem II (PSII) and Photosystem I (PSI) complexes to PCC. In contrast, GT-W contained much more carotenoids but significantly less chlorophyll (Chl), which was reflected by lower level of dimeric PSII and especially trimeric PSI. We found that GT-W was deficient in Chl biosynthesis and contained unusually high level of unassembled D1-D2 reaction center, CP47 and especially CP43. Another specific feature of GT-W was a several fold increase in the level of the Ycf39-Hlip complex previously postulated to participate in the recycling of Chl molecules. Genome re-sequencing revealed that the phenotype of GT-W is related to the tandem duplication of a large region of the chromosome that contains 100 genes including ones encoding D1, Psb28, and other PSII-related proteins as well as Mg-protoporphyrin methylester cyclase (Cycl). Interestingly, the duplication was completely eliminated after keeping GT-W cells on agar plates under photoautotrophic conditions for several months. The GT-W strain without a duplication showed no obvious defects in PSII assembly and resembled the GT-P substrain. Although, we do not exactly know how the duplication affected the GT-W phenotype, we hypothesize that changed stoichiometry of protein components of PSII and Chl biosynthetic machinery encoded by the duplicated region impaired proper assembly and functioning of these multi-subunit complexes. The study also emphasizes the crucial importance of a proper control strain for evaluating Synechocystis mutants.
集胞藻PCC 6803是光合研究中常用的模式生物。其易于转化的特性使得大量集胞藻突变体得以构建,包括许多光合功能缺陷型突变体。然而,很明显,集胞藻“野生型”亚菌株存在一系列具有明显不同表型的菌株。在这里,我们分析了一种标准的运动型巴斯德收集菌株PCC以及两种非运动型耐葡萄糖亚菌株(这里命名为GT-P和GT-W)中光合膜复合物的组织情况,这两种亚菌株之前被用作构建许多光合定点突变体的遗传背景。虽然GT-P和GT-W菌株都源自1988年由威廉姆斯构建和描述的同一菌株,但只有GT-P在色素沉着以及光系统II(PSII)和光系统I(PSI)复合物的组成上与PCC相似。相比之下,GT-W含有更多的类胡萝卜素,但叶绿素(Chl)含量显著降低,这反映在二聚体PSII尤其是三聚体PSI的水平较低上。我们发现GT-W在Chl生物合成方面存在缺陷,并且未组装的D1-D2反应中心、CP47尤其是CP43的水平异常高。GT-W的另一个特定特征是之前推测参与Chl分子循环的Ycf39-Hlip复合物水平增加了几倍。基因组重测序显示,GT-W的表型与染色体上一个大区域的串联重复有关,该区域包含100个基因,包括编码D1、Psb28和其他PSII相关蛋白以及镁原卟啉甲酯环化酶(Cycl)的基因。有趣的是,在光合自养条件下将GT-W细胞在琼脂平板上培养几个月后,这种重复完全消除了。没有重复的GT-W菌株在PSII组装方面没有明显缺陷,并且与GT-P亚菌株相似。虽然我们并不确切知道这种重复是如何影响GT-W表型的,但我们推测重复区域编码的PSII和Chl生物合成机制的蛋白质成分化学计量的改变损害了这些多亚基复合物的正确组装和功能。该研究还强调了合适的对照菌株对于评估集胞藻突变体的至关重要性。