Frébortová Jitka, Plíhal Ondřej, Florová Vendula, Kokáš Filip, Kubiasová Karolina, Greplová Marta, Šimura Jan, Novák Ondřej, Frébort Ivo
Department of Chemical Biology and Genetics, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Šlechtitelů 27, 783 71, Olomouc, Czech Republic.
Department of Molecular Biology, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Šlechtitelů 27, 783 71, Olomouc, Czech Republic.
J Phycol. 2017 Jun;53(3):703-714. doi: 10.1111/jpy.12538. Epub 2017 Apr 26.
Cytokinins are an important group of plant hormones that are also found in other organisms, including cyanobacteria. While various aspects of cytokinin function and metabolism are well understood in plants, the information is limited for cyanobacteria. In this study, we first experimentally confirmed a prenylation of tRNA by recombinant isopentenyl transferase NoIPT2 from Nostoc sp. PCC 7120, whose encoding gene we previously identified in Nostoc genome along with the gene for adenylate isopentenyl transferase NoIPT1. In contrast to NoIPT2, the transcription of NoIPT1 was strongly activated during the dark period and was followed by an increase in the cytokinin content several hours later in the light period. Dominant cytokinin metabolites detected at all time points were free bases and monophosphates of isopentenyladenine and cis-zeatin, while N-glucosides were not detected at all. Whole transcriptome differential expression analysis of cultures of the above Nostoc strain treated by cytokinin compared to untreated controls indicated that cytokinin together with light trigger expression of several genes related to signal transduction, including two-component sensor histidine kinases and two-component hybrid sensors and regulators. One of the affected histidine kinases with a cyclase/histidine kinase-associated sensory extracellular domain similar to the cytokinin-binding domain in plant cytokinin receptors was able to modestly bind isopentenyladenine. The data show that the genetic disposition allows Nostoc not only to produce free cytokinins and prenylate tRNA but also modulate the cytokinin biosynthesis in response to light, triggering complex changes in sensing and regulation.
细胞分裂素是一类重要的植物激素,在包括蓝细菌在内的其他生物体中也有发现。虽然植物中细胞分裂素的功能和代谢的各个方面已得到充分了解,但关于蓝细菌的相关信息却很有限。在本研究中,我们首先通过实验证实了来自 Nostoc sp. PCC 7120 的重组异戊烯基转移酶 NoIPT2 对 tRNA 的异戊烯基化作用,我们之前在 Nostoc 基因组中鉴定出了其编码基因以及腺苷酸异戊烯基转移酶 NoIPT1 的基因。与 NoIPT2 不同,NoIPT1 的转录在黑暗期被强烈激活,随后在光照期数小时后细胞分裂素含量增加。在所有时间点检测到的主要细胞分裂素代谢产物是异戊烯基腺嘌呤和顺式玉米素的游离碱和单磷酸盐,而根本未检测到 N - 糖苷。与未处理的对照相比,对上述 Nostoc 菌株用细胞分裂素处理后的培养物进行全转录组差异表达分析表明,细胞分裂素与光共同触发了几个与信号转导相关的基因的表达,包括双组分传感器组氨酸激酶以及双组分混合传感器和调节因子。其中一种受影响的组氨酸激酶具有与植物细胞分裂素受体中的细胞分裂素结合域相似的环化酶/组氨酸激酶相关的传感细胞外结构域,能够适度结合异戊烯基腺嘌呤。数据表明,这种遗传特性使 Nostoc 不仅能够产生游离细胞分裂素并对 tRNA 进行异戊烯基化,还能响应光照调节细胞分裂素的生物合成,引发传感和调节方面的复杂变化。