School of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan, 430023, Hubei, China.
College of Resources and Environment, Yangtze University, Wuhan, 430100, Hubei, China.
BMC Biotechnol. 2020 Nov 30;20(1):61. doi: 10.1186/s12896-020-00656-5.
Harmful cyanobacterial blooms have attracted wide attention all over the world as they cause water quality deterioration and ecosystem health issues. Microcystis aeruginosa associated with a large number of bacteria is one of the most common and widespread bloom-forming cyanobacteria that secret toxins. These associated bacteria are considered to benefit from organic substrates released by the cyanobacterium. In order to avoid the influence of associated heterotrophic bacteria on the target cyanobacteria for physiological and molecular studies, it is urgent to obtain an axenic M. aeruginosa culture and further investigate the specific interaction between the heterotroph and the cyanobacterium.
A traditional and reliable method based on solid-liquid alternate cultivation was carried out to purify the xenic cyanobacterium M. aeruginosa FACHB-905. On the basis of 16S rDNA gene sequences, two associated bacteria named strain B905-1 and strain B905-2, were identified as Pannonibacter sp. and Chryseobacterium sp. with a 99 and 97% similarity value, respectively. The axenic M. aeruginosa FACHB-905A (Microcystis 905A) was not able to form colonies on BG agar medium without the addition of strain B905-1, while it grew well in BG liquid medium. Although the presence of B905-1 was not indispensable for the growth of Microcystis 905A, B905-1 had a positive effect on promoting the growth of Microcystis 905A.
The associated bacteria were eliminated by solid-liquid alternate cultivation method and the axenic Microcystis 905A was successfully purified. The associated bacterium B905-1 has the potentiality to promote the growth of Microcystis 905A. Moreover, the purification technique for cyanobacteria described in this study is potentially applicable to a wider range of unicellular cyanobacteria.
有害蓝藻水华已引起全世界的广泛关注,因为它们会导致水质恶化和生态系统健康问题。与大量细菌相关的铜绿微囊藻是最常见和广泛分布的形成水华的蓝藻之一,它会分泌毒素。这些相关细菌被认为受益于蓝藻释放的有机基质。为了避免相关异养细菌对目标蓝藻的生理和分子研究的影响,迫切需要获得无菌铜绿微囊藻培养物,并进一步研究异养菌与蓝藻之间的特定相互作用。
基于固液交替培养的传统可靠方法,对异养铜绿微囊藻 FACHB-905 进行了纯化。根据 16S rDNA 基因序列,鉴定出两种相关细菌,分别命名为 Pannonibacter sp. 和 Chryseobacterium sp.,相似度值分别为 99%和 97%。无菌铜绿微囊藻 FACHB-905A(微囊藻 905A)在不添加 B905-1 的 BG 琼脂培养基上无法形成菌落,而在 BG 液体培养基中生长良好。虽然 B905-1 的存在对微囊藻 905A 的生长不是必需的,但它对促进微囊藻 905A 的生长有积极的影响。
通过固液交替培养方法消除了相关细菌,并成功纯化了无菌微囊藻 905A。相关细菌 B905-1 具有促进微囊藻 905A 生长的潜力。此外,本研究中描述的蓝藻纯化技术可能适用于更广泛的单细胞蓝藻。