Departamento de Ciencias Ambientales, Universidad Autónoma Metropolitana-Lerma, Hidalgo Pte. 46, Col. La Estación, 52006, Lerma de Villada, Estado de México, Mexico.
Grupo de Ecología Genética Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, C/ Profesor Albareda 1, 18008, Granada, Spain.
Int Microbiol. 2020 Aug;23(3):467-474. doi: 10.1007/s10123-020-00119-z. Epub 2020 Jan 14.
Treatment of environmental samples under field conditions may require the application of chemical preservatives, although their use sometimes produces changes in the microbial communities. Sodium azide, a commonly used preservative, is known to differentially affect the growth of bacteria. Application of azide and darkness incubation to Isabel soda lake water samples induced changes in the structure of the bacterial community, as assessed by partial 16S rRNA gene pyrosequencing. Untreated water samples (WU) were dominated by gammaproteobacterial sequences accounting for 86%, while in the azide-treated (WA) samples, this group was reduced to 33% abundance, and cyanobacteria-related sequences became dominant with 53%. Shotgun sequencing and genome recruitment analyses pointed to Halomonas campanensis strain LS21 (genome size 4.07 Mbp) and Synechococcus sp. RS9917 (2.58 Mbp) as the higher recruiting genomes from the sequence reads of WA and WU environmental libraries, respectively, covering nearly the complete genomes. Combined treatment of water samples with sodium azide and darkness has proven effective on the selective enrichment of a cyanobacterial group. This approach may allow the complete (or almost-complete) genome sequencing of Cyanobacteria from metagenomic DNA of different origins, and thus increasing the number of the underrepresented cyanobacterial genomes in the databases.
在野外条件下处理环境样品可能需要应用化学防腐剂,尽管它们的使用有时会导致微生物群落发生变化。叠氮化钠是一种常用的防腐剂,已知其对细菌的生长有差异影响。将叠氮化钠和黑暗孵育应用于伊莎贝尔苏打湖水样,通过部分 16S rRNA 基因焦磷酸测序评估,导致细菌群落结构发生变化。未经处理的水样(WU)主要由占 86%的γ变形菌序列组成,而在叠氮化钠处理的(WA)样品中,该组丰度减少到 33%,而与蓝细菌相关的序列变得占优势,达到 53%。鸟枪法测序和基因组招募分析表明,Halomonas campanensis 菌株 LS21(基因组大小为 4.07 Mbp)和 Synechococcus sp. RS9917(2.58 Mbp)分别是 WA 和 WU 环境文库序列中招募基因组的较高基因组,几乎涵盖了完整的基因组。水样中叠氮化钠和黑暗的联合处理已被证明可有效选择性富集蓝细菌群体。这种方法可以允许对不同来源的宏基因组 DNA 中的蓝细菌进行完整(或几乎完整)基因组测序,从而增加数据库中代表性不足的蓝细菌基因组的数量。