Alcántara-Hernández Rocio J, Valenzuela-Encinas César, Zavala-Díaz de la Serna Francisco J, Rodriguez-Revilla Javier, Dendooven Luc, Marsch Rodolfo
Departamento de Biotecnología y Bioingeniería, GIB Cinvestav, México, D.F., Mexico.
FEMS Microbiol Lett. 2009 Sep;298(1):56-66. doi: 10.1111/j.1574-6968.2009.01710.x.
Assimilatory nitrate reduction (ANR) is a pathway wherein NO(3)(-) is reduced to NH(4)(+), an N species that can be incorporated into the biomass. There is little information about the ANR genes in Archaea and most of the known information has been obtained from cultivable species. In this study, the diversity of the haloarchaeal assimilatory nitrate-reducing community was studied in an extreme saline alkaline soil of the former lake Texcoco (Mexico). Genes coding for the assimilatory nitrate reductase (narB) and the assimilatory nitrite reductase (nirA) were used as functional markers. Primers to amplify and detect partial narB and nirA were designed. The analysis of these amplicons by cloning and sequencing showed that the deduced protein fragments shared >45% identity with other NarB and NirA proteins from Euryarchaeota and <38% identity with other nitrate reductases from Bacteria and Crenarchaeota. Furthermore, these clone sequences were clustered within the class Halobacteria with strong support values in both constructed dendrograms, confirming that desired PCR products were obtained. The metabolic capacity to assimilate nitrate by these haloarchaea seems to be important given that at pH 10 and higher, NH(4)(+) is mostly converted to toxic and volatile NH(3), and NO(3)(-) becomes the preferable N source.
同化性硝酸盐还原(ANR)是一条将NO₃⁻还原为NH₄⁺的途径,NH₄⁺是一种可被纳入生物质的氮物种。关于古菌中ANR基因的信息很少,并且大多数已知信息是从可培养物种中获得的。在本研究中,在前特斯科科湖(墨西哥)的极端盐碱土壤中研究了嗜盐古菌同化性硝酸盐还原群落的多样性。编码同化性硝酸盐还原酶(narB)和同化性亚硝酸还原酶(nirA)的基因被用作功能标记。设计了用于扩增和检测部分narB和nirA的引物。通过克隆和测序对这些扩增子进行分析表明,推导的蛋白质片段与广古菌门的其他NarB和NirA蛋白质具有>45%的同一性,与细菌和泉古菌门的其他硝酸盐还原酶具有<38%的同一性。此外,在构建的两个系统发育树中,这些克隆序列都在嗜盐菌纲内聚类,支持值很高,证实获得了所需的PCR产物。鉴于在pH值为10及更高时,NH₄⁺大多转化为有毒且易挥发的NH₃,而NO₃⁻成为更可取的氮源,这些嗜盐古菌同化硝酸盐的代谢能力似乎很重要。