Molecular adaptation of ammonia monooxygenase during independent pH specialization in Thaumarchaeota

Microbes are abundant in nature and often highly adapted to local conditions. While great progress has been made in understanding the ecological factors driving their distribution in complex environments, the underpinning molecular‐evolutionary mechanisms are rarely dissected. Therefore, we scrutinized the coupling of environmental and molecular adaptation in Thaumarchaeota, an abundant archaeal phylum with a key role in ammonia oxidation. These microbes are adapted to a diverse spectrum of environmental conditions, with pH being a key factor shaping their contemporary distribution and evolutionary diversification. We integrated high‐throughput sequencing data spanning a broad representation of ammonia‐oxidizing terrestrial lineages with codon modelling analyses, testing the hypothesis that ammonia monooxygenase subunit A (AmoA) – a highly conserved membrane protein crucial for ammonia oxidation and classical marker in microbial ecology – underwent adaptation during specialization to extreme pH environments. While purifying selection has been an important factor limiting AmoA evolution, we identified episodic shifts in selective pressure at the base of two phylogenetically distant lineages that independently adapted to acidic conditions and subsequently gained lasting ecological success. This involved nonconvergent selective mechanisms (positive selection vs. selection acting on variants fixed during an episode of relaxed selection) leading to unique sets of amino acid substitutions that remained fixed across the radiation of both acidophilic lineages, highlighting persistent adaptive value in acidic environments. Our data demonstrates distinct trajectories of AmoA evolution despite convergent phenotypic adaptation, suggesting that microbial environmental specialization can be associated with diverse signals of molecular adaptation, even for marker genes employed routinely by microbial ecologists.     

珠江口外东澳岛海域MG I古菌和藻类的昼夜变化关系的研究

【目的】海洋古菌MG I (marine group I archaea)是海洋中主要微生物类群,拥有利用氨氮进行氨氧化自养的能力,是海洋环境中氨氧化过程的重要参与者。研究MG I古菌的昼夜变化规律,对揭示海洋中氨氧化过程以及碳氮循环有着重要的意义。【方法】此次研究的样品来自于珠江口东澳岛附近海域,使用无人机采样技术获取了以2 h为间隔的22 h连续时间序列海水样品。本研究重点关注以下科学问题：(1)昼夜尺度下珠江口MG I古菌与藻类的群落和丰度变化特征;(2)昼夜尺度下珠江口MG I古菌受温度和藻类的影响。本研究通过样品DNA,以qPCR、二代基因测序等手段,结合环境参数(温度、盐度、营养盐浓度等),以探究海水藻类与MG I古菌之间可能存在的关系。【结果】研究发现,MG I古菌丰度为(9.1±3.2)×107 copies/L,藻类的丰度为(3.7±0.7)×108 copies/L。通过古菌的高通量测序发现MG I古菌是该区域最主要的古菌类群(36.2%–50.0%)。在昼夜变化尺度下,MG I古菌与藻类的丰度表现出一定的负相关关系,且环境因子中,温度与MG I古菌之间表现出显著的...     

Dynamics of a bloom of halophilic archaea in the Dead Sea

After a period of more than ten years in which bacterial and algal community sizes were extremely small, a dense bloom of halophilic archaea developed in the upper 5–10 m of the Dead Sea water column in the summer of 1992. The development of this bloom followed a dilution of the upper water layer by winter rainfloods, which enabled the development of a short-lived dense bloom of the unicellular green alga Dunaliella parva. The dense archaeal community (up to 3.5 × 10⁷ cells m1^–1 in June 1992) imparted a red coloration to the Dead Sea, due to its high content of bacterioruberin. Bacteriorhodopsin was not detected. High levels of potential heterotrophic activity were associated with the bloom, as measured by the incorporation of labeled organic substrates. After the decline of the algal bloom, archaeal numbers in the lake decreased only little, and most of the community was still present at the end of 1993. The amount of carotenoid pigment per cell, however, decreased 2–3-fold between June 1992 and August 1993. No new algal and archaeal blooms developed after the winter floods of 1992–1993, in spite of the fact that salinity values in the surface layer were sufficiently low to support a new algal bloom. A remnant of the 1992 Dunaliella bloom maintained itself at the lower end of the pycnocline at depths between 7 and 13 m (September 1992–August 1993). Its photosynthetic activity was small, and very little stimulation of archaeal growth and activity was associated with this algal community.     

人工湿地氮去除关键功能微生物生态学研究进展

人工湿地是一种能有效处理水体氮素污染的生态技术,其中微生物是驱动人工湿地系统中氮素去除的重要引擎。近20年来,随着分子生物学技术的广泛应用,有关人工湿地氮去除功能微生物生态学方面研究取得了一些重要进展。以硝化-反硝化作用和厌氧氨氧化作用这两种重要的人工湿地微生物脱氮途径为主,针对氨氧化细菌/古菌、厌氧氨氧化菌和反硝化菌等关键脱氮功能微生物的研究,重点归纳总结了目前有关这几类关键功能菌群在人工湿地中的丰度、活性、多样性、分布特征与影响因素,及其对废水中氮去除的作用,并在此基础上对今后的重点研究工作提出了展望。面向未来人工湿地氮去除关键功能微生物的研究应侧重其在污水净化和温室气体减排等方面的生态功能研究,同时加强其代谢过程与机制以及不同功能菌群间的关联研究。     

极端嗜盐古生菌(Natrinema sp.)R6-5胞外嗜盐蛋白酶的纯化和性质研究

采用bacitracin-Sepharose 4B亲和层析的方法得到凝胶电泳均一的来自极端嗜盐古生菌(Natrinema sp.)R6-5的胞外嗜盐蛋白酶。经SDS-PAGE分析该酶亚基分子量为62kDa。PMSF对它的活性完全抑制,表明它是一种丝氨酸蛋白酶,该酶反应的最适NaCl浓度为3mol/L,最适温度为45℃,最适pH值为8.0。在高盐条件下能维持高活性并十分稳定,具有重要的潜在应用价值。     

Abnormal junctional membrane structures in cardiac myocytes expressing ectopic junctophilin type 1

A gene encoding a putative ATP-dependent DNA ligase from the aerobic hyperthermophilic archaeon Aeropyrum pernix K1 was cloned and the biochemical characteristics of the resulting recombinant protein were examined. The gene (accession no. APE1094) from A. pernix encoding a 69-kDa protein showed a 39–61% identity with other ATP-dependent DNA ligases from the archaea. Normally DNA ligase is activated by NAD⁺ or ATP. There has been no report about the other activators for DNA ligase. The recombinant ligase was a monomeric protein and catalyzed strand joining on a singly nicked DNA substrate in the presence of ADP and a divalent cation (Mg²⁺, Mn²⁺, Ca²⁺ and Co²⁺) at high temperature. The optimum temperature and pH for nick-closing activity were above 70°C and 7.5°C, respectively. The ligase remained stable for 60 min of treatment at 100°C, and the half-life was about 25 min at 110°C. This is the first report of a novel hyperthermostable DNA ligase that can utilize ADP to activate the enzyme.     

Isolation and Phylogenetic Analysis of Halophilic Archaeon AJ6

Halophilic archaeon AJ6 was isolated and purified from the Altun Mountain National Nature Reserve of the Xinjiang Uygur Autonomous Region. Strain AJ6 is a Gram-negative rod whose size is 0.2–0.6 by 1.6–4.2 μm, wherein a few cells are globular. The optimum salt concentration for its growth is 20% NaCl and 0.6% Mg²⁺, and the optimum pH is 6.0–7.0. Morphological, physiological, and biochemical characteristics of strain AJ6 were observed. The 16S rRNA encoding gene (16S rDNA) sequence of strain AJ6 was amplified by PCR, and its nucleotide sequence was determined subsequently. “Clustalw” and “PHYLIP” software bags were used to analyze the 16S rDNA sequence; the homology was compared, and then the phylogenetic tree was established. The results indicate that strain AJ6 is a novel species of the genus Natrinema. The GenBank accession number of the 16S rDNA sequences of strain AJ6 is AY277584. Translated from Journal of Zhejiang University (Science Edition), 2005, 32(1) (in Chinese)     

极端嗜热古菌DNA修复核酸内切酶的研究进展

极端嗜热古菌由于生活在高温环境,其基因组DNA面临着严重的挑战,因此,它们如何维持其基因组稳定是本研究领域最为关注的科学问题之一。极端嗜热古菌具有与常温微生物相似的自发突变频率,暗示着它们比常温微生物具有更加有效的DNA修复体系进行修复高温所造成的基因组DNA损伤。目前,极端嗜热古菌DNA修复的分子机制尚不清楚。核酸内切酶在DNA修复途径中发挥着重要的作用。基因组序列显示极端嗜热古菌编码多种DNA修复核酸内切酶,但是其研究尚处于初期阶段。本文综述了极端嗜热古菌DNA修复核酸内切酶Nuc S、Endo V、Endo Q、XPF和Hjc的研究进展,并对今后的研究提出了展望。     

嗜盐菌耐盐机制相关基因的研究进展

嗜盐微生物能够在高盐环境中生存,其耐盐机制一直是微生物学家研究的热点。目前嗜盐微生物耐盐机制的研究主要集中在细胞吸K+排Na+作用、胞内积累小分子相容性溶质及嗜盐酶的氨基酸组成特性三个方面。文章从基因水平综述了嗜盐菌的耐盐机制,并对其在高盐废水处理上的应用进行讨论与展望。