嗜盐古菌脂质生物标志物特征及其环境生态指示意义

古菌的细胞膜主要由甘油类异戊二烯醚类化合物组成。在环境中古菌脂质的组成特征能够反映群落和生境的变化，具有指示生态的能力。目前，古菌脂质研究中化合物的鉴定仍是瓶颈问题，并且组学尺度的古菌脂质研究仍处于起步阶段。本研究采用高分辨质谱结合数据库的脂质鉴定技术，以七株纯培养嗜盐古菌为研究对象，系统地探究了嗜盐古菌的脂质组构成以及各脂质类型及其对环境因子的响应特征；通过对青藏高原盐湖的脂质组分析，进一步发掘出嗜盐古菌脂质在自然环境中的生态指示意义。

相同培养条件的实验结果表明，不同嗜盐古菌产生的脂质类型和组成存在显著差异。例如，Haloferax属的菌株产生较多不饱和二醚，Natrialbales目的菌株产生大量带延长链的二醚。对脂质组进行聚类分析能够在目分类水平上区分各类嗜盐古菌，说明脂质组具有化学分类的潜力。

不同培养条件的实验结果表明，温度和盐度变化能够影响嗜盐古菌的部分脂质化合物的合成，且在不同菌株中脂质对环境因子变化的响应方式不同。例如，不饱和磷脂在Haloterrigena turkmenica中与盐度呈正相关关系，而在Haloferax larsenii中则呈负相关。然而，结果表明，环境因素对嗜盐古菌脂质组的影响有限，难以逾越菌株自身的差异。

对青藏高原盐湖沉积物的脂质组分析表明，低盐度样品中的古菌二醚可能存在多种非嗜盐古菌的生物源；高盐度样本中，嗜盐古菌来源的脂质具有更高的比例。这说明在不同盐度环境中，古菌的生态地位发生了转变。基于特征脂质在环境中的比例变化，我们构建了DER（Diether ratio）、MK87R（MK_8:7 ratio）和MK8R（MK₈ ratio）指标。它们与环境盐度存在显著的正相关，具有指示环境盐度变化的潜力。

本研究通过对纯培养菌株与盐湖沉积物的脂质组分析，拓展了对嗜盐古菌脂质组的认识，揭示了古菌脂质在青藏高原盐湖中的分布特征。在此基础上，本研究构建了三个基于古菌脂质的盐度指标，探究了古菌脂质在盐湖中的环境和生态指示意义，为利用古菌脂质组研究高盐环境的微生物生态变迁提供理论基础。

The cell membranes of archaea are predominantly composed of isoprenoid glycerol ether lipids. The compositional changes in archaeal lipids from the environment can reflect community structure and habitat variations, thus revealing ecological dynamics. Currently, lipid identification remains a bottleneck in archaeal lipid research. Archaeal lipidomics has potential to help identify novel lipid biomarkers, but this technology is still in its infancy and so far a comprehensive database of archaeal lipids does not yet exist. In this study, seven Halobacteria strains were selected as the subjects of investigation. We employed high-resolution mass spectrometry in conjunction with a database-based lipid identification method to systematically analyze the lipidome composition and its responses to environmental factors. By examining the lipidome of salt lakes on the Qinghai-Tibet Plateau (QTP), we further elucidated the ecological significance of halobacterial lipids in their natural environment.

The results revealed significant differences in the lipids produced by different strains of Halobacteria under identical culture condition. Various groups of Halobacteria exhibited distinct lipid profiles. For instance, Haloferax displayed a higher proportion of unsaturated diethers, whereas Natrialbales produced abundant diethers with extended chains, which could serve as indicators of Halobacteria taxonomy. Cluster analysis of the lipidome enabled differentiation of Halobacteria at the Order level, underscoring the chemotaxonomic potential of the halobacterial lipidome.

Temperature and salinity fluctuations were found to influence the production of specific lipids in Halobacteria, with lipids responding differently to environmental stimuli across different strains. For instance, unsaturated phospholipids exhibited a positive correlation with salinity in the pure culture of Haloterrigena turkmenica but a negative correlation in that of Haloferax larsenii. However, the impact of environmental factors on the halobacterial lipidome was relatively limited. Both clustering and principal component analysis (PCA) indicated that environmental changes had minimal influence on the lipidome of Halobacteria compared to interspecies differences.

In the QTP salt lakes with lower salinity levels, archaeal diethers from non-halophilic archaea could contribute to the lipid pool, whereas lipids associated with Halobacteria exhibited significantly higher abundance in high-salinity samples. This suggests a shift in ecological dynamics among archaea under varying salinity conditions. Based on changes in diether and quinone MK content in the environment, we developed DER (Diether ratio), MK87R (MK_8:7 ratio) and MK8R (MK₈ ratio) proxies, which show positive correlations with environmental salinity, thus representing potential candidates for reconstructing environmental salinity levels.

Overall, this study expanded our understanding of the halobacterial lipidome, delineated the distribution of archaeal lipids in QTP salt lakes, established salinity indicators utilizing archaeal lipids, and underscored the environmental and ecological significance of archaeal lipids in saline ecosystems.

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