青藏高原高寒草甸土壤病毒富集方法优化

Optimization of Soil Virus Enrichment Method in Alpine Meadows of the Qinghai-Tibet Plateau

摘要

病毒广泛分布于土壤，通过调控宿主群落和驱动元素循环深刻影响着生物地球化学循环，包括污染物降解等对环境污染及其修复有重要价值的功能过程。青藏高原是我国的生态安全屏障，其高寒草甸土壤因草毡层植物根系致密、腐殖质层较厚等特征，导致传统土壤病毒富集方法面临着杂质干扰严重、洗脱效率不高等挑战。针对上述问题，本研究针对青藏高原草甸土壤系统优化了病毒富集流程：通过引入KPEG-M洗脱液促进了病毒解吸，采用两步过滤策略缓解了滤膜堵塞，利用离心超滤管提升了浓缩效率。与针对北美温带森林泥炭地与中国亚热带农田等土壤提出的传统病毒富集方案相比，优化体系显著提高了 DNA 回收量（P<0.05）达 11.25%~44.30%，且浓缩阶段操作便捷性更优。该优化体系的构建为解析高寒土壤病毒群落的组成、多样性与功能提供了支撑。

Abstract

Viruses are widely distributed in soils, profoundly affect biogeochemical cycles by regulating host communities and driving element cycles, including functional processes such as pollutant degradation that are of great value to environmental pollution and its restoration. The Qinghai-Tibet Plateau is an ecological security barrier in China. Its alpine meadow soils have dense roots of grass layers and thick humus layers, which leads to challenges such as severe impurity interference and low elution efficiency in traditional soil virus enrichment methods. In response to the above problems, this study optimized the virus enrichment process for the Qinghai-Tibet Plateau meadow soil system: the introduction of KPEG-M eluent promoted virus desorption, the two-step filtration strategy alleviated the clogging of the filter membrane, and the centrifugal ultrafiltration tube was used to improve the concentration efficiency. Compared with the traditional virus enrichment schemes proposed for soils such as temperate forest peatlands in North America and subtropical farmlands in China, the optimized system significantly increased the DNA recovery (P<0.05) by 11.25%~44.30%, and the concentration stage was more convenient to operate. The construction of this optimized system provides support for the analysis of the composition, diversity and function of alpine soil virus communities.

关键词

土壤病毒 / 富集方法 / 青藏高原 / 高寒草甸土壤

Key words

soil viruses / viral enrichment / Qinghai-Tibet Plateau / alpine meadow soil

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