Data Sheet 7_Monitoring terrestrial rewilding with environmental DNA metabarcoding: a systematic review of current trends and recommendations.csv

IntroductionRewilding, the facilitation of self-sustaining and resilient ecosystems by restoring natural processes, is an increasingly popular conservation approach and potential solution to the biodiversity and climate crises. Outcomes of rewilding can be unpredictable, and monitoring is essential to determine whether ecosystems are recovering. Metabarcoding, particularly of environmental DNA (eDNA), is revolutionizing biodiversity monitoring and could play an important role in understanding the impacts of rewilding but has mostly been applied within aquatic systems.
MethodsThis systematic review focuses on the applications of eDNA metabarcoding in terrestrial monitoring, with additional insights from metabarcoding of bulk and ingested DNA. We examine publication trends, choice of sampling substrate and focal taxa, and investigate how well metabarcoding performs compared to other monitoring methods (e.g. camera trapping).
ResultsTerrestrial ecosystems represented a small proportion of total papers, with forests the most studied system, soil and water the most popular substrates, and vertebrates the most targeted taxa. Most studies focused on measuring species richness, and few included analyzes of functional diversity. Greater species richness was found when using multiple substrates, but few studies took this approach. Metabarcoding did not consistently outperform other methods in terms of the number of vertebrate taxa detected, and this was likely influenced by choice of marker, sampling substrate and habitat.
DiscussionOur findings indicate that metabarcoding, particularly of eDNA, has the potential to play a key role in the monitoring of terrestrial rewilding, but that further ground- truthing is needed to establish the most appropriate sampling and experimental pipelines for the target taxa and terrestrial system of interest.
Systematic Review Registrationhttps://osf.io/38w9q/?view_only=47fdab224a7a43d298eccbe578f1fcf0, identifier 38w9q.

**引言**
再野化（Rewilding）是指通过恢复自然过程，助力构建自我维持且具有恢复力的生态系统，如今已成为愈发流行的保护手段，同时也是应对生物多样性与气候危机的潜在解决方案。再野化的实施效果难以预判，因此开展监测以判断生态系统是否正在恢复至关重要。元条形码测序（Metabarcoding），尤其是针对环境DNA（eDNA）的元条形码测序，正在彻底变革生物多样性监测工作，有望在解析再野化生态效应的过程中发挥关键作用，但目前该技术的应用大多局限于水生系统。

**方法**
本系统综述聚焦环境DNA元条形码测序在陆地监测中的应用，同时纳入基于批量DNA与摄食者体内DNA的元条形码测序相关研究作为补充参考。本研究梳理了相关文献的发表趋势、采样基质与目标类群的选择偏好，并对比分析了元条形码测序相较于其他监测手段（如相机陷阱法（camera trapping））的检测效能。

**结果**
已发表文献中针对陆地生态系统的研究占比偏低，其中森林是研究最多的生态系统类型，土壤与水体是最常用的采样基质，脊椎动物则是最受关注的目标类群。多数研究仅聚焦于物种丰富度的测定，极少有研究涉及功能多样性分析。采用多基质采样的研究可检测到更高的物种丰富度，但此类研究数量极少。元条形码测序在检测脊椎动物类群数量方面并未始终优于其他监测手段，其检测效能可能受到分子标记选择、采样基质与生境类型的影响。

**讨论**
本研究结果表明，元条形码测序（尤其是基于环境DNA的元条形码测序）有望在陆地再野化监测中发挥关键作用，但仍需开展进一步的实地验证，以针对目标类群与研究关注的陆地生态系统确立最适配的采样方案与实验流程。

**系统综述注册信息**：https://osf.io/38w9q/?view_only=47fdab224a7a43d298eccbe578f1fcf0，注册标识符为38w9q。