LIFEPLAN: A worldwide biodiversity sampling design

For cases where the mobile application fails or metadata needs to be corrected, we also have a browser version: LIFEPLAN Web Admin [45]. Because metadata can be edited after collect- ing or created after the fact, we also record the fields described in Table 1. For each sample, we thus hold spatiotemporal metadata of where and when it was placed in the field and collected. Our database has additional tables with information on the Sites, Teams, Equipment, Sample codes and Users, which we can associate with the Activity by the relevant IDs. Ethics declarations The acquisition and transport of biological samples is strictly regulated by international agree- ments. All sampling and processing of samples within LIFEPLAN adheres to both national and international legislation and agreements. To comply with the Convention on Biological Diversity, with each team contributing to LIFEPLAN we have a Material Transfer Agreement (MTA). With each nation, we have estab- lished via the ABS clearing house whether Prior Informed Consent (PIC) is required, and acquired it where necessary. The MTA and PIC define Access and Benefit Sharing clauses to provide non-monetary benefits to the data providers. This study was subject to review by European Research Council Executive Agency (ERCEA) Unit B1 –Ethics Review and Expert Management. Expected results Samples for DNA based identification Physical samples can be identified with metabarcoding, using different primers depending on the species group of interest. Malaise samples can also be individually sequenced or barcoded

and photographed. In LIFEPLAN, we metabarcode all physical samples for the COI (cyto- chrome oxidase I) region for Malaise samples or ITS2 (internal transcribed spacer 2) region for cyclone and soil samples, as well as individually COI-barcoding and bulk imaging a subset of Malaise samples. Cyclone samples are collected in 1.5 ml sterile microtubes, then freeze dried and stored at -20˚C. They usually contain a small amount of fungal spores, pollen and other particles under 1 μm in diameter. Samples may contain small insects or water, which are removed and their presence noted before analysis (Fig 7A; Table 2). Malaise samples are collected in Nalgene bottles filled with pure ethanol to euthanise and conserve the arthropods, and stored at -20 C˚ (Fig 7B). Fig 7. Physical samples collected in project LIFEPLAN. (A) Two cleaned and freeze-dried cyclone samples; an insect was removed from the tube on the left. (B) A Malaise sample collected after one week in the field. (C) A soil sample inside a glassine bag. https://doi.org/10.1371/journal.pone.0313353.g007 PLOS ONE LIFEPLAN: A worldwide biodiversity sampling design PLOS ONE | https://doi.org/10.1371/journal.pone.0313353 December 31, 2024 10 / 15