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# Microclimate is a strong predictor of the native and invasive plant-associated soil microbiota on San Cristobal Island, Galapagos archipelago Alexi A. Schoenborn*,1, Sarah M. Yannarell*,1, Caroline T. MacVicar2, Noelia N. Barriga-Medina5,6, Meng Markillie7, Hugh Mitchell7, Kevin S. Bonham2, Antonio Leon-Reyes5,6, Diego Riveros-Iregui3,4,5, Vanja Klepac-Ceraj2,#, Elizabeth A. Shank8,# #### Here you can find QIIME2 pipeline we used to process 16S rRNA gene amplcions (bacteria) and ITS amplicons (fungi) and R-code we used to analyze and visualize the data. In addition to 16S rRNA gene data, we also looked at 6 metagenomes. ### ABSTRACT/SUMMARY Understanding the drivers that affect soil bacterial and fungal communities is essential to understanding and mitigating the impacts of human activity on vulnerable ecosystems like those on the Galapagos Islands. Located far off the coast of Ecuador, the volcanic slopes of the Galapagos Islands lead to steep elevation gradients that generate distinct microclimates across small spatial scales. Although much is known about the impacts of climate change and invasive plant species on the above-ground biodiversity of the Galapagos Islands, little is known about their resident soil microbial communities and the drivers that shape these communities. Here, we investigate the bacterial and fungal soil communities associated with invasive plant species (Psidium guajava (guava)) and native plant species across three distinct microclimates on San Cristobal Island: Mirador (arid), Cerro Alto (transition zone), and El Junco (humid). At each site, we collected soil at three depths (rhizosphere, 5 cm, and 15 cm). Sampling location was the strongest driver of both bacterial and fungal communities, with additional minor but significant impacts from plant type and soil depth. This study highlights the continued need to explore microbial communities across diverse environments and demonstrates how both abiotic and biotic factors impact soil microbial communities in the Galapagos archipelago. ### IMPORTANCE/SIGNIFICANCE Human activity (climate change, introduction of invasive species, etc.) poses a huge threat to biodiverse environments, and soil microbiota are an essential component to maintaining healthy ecosystems. The Galapagos Islands are a unique real-world laboratory, in that the islands’ volcanic geography and physical location in the Pacific Ocean generate distinct microclimates over small geographic distances. Here we explored how both bacterial and fungal soil community composition are affected by microclimate (sampling location), plant species (native or invasive plants), and soil depth with high sample replication in these young and pristine soils.  Figure 1. Within a small spatial scale, San Cristobal Island, Galapagos Archipelago, experiences distinct microclimates. (A) The Galapagos Islands are a series of islands formed from Volcanic activity. Our three field sites were located on the oldest island, San Cristobal, and had weather stations installed in 2015; Mirador (arid site - 320 m), Cerro Alto (semi-arid - 520 m) and El Junco (humid - 690 m). (B) Weather data; temperature, humidity and precipitation collected and monitored since 2015 to present, R = rainy season (January - June), D = dry season (July - December), data are the average of collections for the given season. Due to remote locations, some weather station data is absent (missing line). (C) table detailing the microclimates and different plant species we collected soil from at each site. (D) Image relaying our soil sampling scheme per plant (i) rhizosphere (root associated soil), (ii) 5 cm below the plant, and (iii) 15-20 cm below the plant, totalling 3 soil samples per plant.

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