BUSCADOR RECOLECTA

[Methods] Experiment locations and climate Trans-Mediterranean translocation of Posidonia oceanica fragments took place between Catalunya (Spain), Mallorca (Spain) and Cyprus in July 2018 and were monitored until July 2019 (Fig. 1). Sea surface temperature data for each transplant site were based on daily SST maps with a spatial resolution of 1/4°, obtained from the National Center for Environmental Information (NCEI, https://www.ncdc.noaa.gov/oisst ) (Reynolds et al. 2007). These maps have been generated through the optimal interpolation of Advanced Very High Resolution Radiometer (AVHRR) data for the period 1981-2019. Underwater temperature loggers (ONSET Hobo pro v2 Data logger) were deployed at the transplant sites in Catalunya, Mallorca and Cyprus and recorded hourly temperatures throughout the duration of the experiment (one year). In order to obtain an extended time series of temperature at transplant sites, a calibration procedure was performed comparing logger data with sea surface temperature from the nearest point on SST maps. In particular, SST data were linearly fitted to logger data for the common period. Then, the calibration coefficients were applied to the whole SST time series to obtain corrected-SST data and reconstruct daily habitat temperatures from 1981-2019. Local climate data was also compared to the global thermal distribution of P. oceanica to assess how representative experimental sites were of the thermal distribution of the species (Supplementary materials). Collectively, seawater temperatures from the three locations span the 16th - 99th percentile of temperatures observed across the global thermal distribution of P. oceanica. As such Catalunya, Mallorca and Cyprus are herein considered to represent the cool-edge, centre and warm-edge of P. oceanica distribution, respectively. Transplantation took place toward warmer climates and procedural controls were conducted within each source location, resulting in six source-to-recipient combinations (i.e. treatments, Fig. 1). Initial collection of P. oceanica, handling and transplantation was carried out simultaneously by coordinated teams in July 2018 (Table S1). Each recipient location was subsequently resampled four times over the course of the experiment, in August/September 2018 (T1), October 2018 (T2), April 2019 (T3) and May/June 2019 (T4, Table S1). Between 60-100 fragments were collected for each treatment. A fragment was defined as a section of P. oceanica containing one apical shoot connected with approximately five vertical shoots by approximately 10-15 cm of rhizome with intact roots. Collection occurred at two sites within each location, separated by approximately 1 km. Within sites, collections were conducted between 4 – 5 m depth and were spaced across the meadow to minimise the dominance of a single clone and damage to the meadow. Upon collection, fragments were transported for up to one hour back to the nearest laboratory in shaded seawater. Handling methods In the laboratory, fragments were placed into holding tanks with aerated seawater, at ambient temperature and a 14:10 light-dark cycle. All shoots were clipped to 25 cm length (from meristem to the tip of the longest leaves), to standardise initial conditions and reduce biomass for transportation. For transport by plane or ferry between locations, fragments were packed in layers within cool-boxes. Each layer was separated by frozen cool-packs wrapped in wet tea towels (rinsed in sea water). All fragments spent 12 hrs inside a cool-box irrespective of their recipient destination, including procedural controls (i.e. cool-cool, centre-centre and warm-warm) to simulate the transit times of the plants travelling furthest from their source location (Fig. 1a). On arrival at the destination, fragments were placed in holding tanks with aerated seawater at ambient temperature as described above in their recipient location for 48 hrs, prior to field transplantation. Measurement methods One day prior to transplantation, fragments were tagged with a unique number and attached to U-shaped peg with cable-ties. Morphological traits for each fragment were measured and included: 1) length of the longest apical leaf, width and number of leaves 2) total number of bite marks on leaves of three vertical shoots per fragment, 3) number of vertical shoots, 4) leaf count of three vertical shoots per fragment and 5) overall horizontal rhizome length. A subset (n=10) of fragments per treatment were marked prior transplantation to measure shoot growth. To do this, all shoots within a single fragment were pierced using a hypodermic needle. Two holes were pierced side-by-side at the base of the leaf/top of the meristem. Transplant methods All transplant sites were located in 4 – 5 m depth in area of open dead-matte, surrounded by P. oceanica meadow. In Mallorca and Cyprus, fragments were distributed between two sites, separated by approximately 1 km. In Catalunya, a lack of suitable dead matte habitat, meant that all fragments were placed in one site. Fragments were planted along parallel transects at 50 cm intervals and with a 50 cm gap between parallel transects (Fig. S1). Different treatments were mixed and deployed haphazardly along each transect. Resampling methods and herbivory On day 10 of the experiment, a severe herbivory event was recorded at both warm-edge translocation sites. Scaled photos of all fragments were taken at this time to record the effects of herbivory on transplants. At the end of each main sampling period (T0 – T1, T1-T2 and T3 – T4), all pierced fragments were collected and taken back to the laboratory to measure shoot growth. At T1, T2 and T3, additional sets of fragments (n = 10 per treatment) were marked using the piercing method to record growth in the subsequent time period. In addition, at T1 and T3, n = 20 shoots within the natural meadow at each site were marked to compare growth rates between the native meadow and transplants. Underwater shoot counts and a scaled photo was taken to record fragment survivorship, shoot mortality, bite marks, and shoot length among all remaining fragments within each site and sampling time. In the laboratory, morphological measurements (described above) were repeated on the collected fragments and growth of transplant and natural meadow shoots was measured. Growth (shoot elongation, cm d-1) of the marked shoots was obtained by measuring the length from the base of meristem to marked holes of each leaf (new growth) of the shoot and dividing the leaf elongation per shoot by the marking period (in days). For each shoot, total leaf length (cm shoot-1) and the number of new leaves was also recorded. The rate of new leaf production (new leaves shoot-1 d-1) was estimated dividing the number of new leaves produced per shoot and the marking period. New growth was dried at 60 ºC for 48 hrs to determine carbon and nitrogen content of the leaves, and carbon to nitrogen (C:N) ratios. Carbon and nitrogen concentrations in the new growth leaf tissue was measured at the beginning of the experiment and each subsequent time point for each treatment. Nutrient analyses were conducted at Unidade de Técnicas Instrumentais de Análise (University of Coruña, Spain) with an elemental analyser FlashEA112 (ThermoFinnigan). Underwater photos of shoots were analysed using ImageJ software (https://imagej.net). Maximum leaf length on each shoot in warm-edge transplant sites (cool-warm, centre-warm and warm-warm) were recorded for the initial (day 10) herbivore impact, T1, T2 and T3 time-points and related to transplant nutrient concentrations. Herbivore impact was estimated as the proportional change in length of the longest leaf relative to initial length at T0. Thermal stress Long term maximum temperatures were recorded as the average of annual maximum daily temperatures in each transplant site, averaged between years from 1981-2019. Maximum thermal anomalies were calculated as the difference between daily temperatures in a recipient site over the course of the experiment and the long-term maximum temperature in the source site for each corresponding population. ‘Heat stress’ and ‘recovery’ growth periods of the experiment were defined as T0 -T2 (July-October) and T2-T4 (November-June), respectively, corresponding to periods of positive and negative maximum thermal anomalies. Thermal anomalies experienced by the different transplant treatments were plotted using the ‘geom_flame’, function in the ‘HeatwavesR’ package (Schlegel & Smit 2018) of R (version 3.6.1, 2019) ., 1. The prevalence of local adaptation and phenotypic plasticity among populations is critical to accurately predicting when and where climate change impacts will occur. Currently, comparisons of thermal performance between populations are untested for most marine species or overlooked by models predicting the thermal sensitivity of species to extirpation. 2. Here we compared the ecological response and recovery of seagrass populations (Posidonia oceanica) to thermal stress throughout a year-long translocation experiment across a 2800 km gradient in ocean climate. Transplants in central and warm-edge locations experienced temperatures >29 ºC, representing thermal anomalies >5ºC above long-term maxima for cool-edge populations, 1.5ºC for central and <1ºC for warm-edge populations. 3. Cool, central and warm-edge populations differed in thermal performance when grown under common conditions, but patterns contrasted with expectations based on thermal geography. Cool-edge populations did not differ from warm-edge populations under common conditions and performed significantly better than central populations in growth and survival. 4. Our findings reveal that thermal performance does not necessarily reflect the thermal geography of a species. We demonstrate that warm-edge populations can be less sensitive to thermal stress than cooler, central populations suggesting that Mediterranean seagrasses have greater resilience to warming than current paradigms suggest., Australian Research Council, Award: DE200100900. Horizon 2020 Framework Programme, Award: 659246. Fundación BBVA., Peer reviewed

1 figure, Including the identification of known and putative novel miRNAs, miRNA abundance profiling and differential abundance analysis. rRNA: ribosomal RNA; tRNA: transfer RNA; snoRNA: small nucleolar RNA; snRNA: small nuclear RNA; RE: repeat elements; qPCR: quantitative real-time PCR., Peer reviewed

1 figure, CKD: Chronic kidney disease; PN: Pyelonephritis; SB/C: Subclinical bacteriuria/Cystitis; UO: Ureteral obstruction, Peer reviewed

A. PCA of urine samples on the basis of normalized read counts of the known and putative novel miRNAs for the 38 samples initially processed. The red arrows indicate the outlier Control samples (C5, C6 and C7). B. PCA excluding the high outlier samples. CKD: Chronic kidney disease; PN: Pyelonephritis; SB/C: Subclinical bacteriuria/Cystitis; UO: Ureteral obstruction., Peer reviewed

A. Proportion of samples for which each of the known miRNAs across the different groups were detected. B. Cumulative abundance of the known feline miRNAs. The dots indicate the log10 of the miRNA abundance for each miRNA. miRNAs are sorted in each group in a decreasing order by their miRNA abundance on the x-axis, independently for each group. C. Proportion of samples for which each of the putative novel miRNA candidates across the different groups were detected. D. Cumulative abundance of the putative novel miRNAs. The dots indicate the log10 of the miRNA abundance for each miRNA. miRNAs are sorted in each group in a decreasing order by their miRNA abundance on the x-axis, independently for each group. CKD: Chronic kidney disease; PN: Pyelonephritis; SB/C: Subclinical bacteriuria/Cystitis; UO: Ureteral obstruction, CPM: Counts per million., Peer reviewed

1 figure., All samples together (all groups), as well as each one of the contrasts considered (Controls vs. PN; Control vs. SB/C; Control vs. UO; Control vs. CKD; PN vs. SB/C; PN vs. UO; PN vs. CKD and PN vs. other Pathologies) are shown. CKD: Chronic kidney disease; PN: Pyelonephritis; SB/C: Subclinical bacteriuria/Cystitis; UO: Ureteral obstruction., Peer reviewed

1 figure., CKD: Chronic kidney disease; PN: Pyelonephritis; SB/C: Subclinical bacteriuria/Cystitis; UO: Ureteral obstruction, CPM: Counts per million, Rq: Relative quantities., Peer reviewed

The data presented is from selected miRNAs that were DA (|log2FC| ≥ 1.5 for qPCR and |log2FC| ≥ 2 for small RNAseq; q-value < 0.05) using both methodologies. CKD: Chronic kidney disease; PN: Pyelonephritis; SB/C: Subclinical bacteriuria/Cystitis; UO: Ureteral obstruction, CPM: Counts per million, Rq: Relative quantities., Peer reviewed

1 table., The table includes for each miRNA the arguments for its selection for further validation, the forward and reverse sequence, the miRBase sequence used as template for primer design, if successful miRNA amplification was obtained with qPCR and qPCR amplification efficiency., Peer reviewed

1 table, Total RNA was isolated from whole urine of 38 cats, and 6 μl of each RNA sample was applied in small RNAseq. Concentration and ratios were obtained using Nanodrop measurement., Peer reviewed