Phenotypic plasticity of European larch radial growth and wood density along a‐1,000 m elevational gradient

Why this research Matters Phenotypic plasticity is a key mechanism for sedentary long‐living species to adjust to changing environment. Here, we use mature Larix decidua tree‐ring variables collected along an elevational transect in the French Alps to characterize the range of individual plastic responses to temperature. Stem cores from 821 mature Larix decidua trees have been collected from four plots distributed along a 1,000‐m elevational gradient in a natural forest to build up individual linear reaction norms of tree‐ring microdensity traits to temperature. The sign, magnitude and spread of variations of the slopes of the individual reaction norms were used to characterize variation of phenotypic plasticity among plots and traits. Results showed a large range of phenotypic plasticity (with positive and negative slopes) at each elevational plot and for each tree‐ring variable. Overall, phenotypic plasticity tends to be larger but positive at higher elevation, negative at the warmer lower sites, and more variable in the center of the elevation distribution. Individual inter‐ring reaction norm is a valuable tool to retrospectively characterize phenotypic plasticity of mature forest trees. This approach applied to Larix decidua tree‐ring micro‐density traits along an elevation gradient showed the existence of large inter‐individual variations that could support local adaptation to a fast‐changing climate.

mechanism if the length of the change is much less than the generation time (Bradshaw, 1965). However, despite the accelerated interest in unravelling the genetic basis of PP in relation to the fastincreasing global warming, its quantification for the endangered sedentary and long-living organisms remains a challenge and is scarce.
PP is usually estimated using Reaction Norm (RN), which is a linear or non-linear function indicating the amount of phenotypic change of a genotype between environments (Arnold et al., 2019;Falconer, 1990;Morrissey & Liefting, 2016;Via et al., 1995).
Generally, the environmental variability is generated by exposing the same genotype to different conditions. Spatial (site-related) and temporal (time-related) PP refer to the types of environmental variation (Scheiner, 2013). The spatial PP requires reciprocal experiments and copies of the same genotype in different sites or controlled conditions (Via, 1993;Vitasse et al., 2010). The temporal PP demands repeated measurements (Araya-Ajoy et al., 2015) or retrospective evaluations (Fonti et al., 2010;Marchal et al., 2019). While the definition is simple, accurate spatial (or site-related) RN are often difficult to estimate, since the number of sites or controlled conditions is generally low and their environmental value is most of the time problematic to quantify (Pigliucci & Pigliucci, 2001). Temporal (or time-related) PP can be estimated from series of repeated measurements, or, for some organisms like fish and tree species, retrospectively using temporal markers (scales in fish and annual rings in trees; Black et al., 2019;Fallour-Rubio et al., 2009). Estimation of temporal PP is never straightforward, since it can be confused with the ontogenic development of the organism.
Trees have a high potential for recording signals at different temporal scales and frequencies. Many studies explored tree response to environmental variation based on radial growth, annual ring formation and relationships with climate estimated at the population level. In particular, dendrochronology (Büntgen, 2019;Vitas, 2018) and dendroecology (Manzanedo & Pederson, 2019;Pompa-García et al., 2018;Vitasse et al., 2019) achieve climatic reconstructions and ecological investigations (Huang et al., 2017) through retrospective measurements of ring width or, more recently, basal area increments (Biondi & Qeadan, 2008). Typically, dendroecology estimates and employs a response curve that corresponds to a RN calculated at the level of the group of individuals (population): it thus estimates the average plasticity of this group of individuals, and it can be called average plastic response (Feinard-Duranceau et al., 2018). Other physical and anatomical ring variables are rarely used as alternatives to ring width (Fonti & Jansen, 2012). Non-significant average plastic response may conceal very variable levels of PP, which can be elucidated only by fitting RN at the individual level, and fails to achieve the key-question about the variability and significant differences between entities (groups of trees, populations, genetic entities) for PP (Arnold et al., 2019). But developing significant 1 RN, that is, significant phenotype-environment relationships at the genotype, that is, at the individual level, is challenging and rarely achieved in PP studies (Arnold et al., 2019). This may explain why, while the amount of dendroecology studies estimating an average plastic response is very high, and while, to our knowledge, the first published study of inter-ring PP is relatively old (Fallour-Rubio et al., 2009), the number of published studies estimating PP by means of individual inter-ring RN is still very low today.
The retrospective use of tree-rings for analysis of PP with construction of RN involves two different time-scale approaches: interand intra-ring PP. The inter-ring approach considers inter-annual PP, while the intra-ring methodology investigates intra-annual (or intragrowing season) PP. Studies of inter-ring PP associate inter-annual time-series of ring variables to inter-annual time-series of climatic variables, like for example ring width and mean summer temperature (Fallour-Rubio et al., 2009) or ring width and a climatic drought index (Marchal et al., 2019). Intra-ring PP considers the construction of a RN associating intra-ring time-series with intra-annual environmental variables, such as ring microdensity profiles and water balance variation during the year (Sánchez-Vargas et al., 2007). Intra-annual ring fluctuations (IADF) are also used as a way to quantify intra-ring PP (Balzano et al., 2019;Nabais et al., 2014).
The shape of the RN, linear or non-linear, determines the variables used for eventually estimating the PP. If the RN is linear, the slope of the straight regression line is the unique PP variable (Arnold et al., 2019;Sánchez-Vargas et al., 2007). If the RN is non-linear, then several variables may be necessary to correctly quantify the PP (Martinez-Meier et al., 2009). These authors proposed the word "dendroplasticity" to describe the retrospective estimation of PP based on tree-ring analysis.
In this study, we use microdensity tree-ring variables of European Larch (Larix decidua Mill.) to estimate inter-annual PP to temperature, aiming at maximizing the number of trees with significant RN estimation at all elevation levels and for all ring variables. Then we study PP variation between individuals and between contrasted environments, that is, four study plots along an elevational gradient in a naturally regenerated adult Larch forest. We selected microdensity variables considering that density is a proxy for important xylem ecological (Björklund et al., 2019) and functional properties (e.g. hydraulic efficiency and safety and mechanical stability) (Lachenbruch & McCulloh, 2014).
European larch is a key forest tree species all over the Alps and in certain regions of Central Europe. In the southwestern part of its natural area, Larch is distributed along elevation gradients starting as low as 1,200 m and culminating as high as 2,500 m. It is one of the mountain species with the major temperature variation along its elevational gradients (Carrer et al., 1998;Fourchy, 1952;Jochner et al., 2017;Obojes et al., 2018;Saulnier et al., 2019). Elevational gradients are the most powerful natural experiments to test ecological responses to geophysical influences, such as temperature (Körner, 2007). Individuals in colder conditions increase their growth when temperature increases, while on the contrary individuals in warmer condition reduce their growth when temperature increases, with a smaller between-individual variation at the warmer site (Clark et al., 2003;Way & Oren, 2010). This suggests that their response to time-related temperature variation and specifically to global warming is distinct at the different temperature conditions of the various elevational levels (Morin et al., 2018). Populations distributed over an environmental gradient experience differential selection pressures (Barton, 1999), which can ultimately lead to different levels of intrapopulation variation and inter-population genetic differentiation for adaptive traits. Local adaptation occurs when populations that experience divergent selection become better adapted to their own local environment than other populations, and comparatively less adapted to neighboring ones (Kawecki & Ebert, 2004). The other possibility is that the larch forest distributed along this gradient be one predominantly homogeneous population with considerable gene flow between all elevational levels . In fact this pattern was observed with the same trees used in this study along the same elevational gradient for neutral genetic markers (Nardin et al., 2015).
Whether variation of larch PP reflects local adaptation or neutral genetic variation is among the questions raised by this study.
Our objective was to fit inter annual-ring reaction norms of several ring variables and use their variations in sign, magnitude and range to estimate PP variation among elevational plots and ring variables. Finally, we discuss whether elevational variation for PP could correspond to local adaptation and could improve larch response to climate change.

| Sites characteristics
Our study site is located in the native range of Larix decidua (Mill.) at Villard-St-Pancrace, close to Briançon (latitude: 44.9° N; longitude: 6.65° E; average elevation: 1,326 m) in the French Alps. The experiment is formed of four forest plots distributed along a north-faced Alpine steep-slope ranging from 1,200 m to 2,500 m and mostly covered by uneven-aged larch forests with some patches of Abies alba, Pinus sylvestris, Pinus cembra and Pinus uncinata. The four plots each include 200 larch trees and are located at 2,300, 2,000, 1,700 and 1,350 m a.s.l., respectively (Nardin, 2013;Nardin et al., 2015). The average annual maximum temperature difference along the gradient is 6.1°C, changing from 7.58°C at the highest site to 13.68°C at the lowest one (Table 1; Table S1). Besides temperature, other factors such as solar radiation (higher at the lowest plots), and soil water availability and fertility (higher for the two intermediate plots) are changing. These differing conditions affect the tree characteristics at each plot with large variation on average tree height (from 16 to 27 m), stem girth (from 81 to 112 cm) and average annual radial growth (from 0.63 to 0.91 mm; Table 1; Table S1).

| X-ray, cross-dating and detrending
All the details about the methods of tree-ring related measurements are in (Nardin, 2013;Nardin et al., 2015;Rozenberg, Chauvin, et al., 2020;. Shortly, from each selected tree, we collected a 5.5 mm diameter-increment core at 1.3 m in the stem using a powerdriven Pressler increment borer. The increment cores were stored in polycarbonate honeycomb boxes and dried to reach a stable uniform humidity of about 12%. The cores were sawn to a uniform crosssection thickness of 2 mm and immersed during one week in pentane solvent (C 5 H 12 ) to extract resin. The samples were dried again, X-rayed and the X-ray films were scanned at 4,000 dpi, then analyzed with WINDENDRO (Windendro 2008e Regent instruments, Canada). We cross-dated the tree-ring time-series using the Interdat software (version 1.1, Becker et al., 1994;Mérian et al., 2011) and pointer years based on the tree-ring width (RW) measurements provided by WINDENDRO. We considered the rings 1967-2007 (41 successive years). We used the "extreme average method" (Vargas-Hernandez & Adams, 1991) to separate each annual ring microdensity profile into its earlywood and latewood components. We then obtained the widths (RW), the average densities (RD) of each dated annual ring, specifically the earlywood, latewood width (EW and LW), and the earlywood and latewood average density (EWD, LWD).
We used and compared several methods to correct the timeseries of ring microdensity variables to only keep climatic variability (Rozenberg, Chauvin, et al., 2020). Our objective was to estimate individual RN on the maximum number of trees. However, fitting RN with statistically significant slopes is often a big challenge in PP studies. Here we compared different methods used at the successive steps of the RN construction, for their ability to maximize the number of trees with significant RN. First we tested several detrending methods: we used no correction, negative exponential curve (Cook & Kairiukstis, 1990;Fritts, 1976) and regional curve standardization (RCS; Esper et al. 2003), with residual RCS and ratio RCS (Esper et al., 2003;Rozenberg, Chauvin, et al., 2020). For the negative exponential function we used the dendrochronology library in R, dplR (Bunn, 2008). Since larch budmoth (LBM) can induce major cyclic defoliations with important non-climatic impact on ring microdensity profiles (Castagneri et al., 2020;Peters et al., 2020; we also tested the impact of standardizing for

| Construction of the reaction norms and estimation of phenotypic plasticity
To fit the reaction norm (RN) between the tree-ring variables and the climate variables we applied the linear model Y i = β1 + β2X i + ϵ ij using the lm() function of R (R Core Team, 2018), where Y i is the phenotypic variable; X i is the climatic variable, β1 is the intercept and β2 is the slope (the PP value), and ϵ ij is the error term (the residuals).
The adjusted model R-square is used to estimate the quality of the   we used the Briançon data only. For each plot we adjusted the Briançon climatic time-series from the elevation effect, as in (Latreille et al., 2017;Rozenberg, Chauvin, et al., 2020) using the on-site climatic data measured since 2008 at each plot.
We estimated the RN by averaging the daily climatic data for time-windows of variable duration and position during the calendar year. We compared four methods (M1 to M4). In M1 we fitted the RN with the same annual climatic means for all the trees in the four plots. In M2 we estimated the RN for a time-window based on the results of a ring formation study (Saderi et al., 2019). It reflects the duration of the ring formation period estimated on a sub-sample of the same trees, studied during the 2013 growing season (Saderi et al., 2019). In M3 we systematically tested different time-windows and at each elevation level. We selected one common time-window for all the trees in each plot: this time-window is the one giving the highest number of trees with significant RN. Finally in M4, we tested the time-windows at the tree level: for each tree we selected the time-window giving the RN with the highest R-square. We obtained similar results for minimum and maximum temperature. M3 and M4 gave a higher proportion of trees with significant RN than M1 and M2. In contrast we were not able to fit RN with statistically significant slopes (p < .05) with precipitation, whatever the method, which was then excluded from the study. Figure S2 in Supporting Information shows the heatmap of the tested flexible time-windows in M4 for a given tree, for LWD and maximum temperature.
The slopes are used as quantitative estimates of the tree PP. We used a single factor fixed effect ANOVA to test for the differences between the elevational plots.

| Diversity of the RN along the gradient
The main characteristics of the trees in the four elevational plots (table 2) showed that on average the trees were slightly bigger at 1,700 m, taller at 1,700 and 2,000 m, younger at 2,300 m and with a lower wood density at 1,350 m.

| Test and selection of the methods (standardization and construction of RN)
We observed that the standardization method to adjust from the cambial age effect did not affect the results: the number of trees with significant RN was not strongly modified and the results were the same with the different standardization methods tested. We also found that the larch budmoth standardization did not affect the results since it did not change the general findings (data not shown).
In contrast, the number of trees with significant RN varied a lot between the four fitting methods M1 to M4. M3 and M4 gave the highest number of trees with significant RN. For most ring variables and at most elevations, the number of trees with significant RN was higher for M4 than for M3, with a difference ranging from six to 46 trees, corresponding to four to 30% of the total number of trees in the plot ( Figure 1). The superiority of M4 over M3 was stronger for RW at 1,350 and 1,700 m and for EWD and LWD at 1,700 m ( Figure 1). The only exception was LWD at 2,300 m, where there were 115 trees with significant RN with M3 and 114 for M4 ( Figure 1).
We also compared the value of the slopes of the RN fitted with M3 and M4. We found that they were very strongly correlated, with r ranging from 0.94 to 0.98 according to the ring variable ( Figure 2).
Finally, we observed that, when significant, the main trends of the findings were similar between M3 and M4.  only four site-specific slopes resulted to be higly significant (p < .01), namely RW and LWD at 2,000 and 2,300 m a.s.l. (Figure 3; Figure S3 in Supporting Information) whereby the two at 2,300 were stronger than at 2,000 m. The other site average RN slopes were not significant, except for RD for which we found a significant and low relationship at 2,000 m a.s.l. (r = 0.38, p =.014, not shown).  Table S2 in Supporting Information).
Depending on elevation, the percentage of significant slopes ranged between 53% and 66% for EWD, 73 and 94% for LWD, 63 and 71% for RD and 75 and 94% for RW (Figure 4). For EWD and RD at all

| Time-windows
The individual time-windows used to estimate the significant RN in M4 are presented in the Supplementary Material (Table S3;

| Variation of the slope value and of the PP intensity
Inside each group of slopes, positive and negative, there was variation of the value of the slope, that is, variation of the intensity of PP.
For most variables and types of PP this intensity of PP is significant between elevational levels ( Figure 5). In two cases only (EWD for positive PP and RW for negative PP) the elevational variation was not significant. In one case (RD for positive PP) the variation was significant at the 5% probability level. The corresponding trend was a consequence of steeper slopes at 1,350 m (Table 3). But the corresponding number of trees was only 10, while it was respectively 38, 72 and 69 at the other elevational levels, between which there was no significant variation of PP ( Figure 5; Table 3). For the five other cases, the variation of the PP values was significant at the 0.1% probability level. In these cases, the highest PP values (positive and negative) were always found at 2,000 or 2,300 m a.s.l., while the lowest were always found at 1,350 or 1,700 m a.s.l. However, in some cases, these minimum and maximum PP values were estimated with a low or very low number of trees: for RD, the mean value of the negative PP at 2,000 m a.s.l. was estimated with 12 trees. Consequently, there was no significant difference between the three higher elevations (1,700, 2,000 and 2,300 m a.s.l., Table 3). For LWD at 2,000 and 2,300 m a.s.l., the mean value of the negative PP was estimated with respectively eight and one trees only. Here again, there was no significant difference between the three higher elevations. The significant elevational variation of the PP value was better described for LWD (positive and negative PP), RW (positive PP) and for EWD (negative PP) (Figure 4). The details about the significance of the variation between pairs of elevational levels in Table 3 show that for LWD positive PP, the value at 1,700 m was strongly and significantly lower than that of the PP at the higher elevational levels. For RW positive PP, the elevational trend was highly significant between each pair of successive elevational levels. Finally, for EWD, only the value of the negative PP at 1,350 m asl was significantly different from that at the three higher elevations (Table 3). We found individuals with slopes of reaction norms in opposite directions for different plasticities, that is, different associations of phenotypic and environmental variables. This is common and reflects the specific physiological process associated with each phenotypic variable (Pélabon et al., 2013). For each of the four ring variables studied, we found a broad range of individual tree plasticities within a natural population: for a given trait, they range from positive to negative slope values. This population includes as well a more or less large proportion of individuals (from 6% to 41%) with no PP related to the climatic variable studied (null plasticity).

| D ISCUSS I ON
There are other examples of slopes of reaction norms in opposite directions for a given PP in animals (fishes and insects) and in plants, but we found none in trees. The variation between positive and negative slopes was sometimes associated with age (Smoliński et al., 2020), more often with quality of environment (Diamond & Kingsolver, 2012;Tammaru & Teder, 2012;Teder et al., 2014), and in the case of field gentian (Gentianella campestris), with population variation (Juenger et al., 2000). In our case, the variation between positive and negative slopes slopes of the RN is related with elevation for three of the four ring variables. Across all variables, the slopes have a tendency to be more negative at the low elevation and more positive at the top. Furthermore, the individuals with a negative slope at the low elevation plots (1,350 and 1,700 m) tend to have a more negative slope than the individuals from the high elevation plots (2,000 and 2,300 m). The more positive slopes at the higher elevation plots for the trees with a positive slope were found mainly for RW and LWD.
Our results mean that a temperature increase is indeed associated to opposite reactions between trees within a population: the temperature increase corresponds, according to individual trees, to either an increase or decrease of ring width, ring density, earlywood

| Strength and weaknesses of the flexible timewindows
As stated immediately above, we carefully compared the results of the analysis conducted with fixed and flexible time windows. We observed that the number of trees with significant reaction norms was lower with the fixed time windows method. The flexible time window method better took into account the differential behaviour of the individuals towards temperature. If at species or even population levels, mean annual (or growing season) temperature is convenient and satisfactory to explain overall radial tree growth, it may not be the case anymore when the target is the individual response. One consequence was that, especially for earlywood density, the statistical power of between-elevation-level comparison decreased for the fixed-window method in comparison with the flexible-window method: some of the differences between the elevation levels were not statistically significant. We also screened the strength of the relationships between the ring variables and temperature for each individual using correlation heat-maps for the flexible time-windows (example tree 41 for LWD, Figure S2 in Supplementary Material).
The results showed that for most trees, the range of the first and last days of the time-windows displaying significant correlation was usually large and covered a wide period. For example ( Figure S2 in Supplementary Material), a change in the correlation coefficient value from 5% to 10% is observed within a 2 to 3 months time-window change.
It means that in the tested range, the choice of the timewindow did not affect much the value of the correlation coefficient, because it did not affect much the interannual variation of the average temperature of the time-window. Conversely, the temperaturesensitive periods were rather variable between the ring variables and the elevational plots ( Figures S4-S7, Supplementary Material).
As expected, days that maximize the relationship with ring growth and density parameters are delayed at higher elevations compared to lower elevations. If we assume that there is a close relationship between these time-windows and the phenology of ring formation, we would expect for example earlier periods for earlywood than for latewood (Rossi et al., 2013;Saderi et al., 2019) and shorter periods for the high elevation plots (He et al., 2012;King et al., 2013). However the variation observed among plots and traits is not fully consistent with what could be expected from a biological point of view. The time-windows we defined are not supposed to precisely determine starting and ending days of growth processes but are rather climatic periods maximizing statistical relationships without any attempt to physiologically interpret them. These annual periods are convenient tools to maximize the number of trees with significant estimations of the RN and to increase the statistical power of the experiment, but should not be strictly interpreted from a biological point of view.

| Variation of phenotypic plasticity (PP) along the elevation gradient
We used the significant (positive or negative) slopes of the RN as quantitative estimates of PP. For the trees with non-significant RN, we cannot conclude if the non-significant estimation of the slope corresponds to a flat reaction norm (Fusco & Minelli, 2010), or null PP, or to a failure to estimate the PP (the individual is plastic but we were not able to estimate its PP with the available data). For convenience, we call trees with "null PP" the trees for which the estimation of the slope was not statistically significant (not significantly different from zero). For an adaptive trait, the distinction between positive and negative slopes could correspond to a major (genetic?) divergence between responses. For latewood density, and to a lesser extent, for ring width and ring density, the change from negative to positive slopes is closely associated with elevational variation. The proportion of trees with a positive PP tends to increase from the bottom to the top of the gradient, with obviously the opposite trend for the negative PP. This tendency is especially strong for LWD (latewood density): in the warmer conditions at the bottom of the gradient, the temperature increase is associated to a latewood density decrease for most of the trees, while in the colder conditions at the top of the gradient, a temperature increase is associated to a latewood density increase for nearly all the trees.
This trend is less marked for RD (mean ring density), which logically Information at this level is however key to study the PP potential of evolutionary adaptation, which relies on phenotypic and genetic variation, and on heritability.
The linear warming trend observed at Briançon explains about half of inter-annual temperature variation between 1967 and 2007 ( Figure S1 in Supporting Information). In another study, a similar trend was found for the 1967-2016 period at the same location (Rozenberg, Chauvin, et al., 2020). If we assume that this linear temperature increase is a manifestation of the global warming, then about half of the PP is an individual response to this global warming.
According to (Rozenberg, Chauvin, et al., 2020), this global warming effect is stronger for tree response at low elevation than at high elevation: global warming reveals the potential of plasticity of larch.
Whether such response can be considered as an adaptation mechanism relies on the adaptive value, or fitness, of PP. To what extent the fitness of PP is related to the fitness of the phenotypic trait (Nicotra & Davidson, 2010;Via et al., 1995)? Obviously, and as observable for example in Figure 3, the range of temperature variation at each elevation is shorter than the complete range of interannual temperature variation along the whole gradient.
What would be the shape of a reaction norm of a tree facing this For some ring variables and/or at some elevation, there is a mixture of trees with positive and negative slopes. Such opposite PP could correspond to differences between trees for the position of the complete nonlinear reaction norm along the temperature axis, as shown in Figure 7.
This variation could reflect both environmental and genetic differences between the trees. The proportion of between-tree environmental and genetic variation could be very different for trees sharing the same plot or being largely separated along the gradient. However, test of these hypotheses requires access to genetic information, which necessitates other approaches such as genetic trials. The experimental trial in this study does not encompass a common garden experiment and thus does not permit the accurate separation of the environmental and genetic components of PP variation. A previous study of genetic diversity along the same elevational gradient (Nardin et al., 2015) showed a genetic structure only slightly affected by climatic variation, human activities or historical events. Yet, a small but significant inter-plot genetic variation indicates the existence of variable genetic dynamics, which could be an indication of local adaptation (Nardin et al., 2015). This suggests that the strong gene flow between all elevations levels redistributes more or less the same large genetic diversity everywhere at each new generation. This large genetic diversity could be related with the large PP variation observed along the gradient and especially at mid-gradient. Indeed, large phenotypic variation is often associated to large genetic variation (Roff, 1995). In addition, the more constraining climatic conditions at the gradient extremities could locally select adapted trees and reduce phenotypic and genetic variation, including for PP. But these facts are not enough to provide evidence for genetic variation, local adaptation and a potential for evolutionary adaptation for PP in this gradient.
Studies based on genetic trials, reciprocal transplant experiments and non-neutral genetic markers would deliver additional relevant information.

| Strength of the individual reaction norm approach
A PP variable obtained from the reaction norm can be seen as a Whereas this study shows that PP tends to be greater in the colder conditions at the top of the gradient, and more variable in the most favourable conditions at mid gradient, there are also very strong differences between ring variables: it does not seem possible to consider PP as a single trait. PP should be systematically referred to mentioning the corresponding phenotypic trait-environmental variable association. Estimating reaction norms at the individual tree level provides information about the inter individual variation of PP, which is not available when it is studied as the average plastic response of a group (species, population) of trees. This information is especially relevant in evolutionary studies when the adaptation potential of the population submitted to new selection pressure such as climate change is heavily dependent of the available genetic diversity of its individual tree components.

CO N FLI C T O F I NTE R E S T
The authors declare no conflict of interest.
[Correction added on 18 June 2021, after first online publication: Conflict of Interest statement added to provide full transparency.]

ACK N OWLED G M ENTS
The