\documentclass[10pt]{article}

\usepackage{fullpage}
\usepackage{setspace}
\usepackage{parskip}
\usepackage{titlesec}
\usepackage[section]{placeins}
\usepackage{xcolor}
\usepackage{breakcites}
\usepackage{lineno}
\usepackage{hyphenat}





\PassOptionsToPackage{hyphens}{url}
\usepackage[colorlinks = true,
            linkcolor = blue,
            urlcolor  = blue,
            citecolor = blue,
            anchorcolor = blue]{hyperref}
\usepackage{etoolbox}
\makeatletter
\patchcmd\@combinedblfloats{\box\@outputbox}{\unvbox\@outputbox}{}{%
  \errmessage{\noexpand\@combinedblfloats could not be patched}%
}%
\makeatother


\usepackage{natbib}




\renewenvironment{abstract}
  {{\bfseries\noindent{\abstractname}\par\nobreak}\footnotesize}
  {\bigskip}

\titlespacing{\section}{0pt}{*3}{*1}
\titlespacing{\subsection}{0pt}{*2}{*0.5}
\titlespacing{\subsubsection}{0pt}{*1.5}{0pt}


\usepackage{authblk}


\usepackage{graphicx}
\usepackage[space]{grffile}
\usepackage{latexsym}
\usepackage{textcomp}
\usepackage{longtable}
\usepackage{tabulary}
\usepackage{booktabs,array,multirow}
\usepackage{amsfonts,amsmath,amssymb}
\providecommand\citet{\cite}
\providecommand\citep{\cite}
\providecommand\citealt{\cite}
% You can conditionalize code for latexml or normal latex using this.
\newif\iflatexml\latexmlfalse
\providecommand{\tightlist}{\setlength{\itemsep}{0pt}\setlength{\parskip}{0pt}}%

\AtBeginDocument{\DeclareGraphicsExtensions{.pdf,.PDF,.eps,.EPS,.png,.PNG,.tif,.TIF,.jpg,.JPG,.jpeg,.JPEG}}

\usepackage[utf8]{inputenc}
\usepackage[T2A]{fontenc}
\usepackage[ngerman,greek,polish,english]{babel}



\usepackage{float}








\begin{document}

\title{Friends or foes? Plant-animal coevolutionary history is driven by both
mutualistic and antagonistic interactions}



\author[1]{Lisieux Fuzessy}%
\author[2]{Juan Antonio Balbuena}%
\author[3]{Omer Nevo}%
\author[4]{Jadelys Tonos}%
\author[5]{Bastien Papinot}%
\author[6]{Daniel Park}%
\author[7]{Daniel Sol}%
\author[8]{Kim Valenta}%
\author[4]{Onja Razafindratsima}%
\author[9]{Miguel Verdu}%
\affil[1]{UNESP Campus de Rio Claro}%
\affil[2]{Universitat de València}%
\affil[3]{German Centre for Integrative Biodiversity Research (iDiv) Halle-Jena-Leipzig}%
\affil[4]{University of California Berkeley}%
\affil[5]{University of Iceland}%
\affil[6]{Purdue University}%
\affil[7]{CREAF}%
\affil[8]{University of Florida}%
\affil[9]{CSIC}%


\vspace{-1em}



  
  \date{May 16, 2024}


\begingroup
\let\center\flushleft
\let\endcenter\endflushleft
\maketitle
\endgroup





\selectlanguage{english}
\begin{abstract}
Coevolution played a central role in shaping biodiversity. However,
coevolutionary events driving reciprocal diversification between
interacting partners lack empirical evidences. Examples of
diversification arising from mutualisms and antagonisms at different
trophic levels are scarce, which limits our understanding on how complex
relationships between species arise within communities. By adopting a
cophylogenetic framework, we investigated whether congruence in
plant-lemur phylogenies are driven by mutualisms and antagonisms in
Madagascar, where endemic species have evolved within a unique isolated
biogeographical context. Although we found weak support for coevolution,
this is not to say that interactions between lemurs and plants did not
promote a shared evolutionary history. Weak cophylogenetic signals do
not necessarily imply lack of co-diversification. Rather, our results
suggest that vertebrates and plants influenced each other evolution
through ecological interactions, but in a multi-specific and generalist
context where the cophylogenetic process leaded to spatio-temporal
asymmetries and shifts between periods of coevolution and independent
evolution, ultimately resulting in a weak, continuous and diffuse
process.%
\end{abstract}%



\sloppy


Coevolution played a central role in shaping biodiversity. However,
coevolutionary events driving reciprocal diversification between
interacting partners lack empirical evidences. Examples of
diversification arising from mutualisms and antagonisms at different
trophic levels are scarce, which limits our understanding on how complex
relationships between species arise within communities. By adopting a
cophylogenetic framework, we investigated whether congruence in
plant-lemur phylogenies are driven by mutualisms and antagonisms in
Madagascar, where endemic species have evolved within a unique isolated
biogeographical context. Although we found weak support for coevolution,
this is not to say that lemurs and plants did not share evolutionary
history. Weak cophylogenetic signals do not necessarily imply lack of
co-diversification. Rather, our results suggest that vertebrates and
plants influenced each other evolution, but in a multi-specific context
where the cophylogenetic process leaded to spatio-temporal asymmetries
and shifts between periods of coevolution and independent evolution,
ultimately resulting in a weak, continuous and diffuse process.

\textbf{INTRODUCTION}

Coevolution ---the reciprocal adaptation between two or more interacting
organisms (Hembry et al. 2014)--- is considered to have played a central
role in shaping the diversity of life on Earth (Laine 2009). Yet,
empirical evidence that coevolution is the main mechanism driving
reciprocal diversification among interacting clades lack alternative
explanations (Medina et al. 2022). Most examples of diversification
driven by coevolution involves cases of resource competition between
species during adaptive radiations (reviewed in Hembry et al. 2014). In
contrast, examples of diversification arising from mutualistic and
antagonistic interactions at different trophic levels are comparatively
rare (e.g. Hutchinson et al. 2017; Fuzessy et al. 2022). The lack of
attention to such pervasive and relevant types of interactions not only
leads to a biased view of the importance of coevolution in
diversification processes, but also limits our understanding on how
complex relationships between species arise within communities (Thompson
1994).

A major obstacle to generate empirical evidence for coevolutionary
history in antagonistic/mutualistic interactions has been ruling out
alternative explanations of diversification (Althoff et al. 2014; Medina
et al. 2022). Although often taken as evidence for coevolution,
reciprocal adaptation can emerge through adaptation by one of the
interacting species alone or through neutral processes, such as genetic
drift (Yoder \& Nuismer 2010). Plant-animal interactions are recognized
as important drivers of one-sided evolution (Strauss \& Irwin 2004;
Hembry \& Weber 2020; Johnson et al. 2021). The correlation in features
of interacting pairs, such as floral structures that match traits of
pollinators (Nilsson 1988), color vision phenotypes that shape fruit
color (Valenta et al. 2018), or defenses against predators that are met
by counter defenses (Brodie \& Ridenhour 2003), may also arise through
correlated selection (Nuismer et al. 2010). Phylogenetic congruence
between the interacting lineages, another major expectation of
coevolutionary processes, is neither exempt of problems. While often
interpreted as evidence of co-speciation, in nature, other types of
evolutionary events often act concurrently, so a perfect cophylogenetic
fit is rarely observed (Hutchinson et al. 2017; Fuzessy et al. 2022).
Congruence may result from a combination of events unrelated to
co-speciation, such as phylogenetic tracking (when one taxon tracks the
evolution of its interaction partner), shared biogeographic histories
(like vicariance, where common biogeographic events cause the
subdivision of the distribution range of both interacting partners) or
from parallel evolution driven by climate changes and other broad
selective agents (Nuismer et al. 2010).

Demonstrating that interactions shape concomitant diversification on
both sides has also been challenging due to two main constraints. First,
interacting species on both sides are on a continuum of
generalist-specialist interactions, which strongly affects the expected
selective pressure each party exerts on the other, and hence the overall
selective regime. In plant-animal interactions, phylogenetic tracking by
adaptive switching behaviors, the ability of generalists to adaptively
increase the consumption of one resource at the expense of decreased
consumption of an alternative resource (Carnicer et al. 2008), is
expected to be particularly relevant in creating phylogenetic
congruences that can be confounded as coevolutionary diversification.
This is because switching behaviors should not only facilitate animals
to change feeding habits to new plants at the expense of others, but
morphology and other constraints should also increase the chances that
changes will favor plants that are closely related (and thus more
similar) to those previously used as a resource (see Carnicer et al.
2008). The second constrain is that mutualism and antagonism themselves
are only two ends of a single continuum, where no individual acts
exclusively as an antagonist or a mutualist. For example, animals that
feed on flowers can carry a significant amount of pollen in their body,
acting simultaneously as florivores and pollinators (Kress et al. 1994).
In the same vein, when feeding from fruits, many frugivores transport
the seeds far from parent trees, acting as seed dispersers (Schupp et
al. 2010). However, since not all seeds are defecated intact, seed
dispersing animals can also act as seed predators (Norconk 2020). Due to
these constrains and the difficulties to rule out alternative
explanations, the role of coevolutionary processes in
antagonistic/mutualistic interactions is currently backed by limited
evidence (but see Hutchinson et al. 2017; Fuzessy et al. 2022).

While generating macroevolutionary evidence for coevolutionary
diversification in antagonistic/mutualistic interactions remains
challenging, recent advances in analytical tools developed for testing
phylogenetic congruence provides a stronger framework to rule out
alternative explanations of diversification (Balbuena et al. 2013;
Blasco-Costa et al. 2021). A great advantage of these cophylogenetic
methods (so called pattern-based methods, Dismukes et al. 2022) is the
ability to use phylogenies where interacting species may have multiple
pairs, which is the case of most plant-animal interactions. This allows
to accommodate variation among the interacting species along the
generalist-specialist continuum while considering phylogenetic
uncertainties. With this framework, it is also possible to evaluate the
contribution of individual links between interacting partners to the
overall congruence (Hutchinson et al. 2017), and to test the extent to
which the patterns of co-diversification and/or coevolution can be
explained by phenotypic traits (Blasco-Costa et al. 2021).

Here, we adopt the above cophylogenetic framework to investigate
macro-coevolutionary hypotheses for phylogenetic congruence of
plant-lemur interactions in Madagascar. The flora and fauna of
Madagascar provides an interesting study case to test such
macro-coevolutionary hypotheses. About 82\% of vascular plants
(Callmander et al. 2011) and 84\% of vertebrates (Goodman \& Benstead
2005) are endemic, and hence have evolved under similar climatic
conditions within a relatively small island, and have had ample
opportunity for interaction and coevolution (G\selectlanguage{ngerman}énin et al. 2022). Lemurs,
in particular, diverged from Haplorrihini primates about 68 Ma, having
originated from an ancestor on the African mainland, and occurring only
in Madagascar (Pozzi et al. 2014). They arrived in the island earlier
than other mammalian orders, resulting in a variety of adaptations to
diverse niches and feeding habits, which include the consumption of
fruits, leaves, flowers, nectar, bark and insects (Richard \& Dewar
1991; Dewar \& Richard 2012).

Previous work suggests that lemur colonization and subsequent
diversification may have been facilitated by the divergence of endemic
Malagasy angiosperms. Based on physiognomic and chronological
reconstructions, Génin et al. (2022) concluded that the generalization
of mutualistic interactions (such as frugivory and nectarivory) might
have boosted the coevolution of modern lemurs and at least 10 new
angiosperm families. However, observations were based on physiognomic
and chronological reconstructions, and empirical evidence remains
elusive both for pattern and process. Also, we still lack clear evidence
for the potential of mutualistic and antagonistic interactions in
generating congruence in the phylogenies of lemurs and Malagasy
angiosperms. Here we address these issues by testing for cophylogenetic
signals across a spectrum of plant-lemur interactions, including
frugivory, folivory, florivory, granivory and nectarivory. To support a
role for coevolutionary processes, we first expect phylogenies of lemurs
and plants to be congruent. This is to be expected if mutualistic and
antagonistic interactions shape reciprocal evolution regardless of
interaction type. We also expect to identify putative coevolving
phenotypic traits that may have propelled diversification (Blasco-Costa
et al. 2021). If both predictions met, we finally expect to find a
positive correlation between phylogenetic distances and dissimilarities
in sets of interacting partners for both lemurs and plants. This would
help to rule out the possibility that the observed cophylogenetic
patterns were driven by phylogenetic tracking or vicariance instead of
coevolution (Kahnt et al. 2019).

\textbf{METHODS}

\textbf{Database}

\emph{Plant-lemur interactions}

We used comprehensive information on plant-lemur interactions from the
Lemur Food Plants database (LFP, Steffens 2020), updated with new
information published between October 2019 and August 2022 (SM Table
S1.1). The final dataset comprised 6,061 pairwise interactions (SM Table
S1.2), where same species were recorded interacting with distinct types
of resources (fruits, flowers, nectar, leaves). This is reflected in the
dataset, which included 3,317 frugivory interactions (1,519 unique),
2,249 folivory interactions (1,056 unique), 717 florivory interactions
(367 unique), 207 nectarivory interactions (50 unique) and 131 granivory
interactions (58 unique). Rarefaction curves of sampling effort are
presented in SM Figure S5.

\emph{Plant and lemur traits}

To test our coevolutionary hypothesis, we used a functional trait
approach. For plants, we obtained information on traits related to
fruit-consumption (fruit length and width, seed length and width, number
of seeds per fruit and diaspore color) from Albert-Daviaud et al. (2021)
(SM Table S2). For lemurs, we obtained information on morphological
traits known to be related to food consumption (body mass, body length,
and tail length) (Mittermeier et al. 2010; SM Table S3).

We also gathered complementary information on lemur activity patterns
and feeding guilds, which are related to feeding behavior (Mittermeier
et al. 2010; Razafindratsima et al. 2018; Galán-Acedo et al. 2019; SM
Table S3), and on plant traits that may be related to selection by
lemurs according to interaction types: frugivory, folivory and
florivory. Full description of plant traits is available in Table 1 and
the complete list of traits is available in the Table S4. The
complementary information on fruit, leaf and flower traits were
available for a restricted number of plant species for which we had
information on lemur-plant interactions. Thus, this complementary
information was used only to test for their possible influence on the
contribution to the cophylogenetic signal according to each type of
interaction they are related (see `Individual contributions to the CS'
section below).

\textbf{Table 1 -} Full description of plant traits\selectlanguage{english}
\begin{longtable}[]{@{}llllll@{}}
\toprule
\textbf{Interaction type} & \textbf{Plant trait} & \textbf{Measure} &
\textbf{Description} & \textbf{\# Plant spp. (total)*} &
\textbf{Source}\tabularnewline
\midrule
\endhead
Frugivory & Fruit size & Length, width & Length and width measures & 696
(696) & Albert-Daviaud et al. (2021)\tabularnewline
& Seed size & Length, width & Length and width measures & 696 (696)
&\tabularnewline
& Seeds/fruit & \# seeds/fruit & Count of number of seeds per fruit &
696 (696) &\tabularnewline
& Fruit scent & Quantitative & & 18 (696) & Nevo et al.
(2018)\tabularnewline
& & Chemical & & &\tabularnewline
& Fruit color & Subjective color & Human-defined subjective color
categories (contrast, black, blue, brown, red, orange, yellow, green,
blue, purple) & 404 (696) & Albert-Daviaud et al. (2021)\tabularnewline
& & Hue & Hue subdivisions converted into a continuous variable relative
to color wavelengths. Pure red corresponds to zero and shorter
wavelengths, while blue-purple corresponds to increasing values & 57
(696) & Razafindratsima \& Dunham (2019)\tabularnewline
& & Chromatic distances & Chromatic contrasts between fruits and
background leaves under a diurnal, forest shade illuminant in two visual
channels: red--green and yellow--blue\textsuperscript{1} & 18 (696) &
Nevo et al. (2018)\tabularnewline
& Fruit type & Fruit type & Classification based on morphology of the
gynoecium and pericarpium & 396 (696) & Albert-Daviaud et al.
(2021)\tabularnewline
& Nutritional content & Nitrogen & First and second axis of a
PCA\textsuperscript{2} (content of N, soluble proteins, sugar and lipid
per dry mass of fruit pulp, in mg/g) & 42 (696) & Razafindratsima \&
Dunham (2019)\tabularnewline
& & Protein & & &\tabularnewline
& & Sugar & & &\tabularnewline
& & Fat & & &\tabularnewline
& Dispersal mode & Lemur/mixed & Whether if lemur-dispersed or dispersed
by lemurs and other frugivores (mainly birds) & 86 (696)
&\tabularnewline
Folivory & Leaf form & Leaf area & Area of the smallest lamina unit
(leaf or leaflet, in mm\textsuperscript{2}) & 93 (673) & D\selectlanguage{ngerman}íaz et al.
2022\tabularnewline
& & Leaf thickness & & 72 (673) & TRY database (Kattge et al.
2020)\tabularnewline
& Leaf mass & LMA & Leaf mass per leaf area (in g/mm2) & 70 (673) & D\selectlanguage{ngerman}íaz
et al. 2022\tabularnewline
& Water content & Water & Water content per leaf dry mass (in ml/g) & 22
(673) & TRY database (Kattge et al. 2020)\tabularnewline
& Nutritional content & Nitrogen & N content per dry mass (in mg/g) & 66
(673) & TRY database (Kattge et al. 2020)\tabularnewline
& & Phosphorous & P content per dry mass (in mg/g) & 51 (673)
&\tabularnewline
Florivory & Flower color & Subjective color & Human-defined subjective
color categories (white, pink, red, orange, yellow, brown or
blue-purple) & 49 (267) & TRY database (Kattge et al.
2020)\tabularnewline
& Flower shape & Symmetry & Whether flower perianth can be divided into
two or more identical or mirror-image parts (radial or bilateral) & 40
(267) & TRY database (Kattge et al. 2020)\tabularnewline
\bottomrule
\end{longtable}

*Number of plant species for which each trait was available (total
number of species used in the PACo analysis for cophylogenetic signal)

\textsuperscript{1}A trichromatic phenotype was simulated based on S, M
and L cones, with peak spectral sensitivities (\selectlanguage{greek}λ\selectlanguage{english}max) of 425 and 53,
respectively. The peak sensitivities selected roughly correspond to a
typical old word monkey phenotype, as well as many Strepsirrhines (Nevo
et al. 2018)

\textsuperscript{2}First and second axis explained 64\% of the variance
in nutritional content of fruits (Supplementary Material S7).

\textsuperscript{}\textbf{Phylogenetic reconstruction}

We used ultrametric time-calibrated phylogenies and reconstructed both
plant and lemur phylogeny at the species-level. For plants, we used two
phylogeny sets: (a) complete, including all plant species in our
dataset, and (b) native, excluding plant species considered introduced
and/or invasive, following Kull et al. (2012) (SM Table S4). After
updating the plant taxonomy nomenclature (WFO 2022), species
relationships were obtained using `V.Phylomaker' package in R, the most
extensive phylogeny reconstruction tool covering the highest below
family-level plant diversity, and with accurate and updated age
estimates for internal nodes (Jin and Qian 2019). Lemur phylogeny was
obtained from Bray et al. (2019), and taxonomic resolution was
supplemented with additional references (Pastorini et al. 2001; Markolf
et al. 2013; Lei et al. 2017).

\textbf{Cophylogenetic signal (CS)}

To test for a CS between Angiosperms and lemurs, we implemented a
Procrustean Approach to Cophylogeny (PACo; Balbuena et al. 2013). The
PACo approach assesses phylogenetic congruence by optimizing the
topological fit of two phylogenies using interaction graphs derived from
the matrix of interactions (see Suplementary Material S6 for further
details). It quantifies topological similarity by the extent to which
each internal node and branch-length in a phylogenetic tree maps to a
corresponding position in the other phylogenetic tree (Blasco-Costa et
al. 2021). When co-phylogenetic signal is high, diversification patterns
of associated organisms over evolutionary time tend to be dependent, and
congruence (i.e. topological similarity) between the phylogenies of
interacting clades is expected to occur.

We calculated one CS value for each interaction type in our dataset:
frugivory, folivory, florivory, nectarivory and granivory. PACo returns
a quantification of the global fit based on observed interactions as the
sum of squared residual distances (\emph{SS} = [?]\emph{r}
\textsuperscript{2}) between phylogenetic-interaction graphs (Balbuena
et al. 2013). As in any regression analysis, the smaller the residual
distances (Procrustean residuals), the better the fit of the two
phylogenies, and the more support for a hypothesis of CS (Hutchinson et
al. 2017). CS values were considered to be significant whenever it was
smaller than 95\% of the values obtained from 1000 randomizations of
each interaction matrix. The randomization approach implemented in PACo
(and other global-fit methods) maintains the topology of the phylogeny
of each group while shuffling the associations (i.e., interactions)
between species to generate random instances of the observed data
(Balbuena et al. 2013, Hutchinson et al. 2017). To ease interpretation
of results, CS was assessed as the Procrustes coefficient of
determination (\emph{R\textsuperscript{2}} = 1 -- \emph{SS} ) (Legendre
\& Legendre 2012), following Soares et al. (2023). Thus, congruence
expresses the extent to which each node in the phylogenetic tree of
lemurs corresponds to a position in the phylogenetic tree of plants. A
perfect congruence is obtained when \emph{R\textsuperscript{2}} = 1
(i.e., \emph{SS} = 0), and can be interpreted as a strong cospeciation
evidence, which may or may not result from coevolutionary mechanisms.

\textbf{Drivers of CS}

We estimated the phylogenetic signal in species interactions (PSI)
associated to the calculation of CS to investigate the main drivers of
the observed cophylogeny and ask whether closely related species
interact with similar sets of partners. Following Russo et al. (2018) we
used Mantel tests to assess if closely related lemur or plant species
were more likely to interact with similar set of partners than expected
by chance (e.g. Kahnt et al. 2019). For each of the interacting clades,
the distance matrices compared were the between-species phylogenetic
distance and the between-species dissimilarity (Jaccard distances) in
interactions with the other clade. A significant CS (i.e. overall
phylogenetic congruence) as well as a significant PSI for both lemur and
plants were interpreted as a case of coevolution. Instead, if
phylogenies were congruent but PSI was only significant for one of the
interacting clades, this would indicate phylogenetic tracking. Finally,
a significant CS but non-significant PSI for both lemurs and plants
would suggests that the interactions were little important for the
evolution of both clades, highlighting the importance of vicariance or
other evolutionary processes. We used the function
`phylosignal\_network' from the \emph{R} -package RPANDA to estimate PSI
values and estimate significance through permutations (Perez-Lamarque et
al. 2022). In order to preserve any influence of the degree distribution
of species, we used a conservative approach and set the permutation
argument to `nbpartners'. This keeps constant the number of partners per
species and shuffle at random their identity.

\textbf{Quantitative cophylogenetic spatial framework}

To obtain a deeper understanding of the coevolution of interacting
clades, we asked whether speciation in one of the partners influenced
morphology in the other. To this purpose, we estimated distance matrices
based on morphological traits for both plant and lemur species. Next, we
superimpose each matrix with the phylogenetic distances of the
interacting partner (plants when morphology was from lemurs, and lemurs
when morphology was from plants) using a procrustean approach
(Blasco-Costa et al. 2021). Coordinates derived from principal component
analyses (PCA) were used to compute morphological distances between
species for lemurs and plants (Supplementary Material S7). For plants,
we only had complete data for fruit and seed length and width, so we
restricted the analyses to frugivory interactions, while for lemurs we
had body mass, body length, and tail length for all the species.
Congruence values were considered to be significant whenever it was
smaller than 95\% of the values obtained from 1000 randomizations of
each interaction matrix. A significant congruence of plant phylogeny
with lemur morphology would suggest that plant speciation determined
lemur body size, while a significant congruence of lemur phylogeny with
plant morphology would be more consistent with the view that lemur
speciation determined fruit and seed sizes.

\textbf{Individual contributions to the CS}

To test whether the obtained complementary information on lemur and
plant traits influence the magnitude of the CS, we fitted linear mixed
models with a lognormal distribution, using a Bayesian approximation
(details in Supplementary Material S9). We fitted separated models for
lemurs and plants, and separated models for each interaction type where
CS was significant (see Supplemental Material for details).

For lemur traits, we had complete information for all lemur species
included in our analyses. We fitted four models, one for each
interaction type: frugivory, folivory, florivory and granivory. In each
model, we included activity pattern (diurnal, nocturnal, cathemeral) and
feeding guild (nectarivorous, omnivorous, folivorous-frugivorous,
folivorous, frugivorous, insectivorous) as independent variables, and
Procrustean residuals calculated by the PACo function as response
variables. Abundant species tend to be generalists, so to account for
the variation in geographic distribution area among lemur species, we
used geographic range as a proxy of their density, given the lack of
abundance data for each specific site. This approach is supported by
strong associations between species distribution ranges and population
sizes (Estrada et al. 2017). Data on lemur distribution range (in
km\textsuperscript{2}) were extracted from IUCN maps (IUCN 2022) and
added as random effect to models. To account for the possible influence
of sample size of interactions, the total number of studies reporting
each interaction was incorporated as a random effect to the models. To
control for the possible influence of phylogenetic relatedness, we added
lemur phylogeny as a covariate in our models.

For plant traits, we had information for only few species, and each
trait was available for different sets of plants (Table 1). In order to
take better advantage of the available information, we fitted separated
models for each trait, correlating it to the respective interaction
type. To account for the possible influence of sample size of
interactions, the total number of studies reporting each interaction was
considered as random effect in the models. To control for the possible
influence of phylogenetic relatedness, we added plant phylogeny as a
covariate in our models.

\textbf{RESULTS}

Our analyses of mutualistic and antagonistic interactions between lemurs
and plants showed co-phylogenetic signals (CS), providing evidence for
phylogenetic congruence between clades. The co-phylogenetic signal was
generally low, but significant, for all interaction types (frugivory,
folivory, florivory and granivory) except nectarivory, regardless of
whether the interaction was antagonistic or mutualistic (Table 2). The
pattern also held true when removing introduced and/or invasive plant
species (SM Table S8), suggesting either a deep conservatism of
interactions or that invasion rates were low in Madagascar. In all
interaction types where we found significant CS, the contribution of
lemurs to CS was determined by both feeding guild and activity pattern.
In all cases, insectivores and nocturnal lemurs showed the largest
Procrustean residuals, and contributed the least to the CS (Fig. 1,
Supplementary Material S9-A). As for plant traits, we found little
evidence that fruit, leaf and flower traits defined individual
contribution of plants to the CS (Supplementary Material S9-B).\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/image1/image1}

\end{center}
\end{figure}

\textbf{Figure 1} -- Contribution of each lemur genus to the overall
cophylogenetic signal in (A) frugivory, (B) folivory, (C) florivory, and
(D) granivory, measured as Procrustean residuals. Colors highlight
differences among (1) feeding guilds and (2) activity pattern.
Silhouettes were built by the authors. Procrustean residuals measure the
variation in the topological fit that is not explained by the
cophylogenetic structure of the interaction matrix. Interactions with
small values contribute the most to CS, whereas those deviating more
from the shared phylogenetic history contribute the less to CS.

To more accurately demonstrate coevolution, we also need evidence that
phylogenetic distances and the dissimilarity between species in sets of
interacting partners co-vary for both plants and lemurs (PSI). Contrary
to our expectation of the existence of phylogenetic signal in species
interactions for both plants and lemurs, we found that cophylogenetic
patterns observed for frugivory, florivory and granivory emerged from
similar lemurs (in terms of the between-species dissimilarities in sets
of interacting partners) interacting with phylogenetic-related plants,
with no reciprocal trend (i. e. plant diversification was unaffected by
lemurs). In the case of folivory, however, phylogenetic congruence was
associated with significant phylogenetic signals for both plants and
lemurs. Nectarivores were the only group that did not show any evidence
of PSI for either plants or lemurs (Table 2).

Although the observed cophylogenetic patterns for frugivory do not
appear to reflect strong coevolution, we further investigated this
possibility by taking advantage of the existence of functionally
relevant morphological traits for both plants and lemurs. We found that,
in addition to the congruence between plant and lemur phylogenies
(significant CS), there was also congruence between lemur phylogeny and
plant morphology, and between plant phylogeny and lemur morphology
(Table 2). This result suggests that, to some extent, plant speciation
shaped lemur morphology (notably body size), at the same time that lemur
speciation somehow affected diaspore morphology (fruit and/or seed
sizes).

\textbf{Table 2} -- Cophylogenetic signal (CS) and Phylogenetic signal
in the interactions (PSI) between plants and lemurs\selectlanguage{english}
\begin{longtable}[]{@{}llllllllllllllll@{}}
\toprule
\begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Interaction type\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Cophylogenetic signal (CS)\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Cophylogenetic signal (CS)\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Cophylogenetic signal (CS)\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Cophylogenetic signal (CS)\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Cophylogenetic signal (CS)\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Cophylogenetic signal (CS)\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Cophylogenetic signal (CS)\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Cophylogenetic signal (CS)\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Cophylogenetic signal (CS)\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Cophylogenetic signal (CS)\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Cophylogenetic signal (CS)\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Phylogenetic signal in interactions (PSI)*\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Phylogenetic signal in interactions (PSI)*\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Phylogenetic signal in interactions (PSI)*\strut
\end{minipage} & \begin{minipage}[b]{0.04\columnwidth}\raggedright\strut
Phylogenetic signal in interactions (PSI)*\strut
\end{minipage}\tabularnewline
\midrule
\endhead
\begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
\# lemur spp.\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
\# plant spp.\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
SS\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
SS\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
SS\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
R\textsuperscript{2}\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
R\textsuperscript{2}\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
R\textsuperscript{2}\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
p-value\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
p-value\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
p-value\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
Plants\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
Plants\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
Lemurs\strut
\end{minipage} & \begin{minipage}[t]{0.06\columnwidth}\raggedright\strut
Lemurs\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
CS\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
L\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
P\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
CS\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
L\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
P\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
CS\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
L\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
P\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
Mantel correlation\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
p-value\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
Mantel correlation\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
p-value\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
Frugivory\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
40\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
735\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.99\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.99\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.99\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.01\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.01\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.01\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
\textbf{0}\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
\textbf{0}\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
\textbf{0}\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.03\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
\textbf{0.001}\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.20\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.73\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
Folivory\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
44\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
679\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.98\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.02\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
\textbf{0}\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.03\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
\textbf{0.001}\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.13\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
\textbf{0.001}\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
Florivory\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
38\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
269\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.96\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.04\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
\textbf{0}\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.04\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
\textbf{0.001}\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.06\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.57\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
Nectarivory\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
17\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
40\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.89\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.11\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.2\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.04\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.12\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-0.18\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.99\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
Granivory\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
13\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
53\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.86\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.14\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
\textbf{0}\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
-\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.20\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
\textbf{0.001}\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.07\strut
\end{minipage} & \begin{minipage}[t]{0.04\columnwidth}\raggedright\strut
0.09\strut
\end{minipage}\tabularnewline
\bottomrule
\end{longtable}

\emph{SS} -- sum of squares

\emph{R\textsuperscript{2}} -- Procrustes coefficient of determination

CS -- cophylogenetic signal (congruence between plant and lemur
phylogenies)

L -- lemur interactions (congruence of lemur phylogeny with differences
in plant morphology)

P -- plant interactions (congruence of plant phylogeny with differences
in lemur morphology

*Interpretation of PSI: a p-value \textgreater{} 0.05 for plants means
that similar lemurs tend not to interact with closely related plant
species. Similarly, a p-value \textgreater{} 0.05 for lemurs means that
similar plants tend not to interact with closely related lemurs. If
p-values for both plants and lemurs are \textgreater{} 0.05 (as is the
case of nectarivory), it means that there is no significant phylogenetic
signal in species interactions.

\textbf{DISCUSSION}

For a deeper understanding of the potential coevolution between lemur
and plants, we tested for cophylogenetic signals in plant-lemur
interactions, and detected significant, although weak, signals in
frugivory, folivory, florivory and granivory interactions. In the case
of frugivory, plant speciation seems to have influenced lemur body size,
at the same time that lemur speciation modulated diaspore size. Weak
cophylogenetic signals do not necessarily imply lack of evolutionary
history shared between symbiotic partners. Indeed, cophylogenetic
patterns observed for frugivory, florivory and granivory emerged from
lemurs sharing similar sets of partners interacting with
phylogenetically-related plants with no reciprocal trend. In the case of
folivory, plant diversification was also affected by lemurs. Our results
do not give support to a strong coevolutionary diversification between
lemurs and plants, in agreement with previous observations for
Neotropical primates and Angiosperms (Fuzessy et al. 2022). Instead, our
findings reflect that lemur feeding behavior is very generalist,
specially of large lemur species, and support the idea that
cophylogenetic signals are modulated by processes that are not directly
related to strong co-speciation events, such as vicariance, phylogenetic
tracking, and diffuse coevolution (Blasco-Costa el al. 2021).

The role of vicariance in coevolution ---where common biogeographic
events cause subdivision of distribution ranges of interacting partners
---is dependent on the geographic scale. Vicariance contributes to
coevolutionary processes by promoting diversification of entire
populations that become isolated from each other. Over time, as
populations diverge and adapt to their respective environments and
evolve distinct and specialized structures and traits, including those
that are important for their close relationship, they become less
compatible. In this sense, vicariance is associated with high
cophylogenetic signals (Blasco-Costa el al. 2021), so in the specific
case of Madagascar, especially in the context of a relatively small
island, it is not expected to play a significant role. Phylogenetic
tracking is also usually associated with high cophylogenetic signals. It
refers to a tendency of interacting partners to track each other's
evolutionary history, such that closely related species are more likely
to interact than distantly related species (Blasco-Costa et al. 2021).
Despite our finding of low cophylogenetic signals in lemur-plant
interactions, phylogenetic congruence arose even in a supergeneralist
system where other plant-consuming animals, such as insects and other
vertebrates, may also be shaping plant diversification. The patterns
observed here for frugivory, florivory and granivory emerged from
similar lemurs interacting with closely related plants with no
reciprocal trend (i.e., plant diversification was unaffected by lemurs).
In this case, generalist lemurs seem to be tracking temporal
fluctuations in resource availability, and increasing the consumption of
one plant species at the expense of decreased consumption of an
alternative, phylogenetically-related, resource, known as switching
behavior (Carnicer et al. 2008). Such low levels of CS associated with a
correlation between phylogenetic distances and dissimilarities in sets
of interacting plants for lemurs can be interpreted as an indicative
case of diffuse coevolution. In this sense, each lemur species interacts
with a set of plants, and experiences a complex suite of selective
pressures derived from the set of plant species they interact with.

In the case of folivory, similar plants were also found to interact with
closely related lemur species, as an indicative of coevolution. Except
for the strict folivorous genera \emph{Hapalemur} and \emph{Lepilemur} ,
lemurs use a variety of resources as feeding items, such as fruits,
leaves flowers, seeds, nuts, nectar, and animal prey. Regardless of the
main resource, leaves constitute an important resource for most species,
reinforcing the idea that lemur-plant interactions are quite generalist.
A weak cophylogenetic signals associated with positive correlations
between phylogenetic distances and dissimilarities in sets of
interacting partners for both lemurs and plants may also be interpreted
as a case of diffuse coevolution, since distinct amounts of each feeding
item vary across taxa, and other coexisting frugivorous and herbivorous
clades act together, thus, favoring a diffuse coevolutionary process
(Erikson 2016).

The multi-specific nature of the process delineating cophylogenetic
patterns, as observed here, does not necessarily mean either that lemurs
and plants have not coevolved, or that coevolution has necessarily been
the primary force fueling diversification (e.g. Althoff et al. 2014;
Fuzessy et al. 2022). Taken together, our results agree with the notion
that vertebrates and plants can influence each other evolution but also
highlight that they do not share a very tight coevolutionary history
(Valenta \& Nevo 2020, Fuzessy at al. 2022), such as those observed in
host-parasite interactions (Gandon \& Michalakis 2002) or in some
plant-pollinator systems (Herrera 2019). Spatio-temporal asymmetries,
disruptions in the strength of the relationships between interacting
partners, and shifts between periods of coevolution and independent
evolution seem to be leading to adaptive changes, ultimately resulting
in a weak and continuous process (Erikson 2016), in agreement with
previous observations in the Neotropics (Fuzessy et al. 2022). The
resemblance between Madagascar and the Neotropics is remarkable, given
that the number of frugivorous mammals in Madagascar is low when
compared to other tropical communities, which are dominated for
frugivorous primates (Fleming et al. 1987; Goodman \& Ganzhorn 1997). In
Madagascar, there are very few medium-sized strictly frugivorous lemurs
(mostly \emph{Varecia} and some species of \emph{Eulemur} ), and the
proportion of fruits consumed by Malagasy lemurs is low compared with
primates in other regions (Fleagle \& Reed 1996). The distinctive
phenology of Malagasy forests, associated with a great variability in
spatio-temporal fruit availability has been proposed as the main drivers
of the underrepresentation of frugivores in Madagascar (Dewar \& Richar
2007). These patterns highlight potential risks associated with the
current disruption of lemur-plant interactions. Despite high levels of
generalism, we have shown that extant lemur-plant interactions are
phylogenetically constrained, so if a species belonging to an already
limited community of frugivores is lost, the probability of other
species to fulfill the vacant role is reduced. This issue may be
exacerbated if the species lost play the role of both dispersers and
pollinators, as is the case between the black-and-white ruffed
lemur,\emph{Varecia variegata} , and the traveler palm, \emph{Ravenala
madagascariensis} (Kress et al. 1994). Such double mutualisms appear to
be highly prevalent in island ecosystems, and pose additional risks for
the continuing health of these forests when facing the extinction of
frugivorous animal partners (Fuster et al. 2019).

Overall, lemurs consumed plant species that were more closely related
than expected by chance, potentially indicating the consumption of
plants with similar traits. In addition, invasive plant species have not
disrupted the cophylogenetic structure of the interaction networks,
suggesting a deep conservatism of interactions (or a low invasion rate
in Madagascar). Indeed, we found no clear pattern of the influence of
plant traits on the cophylogenetic signals. Nevertheless, to make any
precise statement in this regard, we need complete and more detailed
information on plant traits, including those related to fruits, leaves,
and flowers, which are most frequently consumed by lemurs. Therefore, we
encourage future studies to characterize Malagasy flora in terms of
functional characteristics related to interactions with mutualistic and
antagonistic partners (e. g. Albert-Daviaud et al. 2021).

Lemur feeding preferences and activity patterns likely shaped the
congruence between their phylogeny with that of plants. Due to the
unique ecology of Madagascar, lemurs evolved to occupy a wide range of
niches, from night-living arboreal insectivores to ground-dwelling
cathemeral folivores-frugivores, and diurnal frugivores (Gould \&
Sauther 2006). As expected, nocturnal lemurs and those that include a
great amount of insects in their diets (and the least amount of plants),
showed the largest Procrustean residuals, and contributed the least to
the shared evolutionary history with plants, regardless of interaction
type, and in agreement with previous studies (Fuzessy et al. 2022).
Overall, diet and activity patterns have been important drivers of lemur
evolution, shaping morphology, physiology, and behavior in unique ways
that have allowed them to thrive in Malagasy diverse habitats. Dietary
adaptations have driven the evolution of morphological and physiological
features, such as specialized teeth, tongue, digestive systems, and
metabolic rates (Cuozzo \& Yamashita 2006; Powzyk \& Mowry 2006; Pastor
et al. 2021). Over evolutionary time, insectivores such
as\emph{Daubetonia} , for instance, have evolved specialized teeth and
long fingers that are adapted for catching and handling prey, added to
an enhanced night vision and specialized auditory systems to aid in
hunting (Goodman et al. 1993; Sterling \& McCreless 2006; Lhota et al.
2008). On the other hand, lemurs that consume great amounts of fruits
and flowers have evolved color vision to help them distinguish ripe
fruits and identify flowers, specially from a conspicuous background
(Nevo et al. 2018).

Our study highlighted the important role of evolutionary history in
structuring modern interaction patterns between vertebrates and plants.
Our cophylogenetic analysis compared phylogenies of interacting partners
based on observed extant interactions presented as binary lemur-plant
interaction matrices. While this approach provides an important basis
for investigating cophylogenetic relationships (Balbuena et al. 2013;
Blasco-Costa et al. 2021), it ignores that the strength of associations
among interacting partners may affect evolutionary outcomes, and thus
precludes a deeper assessment of cophylogenetic processes. In the case
of frugivory interactions, in addition to the congruence between plant
and lemur phylogenies, we also detected a congruence between lemur
phylogeny and plant morphology, and between plant phylogeny and lemur
morphology. Although this result suggests that, to some extent, plant
speciation shaped lemur body size, and lemur speciation somehow affected
diaspore morphology, we still lack sufficient information on the role of
plant traits, including those related to fruits, leaves, and flowers, in
shaping the cophylogenetic patterns described. Studies characterizing
Malagasy flora in terms of functional characteristics related to
interactions with mutualistic and antagonistic partners are urgently
required (e. g. Albert-Daviaud et al. 2021). Despite these limitations,
results presented here shed unprecedented light on the potential for
supergeneralist interactions, including both antagonisms and mutualisms,
to shape shared evolutionary trajectories between vertebrates and
Angiosperms.

\textbf{REFERENCES}

Albert-Daviaud, A., Perillo, S. \& Stuppy, W. (2021). Data from: Seed
dispersal syndromes in the Madagascan flora: The unusual importance of
primates. Dryad Digital Repository. Available
at:\emph{https://doi.org/10.5061/dryad.dt940}

Althoff, D.M., Segraves, K.A. \& Johnson, M.T.J. (2014) Testing for
coevolutionary diversification: linking pattern with
process.\emph{Trends Ecol Evol} , 29, 82--89

Balbuena, J. A., M\selectlanguage{ngerman}íguez-Lozano, R. \& Blasco-Costa, I. (2013). PACo: A
Novel Procrustes Application to Cophylogenetic Analysis. \emph{PLoS ONE}
, 8(4), e61048.\emph{https://doi.org/10.1371/journal.pone.0061048}

Blasco-Costa, I., Hayward, A., Poulin, A. \& Balbuena, J.A. (2021).
Next-generation cophylogeny: unravelling ecoevolutionary processes
.\emph{Trends Ecol Evol} , 36(10),
907-918.\emph{https://doi.org/10.1016/j.tree.2021.06.006}

Bray, J., Amoroso, C.R., Vining, A.Q. \& Nunn, C.L.
(2023).\emph{10kTraits: A Database for Comparative Primatology} . Beta
release.

\emph{http://10kTrees.fas.harvard.edu}

Brodie, E. D. \& Ridenhour, B. J. (2003). Reciprocal selection at the
phenotypic interface of coevolution. \emph{Integr Comp Biol} , 43,
408-418.

Callmander, M.W., Phillipson, P.B., Schatz, G.E., Andriambololonera, S.,
Rabarimanarivo, M., Rakotonirina, N. et al. (2011). The endemic and
non-endemic vascular flora of Madagascar updated. \emph{Plant Ecol Evol}
, 144, 121--125

Carnicer, J., Abrams, P. A. \& Jordano, P. (2008). Switching behavior,
coexistence and diversification: comparing empirical community-wide
evidence with theoretical predictions. \emph{Ecol Lett} , 11(8),
802--808.

Cuozzo, F. P. \& N. Yamashita. (2006). Impact of ecology on the teeth of
extant lemurs: a review of dental adaptations, function, and life
history. In: \emph{Lemurs: ecology and adaptation} , eds. Gould, L. \&
Sauther, M. L. Springer, New York.

Dewar, R. E. \& Richard, A. F. (2007). Evolution in the hypervariable
environment of Madagascar. \emph{PNAS} , 104 (34), 13723-13727.

Dewar, R. E. \& Richard, A. F. (2012). Madagascar: A history of
arrivals, what happened, and will happen next. \emph{Annu Rev Anthropol}
, 41, 495--517

Díaz, S., Kattge, J., Cornelissen, J.H.C., Wright, I. J., Lavorel, S.,
Dray, S. et al. (2022). The global spectrum of plant form and function:
enhanced species-level trait dataset. \emph{Sci Data} , 9, 755.

\emph{https://doi.org/10.1038/s41597-022-01774-9}

Dismukes, W., Braga, M. P., Hembry, D. H., Heath, T. A. \& Landis, M. J.
(2022). Cophylogenetic Methods to Untangle the Evolutionary History of
Ecological Interactions. \emph{Ann Rev Ecol Syst} , 53, 275-298

Eriksson, O. (2016) Evolution of angiosperm seed disperser mutual- isms:
the timing of origins and their consequences for coevo- lutionary
interactions between angiosperms and frugivores. \emph{Biological
Reviews} , 91, 168--186.

Fleagle, J. G. \& Reed, K. E. (1996). Comparing primate communities: a
multivariate approach. \emph{J Human Evol} , 30, 489-510.

Fleming, T.R., Breitwisch, R. \& Whitesides, G. H. (1987). \emph{Annu
Rev Ecol Syst} , 18, 91--109.

Fuster, F., Kaiser\selectlanguage{english}-Bunbury, C., Olesen, J.M. \& Traveset, A. (2019).
Global patterns of the double mutualism phenomenon. \emph{Ecography} ,
42, 826--835.

Fuzessy, L.F, Silveira, F.A.O., Culot, L., Jordano, P \& Verdu, M.
(2022). Phylogenetic congruence between Neotropical primates and plants
is driven by frugivory. \emph{Ecol Lett} , 25, 320-329.

Galan-Acedo, C., Arroyo-Rodriguez, V., Andresen, E. \& Arasa-Gisbert, R.
(2019). Ecological traits of the world's primates. \emph{Sci Data} , 6
(55). \emph{https://doi.org/10.1038/s41597-019-0059-9}

Gandon, S. \& Michalakis, Y. (2002). Local adaptation, evolutionary
potential and host--parasite coevolution: interactions between
migration, mutation, population size and generation time. \emph{J Evol
Biol} , 15, 451-462.

Genin, F., Mazza, P. P., Pellen, R., Rabineau, M., Aslanian, D. \&
Masters, J.C. (2022). Co-evolution assists geographic dispersal: the
case of Madagascar. \emph{Biol J Linn Soc} , 137, 163--182.

Goodman, S. M. \& Ganzhorn, J. U. (1997). Rarity of figs
\emph{(Ficus)}on Madagascar and its relationship to a depauperate
frugivore community.\emph{Revue d'Ecologie, Terre et Vie,} 52 (4),
321-329.

Goodman, S. M., Andrianarimisa, A., Armstrong, A. H., Cooke, A., De Wit,
M., Ganzhorn, J. U., Gautier, L., Goodman, S. M., et al.
(2022).~\emph{The New Natural History of Madagascar} . Princeton
University Press. https://doi.org/10.2307/j.ctv2ks6tbb

Goodman, S.M. \& Benstead, J.P. (2005). Updated estimates of biotic
diversity and endemism for Madagascar. \emph{Oryx} , 39, 73--77.

Goodman, S.M., O~\selectlanguage{english}\selectlanguage{english}Connor, S. \& Langrand, O. (1993). A review on
predation on lemurs: implications for the evolution of social behavior
in small, nocturnal primates. In: \emph{Lemur social system and their
ecological basis} , eds P.M. Kappeler, P. M. \& Ganzhorn J. U. Plenum
Press, New York, pp. 51-66

Gould, L. \& Sauther M. L. (2006). \emph{Lemurs. Developments in
Primatology: Progress and Prospect} . Springer, Boston, MA.

Hembry, D. H. \& Weber, M. G. (2020). Ecological interactions and
macroevolution: a new field with old roots. \emph{Annu Rev Ecol Syst} ,
51, 215-243

Hembry, D. H., Yoder, J. B. \& Goodman, K. R. (2014). Coevolution and
the Diversification of Life. \emph{The Amer Naturalist} , 184, 425--438.

\emph{https://doi.org/10.1086/677928}

Herrera, C. M. (2019). Flower traits, habitat, and phylogeny as
predictors of pollinator service: a plant community
perspective.\emph{Ecol Monograph} , 90, e01402.

Hutchinson, M.C., Cagua, E.F. \& Stouffer, D.B. (2017) Cophylogenetic
signal is detectable in pollination interactions across ecological
scales. \emph{Ecology} , 98(10), 2640--2652. https://doi.
org/10.1002/ecy.1955

IUCN. 2022. \emph{The IUCN Red List of Threatened Species} . Version
2022-1. https://www.iucnredlist.org. Accessed on 1 September 2022.

Johnson, C.A., Smith, G.P., Yule, K. et al. (2021). Coevolutionary
transitions from antagonism to mutualism explained by the Co-Opted
Antagonist Hypothesis. \emph{Nat Commun} , 12, 2867.

Kahnt, B., Hattingh, W. N., Theodorou, P., Wieseke, N., Kuhlmann, M.,
Glennon, K. L., et al. (2019) Should I stay or should I go? Pollinator
shifts rather than cospeciation dominate the evolutionary history of
South African \emph{Rediviva} bees and their \emph{Diascia} host
plants.\emph{Mol Ecol} , 28, 4118--4133

Kattge, J, Boenisch, G, D\selectlanguage{ngerman}íaz, S, et al. (2020). TRY plant trait database
- enhanced coverage and open access. \emph{Glob Change Biol} , 26:
119-188. \emph{https://doi.org/10.1111/gcb.14904}

Kress, W.J., Schatz, G.E., Andrianifahanana, M. \& Morland, H.S. (1994).
Pollination of \emph{Ravenala madagascariensis} (Strelitziaceae) by
lemurs in Madagascar: Evidence for an archaic coevolutionary
system?\emph{Am J Bot} , 1994, 81, 542--551

Kull, C.A., Tassin, J., Moreau, Ramiarantsoa, H. R., Blanc-Pamard, C. \&
Carrière, S. M. (2012).~The introduced flora of
Madagascar.~\emph{Biological Invasions} ~14, 875--888.

\emph{https://doi.org/10.1007/s10530-011-0124-6}

Laine, A-L. 2009. Role of coevolution in generating biological
diversity: spatially divergent selection trajectories. \emph{Journal of
Experimental Botany} , 60, 2957--2970.

Legendre, P. \& Legendre, L. (2012). \emph{Numerical Ecology} . Third
English Edition. Amsterdam: Elsevier.

Lei, R., Frasier, C. L., Hawkins, M. T. R., Engberg, S. E. Bailey, C.
A., et al. (2017). Phylogenomic Reconstruction of Sportive Lemurs (genus
Lepilemur) Recovered from Mitogenomes with inferences for Madagascar
biogeography. \emph{Journal of Heredity} , 108, 107--119.
https://doi.org/10.1093/jhered/esw072

Lhota, S., J\selectlanguage{polish}ů\selectlanguage{english}nek, T., Bartos, L. \& Kub\selectlanguage{polish}ĕ\selectlanguage{english}na, A.A. (2008). Specialized use
of two fingers in free-ranging aye-ayes (\emph{Daubentonia
madagascariensis} ). \emph{Am J Primatol} , 70(8), 786-95.

Markolf, M. \& Kappeler, P.M. (2013). Phylogeographic analysis of the
true lemurs (genus Eulemur) underlines the role of river catchments for
the evolution of micro-endemism in Madagascar. \emph{Front Zool} , 10,
70. \emph{https://doi.org/10.1186/1742-9994-10-70}

Medina, M., Baker, D.M., Baltrus, D.A,. Bennett G.M., Cardini, U.,
Correa, A.M.S., et al. (2022). Grand Challenges in
Coevolution.\emph{Front Ecol Evol} , 9, 618251.

Mittermeier, R. A., Louis Jr., E. E., Richardson, M., Schwitzer, C.,
Langrand, O. \& Rylands, A. B. (2010). \emph{Lemurs of Madagascar}(3ed).
Conservation International, Arlington, USA.

Nevo, O., Valenta, K., Razafimandimby, D., Melin, A.D., Ayasse, M. \&
Chapman, C. A. (2018). Frugivores and the evolution of fruit
colour.\emph{Biol Lett} , 142018037720180377

Nilsson, L. A. (1988). The evolution of flowers with deep corolla
tubes.\emph{Nature} , 334, 147-149.

Norconk, M. A. (2020). Historical antecedents and recent innovations in
pitheciid (titi, saki, and uakari) feeding ecology. \emph{Am J Primatol}
, e23177.

Nuismer, S. L., Jordano, J. \& Bascompte, J. (2013). Coevolution and the
architecture of mutualistic networks. \emph{Evolution} , 67, 338--354

Pastor, J.F., Muchlinski, M.N., Potau, J.M., Casado, A., Garc\selectlanguage{ngerman}ía-Mesa,
Y., Vega, J.A. \& Cabo, R. (2021). The Tongue in Three Species of
Lemurs: Flower and Nectar Feeding Adaptations. \emph{Animals} , 11,
2811.

\emph{https://doi.org/10.3390/ani11102811}

Pastorini, J., Forstner, M.R.J. \& Martin, R.D. (2001). Phylogenetic
history of sifakas (Propithecus: Lemuriformes) derived from mtDNA
sequences. \emph{Am J Primatol} , 53, 1--17.

Perez-Lamarque, B., Maliet, O., Pichon, B., Selosse, M.-A., Martos, F.
\& Morlon, H. (2022). Do closely related species interact with similar
partners? Testing for phylogenetic signal in bipartite interaction
networks. \emph{Peer Community Journal} , 2, e59.

Powzyk, J.A. \& Mowry, C.B. (2006). The Feeding Ecology and Related
Adaptations of \emph{Indri indri} . In: \emph{Lemurs. Developments in
Primatology: Progress and Prospect} s, eds. Gould, L., \& Sauther, M.L.
Springer, Boston, MA.

Pozzi, L., Hodgson, J. A., Burrell, A. S., Sterner, K. N., Raaum, R. L.
\& Disotell, T. R. (2014). Primate phylogenetic relationships and
divergence dates inferred from complete mitochondrial
genomes.\emph{Molecular Phylogenetics and Evolution} , 75, 165--183.

\emph{https://doi.org/10.1016/j.ympev.2014.02.023}

Razafindratsima, O. H, Yacoby, Y. \& Park, D.S. (2018). MADA: Malagasy
Animal trait Data Archive. \emph{Ecology} , 99 (4), 990-990.

Richard, A.F. \& Dewar, R.E. (1991). Lemur ecology. \emph{Annu Rev Ecol
Syst} 22, 145--175.

Russo, L., Miller, A.D., Tooker, J., Bjornstad, O.N. \& Shea, K. (2018).
Quantitative evolutionary patterns in bipartite networks: vicariance,
phylogenetic tracking or diffuse coevolution? \emph{Methods Ecol Evol} ,
9, 761--772

Schupp, E. W., Jordano, P. \& Gómez, J. M. (2010). Seed dispersal
effectiveness revisited: a conceptual review. \emph{New Phytol} , 188,
333-353.

Soares, G., Adriano, E., Domingues, M. \& Balbuena, J.A. (2023).
Diversification processes between monogenoids (Dactylogyridae) and their
marine catfish (Siluriformes: Ariidae) from the Atlantic coast of South
America. \emph{Parasitology} , 150, 184-194.
doi:10.1017/S0031182022001615

Sterling, E.J. \& McCreless, E.E. (2006). Adaptations in the Aye-aye: A
Review. In: \emph{Lemurs. Developments in Primatology: Progress and
Prospect} s, eds. Gould, L., \& Sauther, M.L. Springer, Boston, MA.

Strauss, S. Y. \& Irwin, R. E. (2004). Ecological and Evolutionary
Consequences of Multispecies Plant-Animal Interactions. \emph{Annu Rev
Ecol Syst} , 35, 435--466. \emph{http://www.jstor.org/stable/30034123}

WFO (2022): \emph{World Flora Online} . Version 2022.11. Published on
the Internet; http://www.worldfloraonline.org. Accessed on: 11/11/2022

Thompson, J. N. (1994). The coevolutionary process. The University of
Chicago Press. Chicago, USA.

Valenta, K. \& Nevo, O. (2020) The dispersal syndrome hypothesis: how
animals shaped fruit traits, and how they did not. \emph{Functional
Ecology} , 34, 1158--1169.

Valenta, K., Nevo, O. \& Chapman, C.A. (2018) Primate fruit color:
useful concept or alluring myth? \emph{International Journal of
Primatology} , 39(3), 321--337. https://doi.
org/10.1007/s10764-018-0025-y

Yoder, J.B. \& Nuismer, S.L. (2010). When does coevolution promote
diversification? \emph{Am Naturalist} , 176, 802--817\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Figure1/Figure1}

\end{center}
\end{figure}

\selectlanguage{english}
\FloatBarrier
\end{document}

