Comparative morphology of populations of Monstera Adans.

(Araceae) from natural forest fragments in Northeast Brazil

using elliptic Fourier Analysis of leaf outlines
I. M. Andrade
1
, S. J. Mayo
2
, D. Kirkup
2
& C. Van Den Berg
1
Summary. A comparative study of the leaf outline morphometrics of Monstera adansonii var. klotzschiana, M. adan-
sonii var. laniata and M. praetermissa was carried out. The study focused on populations in isolated montane humid
(brejo) forests of Cear state in Northeast Brazil and compared them with populations from Amazonia and the
Brazilian Atlantic Forest. Digitised outlines were prepared from a total of 1,695 eld-collected leaf images from 20
populations, and elliptic Fourier analysis was used to generate matrices of coefcients, from which six shape
variables (principal components) were extracted using Principal Components Analysis. Intra-population variability
and inter-population differences were analysed with multivariate distance methods. Separate analyses were carried
out for each of three leaf size classes (juvenile, submature, mature) because of the strong heteroblasty typical of
this genus. Juvenile leaves were the least variable size class within populations of M. adansonii var. klotzschiana. The
shape variables expressed very similar types of variation in all three size classes. The Cear brejo populations of M.
adansonii var. klotzschiana showed signicant differences between mature leaf outlines in all pairwise comparisons;
the Pacatuba population was the most distinct. The Cear populations did not cluster together exclusively. In all
three size classes, populations clustered together into their taxonomic groups, most clearly so in mature leaves. No
correlation between morphological and geographic distance matrices was found, nor between morphological and
molecular distance. The study showed that leaf outline shape is a practicable and useful quantitative trait for
studying morphological variability at species, varietal and population levels.
Key Words. Araceae, Brazil, brejo forest, Cear, elliptic Fourier Analysis, Monstera, morphometrics.
Introduction
The context of this study is the brejo forest a
mosaic of isolated humid forest areas in Northeast
Brazil scattered within the regions predominant semi-
arid caatinga biome wherever rainfall brought by
humid winds from the Atlantic Ocean provides
suitable microclimates in mountainous terrain (Sales
et al. 1998; Rodal et al. 1998; Sampaio et al. 2002;
Borges-Nojosa & Caramaschi 2003; Prto et al. 2004;
Andrade et al. 2007). The brejo forests are today
classied as part of the Brazilian Atlantic forest in the
category of seasonal semi-deciduous montane forest
(IBGE 1985, 2005). In Cear State there are seven
such areas (Figueiredo 1997) situated roughly halfway
between Amazonia and the Atlantic forest, Brazils
major humid forest regions.
The brejo forests are of particular biogeographic
interest because they are thought to be natural fragments
representing relicts of previous expansions of the Atlantic
and Amazon forests (AbSber 1982; Andrade-Lima
1982; Borges-Nojosa & Caramaschi 2003). The results
of Oliveira et al. (1999), in a study of a Quaternary
pollen prole from central Bahia (relatively near the
focus of our study in Cear), suggest that humid and
cooler climates existed in Northeast Brazil until around
4,500 yrs BP, and they discussed two hypotheses which
postulate corridors of humid forest linking the Amazon
and Atlantic forests; one along the northeastern coast of
Brazil from Maranho to the region around Salvador in
Bahia and the other across central Brazil through a
network of gallery forests. The existence of such
connections and associated patterns of gene ow over
Accepted for publication June 2008.
1
Programa de Ps-Graduao em Botnica, Departamento de Cincias Biolgicas, Universidade Estadual de Feira de Santana (UEFS), Avenida Universitria s/n,
44031-460, Feira de Santana, Bahia, Brazil.
2
Herbarium, Royal Botanic Gardens, Kew, Richmond, Surrey TW9 3AB, UK. e-mail: s.mayo@kew.org
Electronic supplementary material The online version of this article (doi:10.1007/s12225-008-9032-z) contains supplementary material, which is available to
authorized users.
KEW BULLETIN VOL. 63: 193211 (2008)
The Board of Trustees of the Royal Botanic Gardens, Kew, 2008
wide areas of eastern South America is consistent with
recent results from lizard and amphisbaenid biogeogra-
phy (Borges-Nojosa & Caramaschi 2003), and our own
results using AFLP molecular markers in Monstera
adansonii Schott var. klotzschiana (Schott) Madison
(Andrade et al. 2007).
This paper is part of a population level study aimed at
comparing morphological and molecular patterns inindi-
vidual species of the family Araceae occurring in brejo
forests in the Brazilian state of Cear and in the Atlantic
and Amazon forests; a molecular study of Monstera
adansonii var. klotzschiana has already been published
(Andrade et al. 2007).
Monstera adansonii var. klotzschiana was chosen as a
typical representative of the herbaceous brejo ora for
three main reasons. Firstly as a root-climber of humid
tropical forests it is intolerant of low humidity and is
conned ecologically to this habitat it has not been
observed in caatinga or seasonally dry forests. On the
other hand, it is rarely found in undisturbed primary
forest, occurring typically in secondary moist tropical
forest with a well-developed canopy. Flowering is most
conspicuous where light levels are higher, at the edge
of clearings and along forest edges. We observed that
in Cear this taxon only occurs near waterfalls and
streams that are permanent throughout the year.
Second, this taxon is widespread in the eastern half
of tropical South America (Madison 1977), allowing
comparison of the Cear populations with those in
Amazonia and the Atlantic forest. It is one of several
species of Araceae which show a disjunct biogeo-
graphical range between these two major humid forest
biomes (Mayo 1983). Finally, previous work (Andrade
& Mayo 1998) provided a good understanding of the
morphology of the plant in its natural habitat.
Our aim was to estimate quantitatively morpho-
logical divergence between the populations with a view
to comparison with genetic patterns estimated with
molecular AFLP markers. Using leaf outline morpho-
metrics as data for estimating morphological variation
(see Methods for further explanation) we focusedonthe
following specic objectives: 1. Comparison of the Cear
brejo populations of Monstera adansonii var. klotzschiana;
2. Estimation of the morphological differences between
these populations; 3. Comparison of the Cear popula-
tions with those in the Atlantic and Amazon forests; 4.
Comparison of leaf outline morphology in var. klotzschi-
ana with that of the related taxa in Northeast Brazil,
M. adansonii var. laniata (Schott) Madison and M.
praetermissa E. G. Gon. & Temponi.
Materials and Methods
Leaf outline morphometrics. Two constraints led to
the choice of leaf outline data for the following
reasons. Firstly, to obtain results comparable with our
molecular study (Andrade et al. 2007), adequately
sampled morphological data from the same popula-
tions were needed in a form which could be analysed
using similar quantitative techniques. Second, limited
resources for eldwork and the large geographical
distances between study populations meant that many
localities could be sampled only once. These factors
favoured leaves since, unlike inorescences, they were
nearly always available in sufcient numbers and easily
sampled and preserved. Using a digital camera, leaf
outline shapes and dimensions were easily captured in
the eld. Digitisation of the leaf outlines from images
and obtaining mathematical descriptions (using ellip-
tic Fourier Analysis) from the resulting coordinate
data was made easy by freely available software
(TpsDig: Rohlf (2004); Morpheus et al.: Slice (2008);
SHAPE: Iwata & Ukai (2002); download from Rohlf
2006).
Taxonomic determination of Monstera species nor-
mally demands observation of characters from inor-
escences as well as vegetative organs (Madison 1977).
Fertile voucher specimens were gathered from sam-
pled populations whenever found, and identied
using the standard revision (Madison 1977); these
were deposited permanently as herbarium specimens
(see Appendix and Electronic Supplementary Material).
Combinations of vegetative characters were used to
identify eld-collected specimens of the three taxa of
Monstera studied. M. praetermissa is easily distinguished
from M. adansonii in non-owering plants as follows:
the mature plant is altogether smaller in stature; the
leaves become dark brown to blackish after drying; the
internodes are much more slender, darker green and
lack the abundant whitish speckling of M. adansonii
instead the epidermis is shallowly verrucate, the
verrucae being the same colour as the rest of the
internode. The petiole sheath margins are very quickly
marcescent, decomposing into fragile black membra-
naceous fragments which are eventually deciduous; in
M. adansonii the sheath margins are persistent, green
and herbaceous. The leaf perforations in M. praetermissa
are relatively few and large, extending almost to the
very margin of the blade, which often becomes partly
pinnatid by the rupture of the remaining marginal
strip. In M. adansonii leaf perforation patterns are very
variable, ranging from absent to extremely abundant
and with the individual perforations varying widely in
size and shape. However, they never extend right to the
very margin and the blade thus does not become
pinnatid.
Within Monstera adansonii, the differentiation of var.
klotzschiana and var. laniata is problematic in fragmen-
tary herbarium material, but easier in living eld
populations. The most reliable diagnostic characters
are in the owers: in var. klotzschiana the stigma is
194 KEW BULLETIN VOL. 63(2)
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elliptic and raised up on a shallow but distinct stylar
prominence at the gynoecium apex, whereas in var.
laniata the stigma is linear and sessile on a truncate,
attened gynoecial apex. Living non-owering plants
can be distinguished in the eld as follows: var.
klotzschiana prefers more shaded and protected hab-
itats under forest canopies and has leathery mature
leaf blades which are more than twice as long as wide
and ovate-elliptic to oblong-elliptic in shape; the leaf
blade base is less asymmetric, usually acute on one
side and obtuse to rounded on the other. Var. laniata
typically grows in more open exposed habitats such as
coastal scrub on rocks or even on fence posts in humid
tropical pasture; the mature leaf blades are rather thin
in texture, less than twice as long as wide and broadly
ovate to ovate-elliptic, with strongly asymmetric leaf
blade bases acute on one side and usually truncate to
subcordate on the other. Var. laniata is quite rare in
non-Amazonian Brazil, its main range being Central
America, Pacic tropical South America, western
Amazonia, Venezuela, the Guianas and northern
Amazonian Brazil. Madison (1977) cited no records
for eastern or northeastern Brazil, but subsequent
eldwork uncovered a form of var. laniata in the
Atlantic forest of southern Bahia and Pernambuco;
the Bahian plants are notable for their very numerous
and irregular leaf perforations.
The degree to which leaf outline shape is geneti-
cally determined is an important issue from a taxo-
nomic standpoint. The experimental studies needed
to investigate this, involving comparative growth trials
in an experimental garden, were beyond the scope of
this project. However, there is evidence which suggests
that leaf shape has a genetic basis both in general and
in the particular case of Monstera. Kessler & Sinha
(2004) reviewed studies of genetic control of leaf
shape in model organisms and discussed a range of
genetically mediated processes involved in establishing
the mature shape of the leaf blade. Studies by Furuta et
al. (1995), Yamanaka et al. (2001) and Iwata et al.
(2002) used principal component shape variables
derived from matrices of Elliptic Fourier coefcients
to describe leaf shape quantitatively in selected
cultivars and varieties of soybean and citrus. All these
studies found signicant genetic components in the
shape variables, and were furthermore able to show
that the variables which describe symmetric changes in
leaf shape have a much higher genetic component
that the asymmetrical ones. Yoshioka et al. (2004)
obtained similar results using petal shape in primula
cultivars. The heteroblasty shown by all Monstera
species also indicates a major role for genetic deter-
mination in leaf shape in these plants. Juvenile leaves
always differ considerably in shape, size and presence
or pattern of perforations and undergo a predictable
series of shape changes as the plant develops from
seedling to adult; this was used by Madison (1977) as
the basis of his sectional classication of the genus.
Leaf outline capture. 1,695 leaves were sampled from
natural populations at 32 sites in Brazil and French
Guiana, grouped into 20 populations (Fig. 1, Table 1,
Electronic Supplementary Material). At each site,
leaves at various stages of the growth cycle were
collected according to availability. To avoid possible
bias caused by clonal growth, care was taken to sample
from plants that were widely distant and to avoid
sampling from plants on the same host tree; this
reduced the scope for making larger samples in some
populations. Digital images were made using a Fuji
Finepix A303 digital camera while the leaves were still
fresh. After trimming off most of the petiole, the
leaves were attached with map pins to a backboard
made of rubberised carpet tiles with the abaxial side
exposed and photographed using a 15 cm measuring
rule as a scale. One source of variation in shape
capture arose from the fact that the leaves were not
strictly two-dimensional, but undulated to various
degrees especially along the margin. To reduce this
error, the pinning was carried out so that the leaf
undulation was evenly distributed around the margin
and the leaves were photographed directly overhead.
The images were prepared for outline digitisation
using the Gimp software package (GIMP 2006).
Monstera leaves are distichous and normally oblique,
i.e. one half of the leaf blade is wider than the other,
with successive leaves showing mirror-symmetry. Each
leaf was rotated so that the apex faced to the left with
the midrib more-or-less horizontal and ipped when
necessary to bring the broadest half of the blade onto
the lower side. The paint tool was used to adjust the
image to provide sufcient uniformity of tone along
the margin. Minor damage to the margin caused by
herbivores was also edited out.
The prepared images were then opened in tpsDig
(Rohlf 2004, ver. 1.40). The petiole-leaf blade junction
and leaf blade apex were digitised as landmarks 1 and
2 respectively. To remove ne scale noise along the
margin, the image was smoothed 8 to 10 times using
the Smooth image tool. The thresholded image was
viewed and checked using tpsDigs Image tools to
capture the outline as precisely as possible by optimis-
ing the threshold value. Editing-out of shadows was
sometimes necessary at this stage. After these steps,
the outline was digitised using the Outline object
tool. All the outlines of a population were thus written
into a single .tps le for further processing.
Size classication of leaves. A methodological compli-
cation was the conspicuous change in leaf shape and size
(heteroblasty) observed in Monstera adansonii during
development from juvenile to adult plants (Andrade &
195 COMPARATIVE MORPHOLOGY OF POPULATIONS OF MONSTERA ADANS.
The Board of Trustees of the Royal Botanic Gardens, Kew, 2008
Mayo 1998), which is even more pronounced in some
other species of the genus (Madison 1977; Ray 1983).
The total sample of leaves was therefore divided into
three size classes for analysis so that leaves at approxi-
mately the same heteroblastic stage would be compared.
In Monstera adansonii, this shape change is gradual
during the progression from the smallest to largest
leaves, but in M. praetermissa a sharp change in shape
was observed at about 15 cm length. Using this as a
guide, we selected 15 cm leaf blade length as the
boundary between juvenile and submature leaves. The
boundary between submature and mature was set at
30 cm leaf blade length after examination of 167 fertile
herbarium specimens at the Kew herbarium (K) of M.
adansonii var. klotzschiana and var. laniata, to establish an
approximate lower limit to the leaf blade size in mature
owering shoots.
Elliptic Fourier Analysis. Elliptic Fourier Analysis (EFA)
is a method for quantitatively characterising closed
outline shapes and has been used widely in many
biological elds, including taxonomy (e.g. Rohlf &
Archie 1984; Ferson et al. 1985; McLellan 1993; Cannon
& Manos 2001; Rumpunen & Bartish 2002; Jensen et al.
2002; Tatsuta et al. 2004; Yoshioka et al. 2004; Raveloson
et al. 2005). It is particularly useful for shapes which
possess few clearly homologous landmarks, as in the
present case, where only the petiole-leaf blade junction
and the leaf blade apex can be regarded as consistent,
biologically meaningful homologous sites. We used EFA
KBAS, LBAIlh,
LBAUb
KCEma, KCEpa
KFG, LFG
KCEub, PCEib, PCEub
KPA
KPER, LPET
KBAR, PBAR
KESli, KESst
KRJ
KSP
N
1000 km
PCEbat
Fig. 1. Geographical locations of Monstera populations studied in South America. Monstera adansonii var. klotzschiana. KBAR:
Bahia, Recncavo; KBAS: Bahia, Ubaitaba; KCEma: Cear, Serra de Maranguape; KCEpa: Cear, Serra de Aratanha, Pacatuba;
KCEub: Cear, Serra de Ibiapaba; KESli: Esprito Santo, Linhares; KESst: Esprito Santo, Santa Tereza; KFG: French Guiana; KPA:
Par, near Belm; KPER: Pernambuco, Recife; KRJ: Rio de Janeiro, near Rio de Janeiro city; KSP: So Paulo, Bertioga. Monstera
adansonii var. laniata. LPET: Pernambuco, Tapacur; LBAIlh: Bahia, Ilhus; LBAUb: Bahia, Ubaitaba; LFG: French Guiana. Monstera
praetermissa. PBAR: Bahia, Serra da Copioba; PCEbat: Cear, Serra de Baturit; PCEib: Cear, Serra de Ibiapaba (Ibiapina); PCEub:
Cear, Serra de Ibiapaba (Ubajara).
196 KEW BULLETIN VOL. 63(2)
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Table 1. Geographical details of sampled populations (20 populations, 1695 leaves) of Monstera adansonii var. klotzschiana, M. adansonii var. laniata and M. praetermissa.
Code and taxon Latitude longitude Population details
Country or
Brazilian state
Sample sizes (N)
Total
Mature
leaves
Submature
leaves
Juvenile
leaves
KBAR Monstera adansonii var. klotzschiana 125020S 391619W Recncavo (mean value for 2 localities) Bahia 43 22 14 7
KBAS " 141802S 391901W Ubaitaba Bahia 13 13 0 0
KCEma " 34818S 384246W Serra de Maranguape (mean value for
2 localities)
Cear 62 51 11 0
KCEpa " 35721S 383744W Serra de Aratanha, Pacatuba Cear 75 61 14 0
KCEub " 40119S 405143W Serra de Ibiapaba (Ibiapina, So Benedito,
Ubajara) (mean value for 3 localities)
Cear 179 21 90 68
KESli " 1993S 40415W Linhares Esprito Santo 14 6 8 0
KESst " 195757S 403158W Santa Tereza Esprito Santo 40 0 10 30
KFG " 43726N 523205W Petit Saut, Kaw, Matoury, Arata (mean
value for 4 localities)
French Guiana 189 127 24 38
KPA " 13032S 481759W Acar, Marituba Par 110 31 59 20
KPER " 8245S 344700W Recife (mean value for 2 localities) Pernambuco 61 26 35 0
KRJ " 225808S 431438W Rio de Janeiro city area (mean value for
2 localities)
Rio de Janeiro 114 0 63 51
KSP " 235114S 46820W Bertioga So Paulo 71 9 33 29
LBAIlh Monstera adansonii var. laniata 144619S 391319W Ilhus Bahia 21 6 15 0
LBAUb " 14182S 39191W Ubaitaba Bahia 44 14 20 10
LFG " 45432N 521626W Cayenne, Fort Diamant, Colline de
Montravel (mean value for 3 localities)
French Guiana 316 76 211 29
LPET " 800S 351058W Tapacur Pernambuco 37 18 19 0
PBAR Monstera praetermissa 124641S 390338W Serra da Copioba Bahia 8 8 0 0
PCEbat " 41633S 385726W Serra de Baturit Cear 39 0 28 11
PCEib " 35551S 405233W Serra de Ibiapaba (Ibiapina) Cear 171 53 78 40
PCEub " 34958S 405623W Serra de Ibiapaba (Ubajara) Cear 88 16 72 0
1
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to produce a matrix of 160 coefcients from which six
shape descriptor variables, the rst six principal com-
ponents, were derived.
The Fourier analysis was carried out using the
Morpheus et al. package (Slice 2008). Before running
the Elliptic Fourier Analysis module, the outlines were
standardised using the option Bookstein superimposi-
tion, which requires that the orientation, scale, star-
tingPt and location settings be switched off. The outlines
are then superimposed using two landmarks placed at
the base and apex of the leaf blade which equalises the
length and orientation of the interval between these
landmarks. In contrast, the default standardisation in
Morpheus involves equalising the length and orienta-
tion of the rst ellipse of all outlines. The justication
for using Bookstein superimposition is that the two
landmarks at either end of the midrib represent the
most consistent homologies recognisable in the leaves.
The Fourier analysis was carried out to a limit of 40
harmonics, resulting in a matrix of 164 elliptic Fourier
coefcients. The Fourier procedure reconstructs the
outlines to a level of approximation limited only by
the number of pixels used in the original digitisation,
so the decision on how many harmonics to set is to
some extent arbitrary (Rohlf & Archie 1984). After
comparing the reconstructed outlines by eye with the
originals we considered that with 40 harmonics, no
obvious artefactual distortions would be introduced
into the analyses. The rst 4 coefcients (harmonic 0)
are discarded because they correspond to an arbitrary
starting point of the outline tracing sequence rather
than any feature of the leaf shape, so the nal matrix
consisted of 160 variables (1
st
to 40
th
harmonic, each
harmonic consisting of 4 values).
Multivariate data analyses. Multivariate analyses of the
resulting coefcient matrices were carried out using
NTSYSpc ver. 2.20d (Rohlf 2005). Principal component
analysis of the coefcient matrices was used to reduce
the dimensionality of the data to a much smaller
number of uncorrelated shape descriptor variables. Each
of these variables was reied using the procedure
described by Yoshioka et al. (2004). First, the mean
vector of the Fourier coefcients was computed. Each
eigenvector was then multiplied by integer factors ( f )
from 1 to 5. The factored eigenvectors were then
separately added or subtracted from the mean vector.
The mean vector corresponds to the mean shape of the
data set. The effect of adding or subtracting a particular
factored eigenvector is to exaggerate or diminish its
effect on the mean shape while holding all the other
eigenvectors constant. The change in principal compo-
nent values corresponds to a change of f standard
deviations on the principal component concerned,
provided the principal component values have been
suitably scaled. The resulting vectors were then plotted
using the Fourplot module in NTSYSpc, which recon-
structs the outline from the set of Fourier coefcients
provided. These outlines visualise the shape changes
expressed independently by each of the shape variables
(principal components) we used. Figures 2, 3 and 4 show
this transformation from 2 to + 2 standard deviations.
We selected the rst six principal components (PCs)
to be treated as variables in a Canonical Variates Analysis
(CVA); together these accounted for 96.6% of the
variance in the original covariance matrix. The reduc-
tion to six PCs was necessary because once the leaf
samples had been divided into their size classes, some
population samples were reduced, in one case to only
8 individuals. The aim of the CVA, carried out using
NTSYSpc, was to show the maximum separation of the
centroids (multivariate means) of each population. The
resulting eigenvectors were used to project the matrix of
principal component scores for all the leaves analysed,
onto canonical variate axes. The matrix of pairwise
generalised distance between the population centroids
was used to generate Neighbor-Joining (NJ) trees, also
using NTSYSpc. As a comparison with the pattern based
on generalised distance, the mean vectors of the six
shape variables were computed for each population and
their pairwise Euclidean distances computed with
NTSYSpc. The resulting distance matrix was then used
to carry out a Principal Coordinates Analysis (PCoA),
incorporating a minimum spanning tree (MST).
To estimate intra-population variability, the Euclid-
ean distance between all pairs of individuals for each
population was computed, using the matrix of 6
principal component scores from the combined PCA
of all populations. Analyses of each size class were
carried out separately. The pairwise inter-individual
distances of each population were then summarised by
calculating the mean, standard deviation and coef-
cient of variation (Raveloson et al. 2005). The pairwise
differences in median Euclidean distance in the three
Cear populations were tested using the non-paramet-
ric Kruskal-Wallis test as implemented in PAST (Ham-
mer et al. 2001).
Similarity relationships between pairs of popula-
tions (based on the matrix of principal component
scores) were tested using multivariate procedures
implemented in PAST: NPMANOVA is a non-paramet-
ric analogue of MANOVA which calculates an F value
with signicance values computed by 5000 replicate
permutations of group membership (Table 2).
Results
The shape variables. In all three size classes, the rst
three principal components accounted for over 90%
of the total variance (Mature: 92.29%, Submature:
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1X
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- 1 s.d. - 2 s.d. Mean shape
+ 2 s.d. + 1 s.d.
Mature (54.67% of total variance)
Submature (71.28% of total variance)
Juvenile (62.85% of total variance)
Fig. 2. Reication of principal component 1 as shape variable Anisotropy. Derived from principal component analysis of elliptic
Fourier coefcient matrices of leaf outlines of Monstera adansonii var. klotzschiana, M. adansonii var. laniata and M. praetermissa.
Each outline is a 40-harmonic reconstruction from elliptic Fourier analysis data. Each row shows the trend of shape change along
principal component 1, in separate PCAs of mature (>=30 cm long), submature (15 to 29.9 cm long) and juvenile (< 15 cm long)
leaves. From left to right the outlines represent the principal component scores corresponding to: (mean 2 standard deviations),
(mean 1 s.d.), mean, (mean + 1 s.d.), (mean + 2 s.d.). Computed with NTSYSpc ver. 2.2.
199 COMPARATIVE MORPHOLOGY OF POPULATIONS OF MONSTERA ADANS.
The Board of Trustees of the Royal Botanic Gardens, Kew, 2008
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95.13%, Juvenile: 95.41%) and described distinct and
easily recognised trends of leaf outline shape change
which are illustrated in Figs. 2, 3 and 4. The fourth,
fth and sixth principal components expressed ob-
scure shape variations that were not easily interpreted
visually.
The rst principal component is named here the
Anisotropy shape variable (Fig. 2), because its most
conspicuous characteristic is the increase in length/
width ratio. This shape variable also expresses an
asymmetry at the leaf base which becomes more
pronounced in the transition from juvenile to mature
leaves. The second principal component, here termed
Dimidiate shape variable (Fig. 3) is characterised
mainly by the increasing difference in the width and
area of the two sides of the leaf blade, one side
eventually becoming very narrow. A secondary feature
is a slight basiscopic migration of the axis of widest
width. The third principal component, the Ovate
shape variable (Fig. 4), is characterised chiey by the
migration of the widest width axis giving a trend from
elliptic to ovate or vice versa (the orientation of the
principal component axes between analyses is arbi-
trary). A slight inverse dimidiate trend is also evident,
Table 2. Inter-population differences between nine populations of Monstera adansonii var. klotzschiana based on matrix of six
shape variables derived from elliptic Fourier analysis of mature leaf outlines.
KBAR KBAS KCEma KCEpa KCEub KFG KPA KPER KSP
KBAR
KBAS 0.009
KCEma 0.016 n.s.
KCEpa 0.000 n.s. 0.000
KCEub 0.048 0.027 0.003 0.000
KFG n.s. 0.011 0.011 0.000 0.009
KPA n.s. 0.027 0.011 0.000 n.s. n.s.
KPER n.s. 0.006 n.s. 0.000 0.002 0.020 0.042
KSP n.s. 0.002 0.000 0.000 n.s. 0.004 n.s. 0.001
Values shown are p-values (probability that two groups are the same) derived from Non-Parametric MANOVA implemented in PAST ver.
1.34 (Hammer et al. 2001); 5,000 replicates computed. See Table 1 for key to population codes.
0
0.001
0.002
0.003
0.004
0.005
0.006
0.007
0.008
PCEub PCEib LFG KCEpa KCEub KFG KPER LPET KCEma KPA LBAUb KBAR KSP
Populations
M
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mature submature juvenile
Fig. 5. Intra-population variability of leaf shape in Monstera adansonii var. klotzschiana, M. adansonii var. laniata and M.
praetermissa. For each population, the mean Euclidean distance between pairs of individuals is shown for three size classes (left bar:
mature leaves; middle bar: submature leaves; right bar [when data available]: juvenile leaves). Data is matrix of individual scores on
six shape variables (principal components) derived from EFA coefcients of leaf outlines. 95% condence limits shown. See Fig. 1
for key to population codes.
202 KEW BULLETIN VOL. 63(2)
The Board of Trustees of the Royal Botanic Gardens, Kew, 2008
i.e. the more ovate the leaf becomes, the less
dimidiate: this secondary trend is more distinct in
the submature and juvenile size classes.
Comparison of intrapopulational variability in Cear
populations of Monstera adansonii var. klotzschiana.
Juvenile leaves were only available for the Ubajara
population (KCEub), so comparisons of the three Cear
populations were made with submature and mature
leaves. Using mean Euclidean distance between pairs of
individuals as a measure of population variability,
submature leaves were more variable than mature leaves
(Fig. 5), with the largest discrepancy in size class
variability shown in the Pacatuba population (KCEpa).
Comparison of the mature leaves showed signicantly
greater variability in the Maranguape (KCEma) popu-
lation compared to the other two (Pacatuba: KCEpa;
Ubajara: KCEub), and non-signicant difference in the
variability of the Pacatuba and Ubajara populations.
Differences Between the Cear Populations of Monstera
adansonii var. klotzschiana. The three Cear populations
did not cluster together to form an exclusive group in
any analysis and the Pacatuba population (KCEpa)
was more distinct in a CVA of the three populations
(Fig. 6); the Maranguape and Ubajara populations
clustered closer together in both this and the NJ
analysis (Fig. 10). Pairwise tests between the three
populations (Table 2) showed signicant differences
between all populations for mature leaves. Submature
leaves were less distinct, with signicant differences in
all tests only between the Pacatuba and Ubajara
populations. Multivariate analyses (CVA and NJ)
showed somewhat different patterns of similarity
between the mature (Fig. 7) and submature (Fig. 8)
leaf classes; submature leaves of the Pacatuba popula-
tion were closer to var. laniata than to other var.
klotzschiana populations.
Comparison of the Cear Populations with those
of Monstera adansonii var. klotzschiana, var. laniata and
M. praetermissa in the Atlantic and Amazon Forests.
Comparison of intra-population variability, as estimated
by pairwise Euclidean distance between individuals,
showed that in most populations for which we had data
from all size classes, mature leaves are less variable than
submature ones (Fig. 5). In most cases, juvenile
populations of var. klotzschiana were less variable than
in the other two size classes.
Multivariate analysis (CVA, NJ) of the three size
classes produced quite similar results. Most populations
of var. klotzschiana grouped together, and populations
CV1
-0.0045 -0.0040 -0.0036 -0.0031 -0.0026
CV2
0.0027
0.0034
0.0041
0.0047
0.0054
Fig. 6. Canonical variates analysis of leaf outlines from populations of Monstera adansonii var. klotzschiana from Cear. Derived
from six shape variables extracted from elliptic Fourier coefcient data from mature leaves ( 30cm). Grey circles: Maranguape
(KCEma); white squares: Pacatuba (KCEpa); black diamonds: Ubajara (KCEub). Computed in NTSYSpc ver. 2.2.
203 COMPARATIVE MORPHOLOGY OF POPULATIONS OF MONSTERA ADANS.
The Board of Trustees of the Royal Botanic Gardens, Kew, 2008
CV1
-3.95 -2.56 -1.17 0.21 1.60
CV2
-2.50
-1.52
-0.55
0.42
1.40
KBAR
KBAS
KCEma
KCEpa
KCEub
KFG
KPA
KPER
LPET
KSP
LBAUb
LFG
PBAR
PCEib
PCEub
1X
-0.47
-0.22
0.02
0.27
0.52
5 2 1 4 7
1X
-0.47
-0.22
0.03
0.27
0.52
.23 .11 .02 .14 .26
-0.46
-0.22
0.03
0.28
0.
.30 .15 .00 .15 .30
1X
-0.46
-0.21
0.04
0.28
0.53
-0.28
-0.14 0.00 0.15 0.29
1X
-0.47
-0.22
0.02
0.27
0.51
.28 .14 .00 .15 .29
1X
-0.46
-0.22
0.03
0.28
0.53
9 5 0 5 0
1X
-0.47
-0.23
0.02
0.27
0.51
25 12 01 13 26
1X
-0.48
-0.23
0.02
0.27
0.52
0.26 0.13 0.00 0.14 0.27
1X
-0.48
-0.23
0.02
0.27
0.52
Y
-0.25
-0.12 0.01 0.13 0.26
1X
-0.48
-0.23
0.01
0.26
0.51
0.22 0.11 0.01 0.12 0.24
1X
-0.47
-0.22
0.02
0.27
0.51
22 10 02 14 26
-0.48
-0.24
0.01
0.26
0.50
26 13 00 14 27
1X
-0.48
-0.24
0.01
0.26
0.50
3 1 1 3 5
1X
-0.47
-0.22
0.02
0.27
0.51
0.25 0.13 0.00 0.13 0.26
1X
-0.48
-0.23
0.02
0.27
0.51
Y
-0.24
-0.12 0.01 0.13 0.26
CV1
-3.87 -1.94 -0.01 1.91 3.84
CV2
-2.88
-1.85
-0.82
0.21
1.25
PC1
PC2
PC3
PC4
PC5
PC6
Fig. 7. Canonical variates analysis of mature leaf outlines in nine populations of Monstera adansonii var. klotzschiana (KBAR, KBAS,
KCEma, KCEpa, KCEub, KFG, KPA, KPER, KSP), three populations of M. adansonii var. laniata (LBAUb, LFG, LPET) and three
populations of M. praetermissa (PBAR, PCEib, PCEub). Plot shows mean population shapes. CVA computed from six shape variables
(principal components) extracted by principal components analysis from a matrix of elliptic Fourier coefcients. Lower gure shows
vector diagram of eigenvectors in relation to the rst two canonical variates axes. Computed with NTSYSpc ver. 2.2.
204 KEW BULLETIN VOL. 63(2)
The Board of Trustees of the Royal Botanic Gardens, Kew, 2008
CV1
-1.95 1.03 2.51 4.00
CV2
-1.90
0.14
1.16
2.1
8
KBAR
KCEma
KCEpa
KCEub
KESli
KESst
KFG
KPA
KPER
LPET
KRJ
KSP
LBAIlh
LBAUb
LFG
PCEbat
PCEi b
PCEu b
1X
-6.82
-3.35
0.11
3.58
7.04
0 7 6 8 1
1X
-5.24
-2.53
0.18
2.89
5.60
60 3 4 1 8
1X
-6.44
-3.01
0.42
3.85
7.28
7 2 3 9 4
1
-43.10
-21.04
1.02
23.08
45.13
7 9 9 6 4
1
-0.48
-0.23
0.01
0.26
0.51
3 1 1 3 5
1X
-0.48
-0.24
0.01
0.26
0.50
0 0 1 1 2
1X
-0.47
-0.22
0.02
0.27
0.52
4 1 01 3 6
1X
-0.48
-0.24
0.01
0.26
0.51
4 1 1 3 5
1
-0.47
-0.22
0.03
0.27
0.52
23 1 01 4 26
1X
-0.49
-0.25
0.00
0.25
0.50
.11 .04 .02 .09 .16
1X
-0.48
-0.24
0.01
0.26
0.50
.16 .06 .03 .12 .21
1X
-0.48
-0.24
0.01
0.26
0.50
6 7 2 2 1
1
-0.49
-0.24
0.00
0.25
0.50
1
-0.49
-0.24
0.01
0.25
0.50
9 09 01 1 21
1
-0.49
-0.24
0.00
0.25
0.50
7 08 01 0 9
1X
-0.49
-0.24
0.01
0.25
0.50
0 9 1 2 2
-0.46
-0.22
0.03
0.27
0.52
9 4 1 6 1
1X
-0.46
-0.21
0.03
0.28
0.53
7 3 1 4 8
CV1
-1.95 -0.46 1.03 2.51 4.00
CV2
-1.90
-0.85
0.20
1.25
2.30
PC1
PC2
PC3
PC4
PC5
PC6
Fig. 8. Canonical variates analysis of submature leaf outlines in eleven populations of Monstera adansonii var. klotzschiana (KBAR,
KCEma, KCEpa, KCEub, KESli, KESst, KFG, KPA, KPER, KSP), four populations of M. adansonii var. laniata (LBAIlh, LBAUb, LFG,
LPET) and three populations of M. praetermissa (PCEbat, PCEib, PCEub). For further explanation see Fig. 7.
205 COMPARATIVE MORPHOLOGY OF POPULATIONS OF MONSTERA ADANS.
The Board of Trustees of the Royal Botanic Gardens, Kew, 2008
CV1
-3.80 -2.50 -1.19 0.11 1.41
CV2
-2.95
-1.54
-0.13
1.29
2.70
KBAR
KCEub
KESst
KFG
KPA
KRJ
KSP
LBAUb
LFG
PCEbat
PCEi b
1X
-0.49
-0.24
0.01
0.26
0.50
0.17
0.08
0.02
0.11
0.20
1X
-0.47
-0.22
0.02
0.27
0.51
23 11 01 12 24
1X
-0.49
-0.24
0.01
0.26
0.50
18 08 01 10 19
1X
-0.48
-0.23
0.01
0.26
0.50
22 11 01 13 25
1
-0.49
-0.24
0.01
0.25
0.50
13 06 01 08 16
1X
-0.48
-0.24
0.01
0.26
0.50
01X
-0.48
-0.24
0.01
0.26
0.50
1X
-0.47
-0.22
0.02
0.27
0.51
21 10 01 13 24
1X
-0.46
-0.21
0.04
0.29
0.53
27 13 01 15 29
1X
-0.49
-0.24
0.00
0.25
0.50
12 05 02 09 16
1X
-0.47
-0.23
0.02
0.26
0.51
18 08 02 12 22
CV1
-3.65 -2.12 -0.59 0.93 2.46
CV2
-2.87
-1.52
-0.17
1.18
2.53
PC1
PC2
PC3
PC4
PC5
PC6
Fig. 9. Canonical variates analysis of juvenile leaf outlines in seven populations of Monstera adansonii var. klotzschiana (KBAR,
KCEub, KESst, KFG, KPA, KRJ, KSP), two populations of M. adansonii var. laniata (LBAUb, LFG) and two populations of M.
praetermissa (PCEbat, PCEib). For further explanation see Fig. 7.
206 KEW BULLETIN VOL. 63(2)
The Board of Trustees of the Royal Botanic Gardens, Kew, 2008
of var. laniata and Monstera praetermissa formed sepa-
rate groups (Figs. 7, 8, 9 and 10). These taxonomically
correlated clusters were most distinct in mature leaves
(Fig. 7), with all populations of var. klotzschiana
forming a single cluster.
Pairwise NPMANOVA tests of mature-leaf popula-
tions of var. klotzschiana (Table 2) showed the most
signicant differences in pairs involving the Pacatuba
population (KCEpa). The results of these tests were
broadly consistent with a PCoA ordination of the
mean vectors of the six shape variable scores for each
of the nine var. klotzschiana populations (Fig. 11). The
latter plot emphasised the disparity between the three
brejo populations in Cear.
Mantel tests computed in NTSYSpc ver. 2.2 found
no signicant correlation between leaf outline shape
(measured as Euclidean distance between population
mean vectors of the six shape variables) and geo-
graphical distance between populations [10,000 ran-
dom permutations; p (random Z <= observed Z) =
0.6305, p (random Z >= observed Z) = 0.3695; matrix
correlation (normalised Mantel statistic Z): r = 0.06712]
or between leaf outline and genetic distance [data from
Andrade et al. 2007, genetic distance estimated as
pairwise F
ST
; 10,000 random permutations; p (random
Z <= observed Z) = 0.3308, p (random Z >= observed
Z) = 0.6692; matrix correlation (normalised Mantel
statistic Z): r = 0.1158].
Discussion
No strong geographical patterns emerged from the
analyses of var. klotzschiana populations. This contrasted
with the results obtained in our AFLP analysis (Andrade
et al. 2007), which showed a clear indication of greater
genetic diversity in Northeast Brazilian and Amazonian
populations and a signicant correlation of genetic and
geographical distance. The Cear brejo populations of
var. klotzschiana emerge from this study as distinct from
one another, at least using mature leaves. Although
there is considerable overlap (Fig. 5), their shapes are
signicantly different (Table 2). The Maranguape
population is more variable but the Pacatuba popula-
tion is the most distinct within Cear (Figs. 5, 8, 10).
The study shows that in var. klotzschiana, leaf outline
morphology expresses a degree of inter-population
Coefficient
0.00 0.65 1.29 1.94 2.58
KBAR
KPA
KSP
KBAS
KCEub
KCEma
KPER
KFG
KCEpa
LPET
LBAUb
LFG
PBAR
PCEib
PCEub
Fig. 10. Inter-population morphological distance of mature leaf outlines in nine populations of Monstera adansonii var.
klotzschiana (KBAR, KBAS, KCEma, KCEpa, KCEub, KFG, KPER, KSP), three populations of M. adansonii var. laniata (LBAUb, LFG,
LPET) and three populations of M. praetermissa (PBAR, PCEib, PCEub). Neighbor-Joining tree computed from a matrix of
generalised distance between population centroids derived from canonical variates analysis of six shape variables. Shape variables
(principal components) computed by principal components analysis from a matrix of elliptic Fourier coefcients. Computed with
NTSYSpc ver. 2.2.
207 COMPARATIVE MORPHOLOGY OF POPULATIONS OF MONSTERA ADANS.
The Board of Trustees of the Royal Botanic Gardens, Kew, 2008
divergence which is often signicant (Table 2). All the
most signicant inter-population differences involved
Cear populations, and the degree of morphological
divergence between the Cear brejo populations is
greater than between most population pairs from the
major forest blocks in Amazonia and the Atlantic
Forest). The Cear populations do not cluster togeth-
er as an exclusive group, but rather are mixed with
populations from Amazonia and the Atlantic forest
without an obvious pattern (Fig. 10). Broadly speak-
ing, the leaf outline data was successful as a marker
for the taxa (species and varieties) but showed no
clear geographical patterns at population level
within the best sampled taxon, var. klotzschiana. This
contrasts with our AFLP results (Andrade et al. 2007),
in which the molecular data revealed clear geograph-
ical structure.
This study demonstrates that leaf outline shape
alone is sufciently effective as a taxonomic marker to
cluster the populations of these three taxa into their
correct hierarchical taxonomic order successfully,
using mature leaves. Although population differences
were weaker, signicant divergence was observed. Leaf
outline shape, treated quantitatively can be a useful
tool for taxonomic comparisons down to population
level. This is somewhat surprising to anyone familiar
with the great variability of the leaves of Monstera
adansonii. Even more surprising is that the smaller leaf
size classes are almost as successful in discriminating
the taxa.
In var. klotzschiana the generally lower variability of
juvenile leaf shape, combined with its power to
discriminate the three taxa, is potentially of practical
value. In the damaged brejo forests, we observed that
where the forest had lost its continuous canopy
through human disturbance (e.g. So Benedito),
root-climbers like Monstera were represented almost
entirely by terrestrial and rupicolous juvenile forms,
because of the absence of suitable host trees. In this
situation, discrimination of juvenile forms of different
species is of practical importance in making invento-
ries of endangered biodiversity.
Clearly, a wider range of quantitative characters is
needed to capture the morphology more completely.
Leaf shape plays a signicant (but by no means
exclusive) role in the taxonomic denition of the taxa
we studied and therefore our nding that leaf outline
shape discriminates the taxa could be seen as a circular
argument. In our defence however, we would point out
that our eld identication of vegetative individuals
relied on a range of other characters including habitat,
stem morphology, leaf texture, leaf perforation pat-
terns and colour of the dried leaves. Furthermore this
study used quantitative estimates of variability and
difference in leaf shape and thus at the very least it
provides verication of grouping behaviour of a
morphological feature that is usually deployed intui-
tively in alpha taxonomy. Working from eld-collected
digital images proved to be a practical and effective way
to gather large samples from which quantitative data
could be captured semi-automatically. If extended to
other quantitative characters recoverable from images,
this approach could bring detailed and sophisticated
morphological studies at population level within the
reach of many workers, making it possible to work from
large samples without permanently damaging popula-
PCo1
-0.0026 -0.0014 -0.0003 0.0009 0.0021
PCo2
-0.0021
-0.0011
-0.0002
0.0008
0.0018
KBAR
KBAS
KCEma
KCEpa
KCEub
KFG
KPA
KPER
KSP
Fig. 11. Inter-population morphological distance of mature leaf outlines in nine populations of Monstera adansonii var.
klotzschiana (KBAR, KBAS, KCEma, KCEpa, KCEub, KFG, KPER, KSP). Principal coordinates analysis (PCoA) with minimum spanning
tree superimposed computed from a matrix of Euclidean distance between population mean vectors, derived from a matrix of six
shape variables. Shape variables (principal components) computed by principal components analysis from a matrix of elliptic Fourier
coefcients. Computed with NTSYSpc ver. 2.2.
208 KEW BULLETIN VOL. 63(2)
The Board of Trustees of the Royal Botanic Gardens, Kew, 2008
tions by removing signicant numbers of plants for
herbarium vouchering. This is increasingly a concern
for biologists who study surviving populations in small
and threatened habitats like the brejo forests of
Northeast Brazil.
Acknowledgements
We are most grateful to the following organisations and
persons for their essential help: FUNCAP (Fundaao
Cearense de Amparo Pesquisa), FAPESB (Fundao
de Amparo a Pesquisa do Estado da Bahia, WWF Brasil,
Kew Latin America Research Fellowships Programme
(Royal Botanic Gardens, Kew), Margaret Mee Fellow-
ships Programme (Fundao Botnica Margaret Mee,
Rio de Janeiro, Royal Botanic Gardens, Kew), Minis-
trio do Meio Ambiente (IBAMA, permit number 001/
2004, 042/04), Conselho Nacional de Desenvolvi-
mento Cientca e Tecnolgica (CNPq, MCT permit
number, 168/01), Dr Ana Maria Giulietti, Dr Robyn
Cowan, Anna Haigh, Dr Jean-Jacques de Granville and
the staff of the CAY herbarium in French Guiana,
Bertrand Goguillon and the staff of IRD (French
Guiana) for permission to collect, Dr Denis Barab,
Thales Alves Ribeiro, Sr. Jos Teodoro Soares, Rector
of the Universidade Estadual do Vale do Acara (UVA),
Sobral-CE.
Appendix
List of voucher specimens.
Voucher specimens are deposited in the following
herbaria: EAC = Universidade Federal do Cear,
Fortaleza, Cear, Brazil; HUEFS = Universidade Estadual
de Feira de Santana, Bahia, Brazil; K = Royal Botanic
Gardens, Kew, United Kingdom; UVA = Herbarium of
the Universidade Estadual Vale do Acara, Sobral, Cear.
Population codes are given in bold.
Monstera adansonii Schott var. klotzschiana (Schott)
Madison
KBAR: Brazil, Bahia, Serra da Copioba: I. M. Andrade
2771 (HUEFS), I. M. Andrade 2772 (HUEFS), I. M.
Andrade 2783 (HUEFS), I. M. Andrade 2784 (EAC),
I. M. Andrade 1996 (HUEFS).
KBAS: Brazil, Bahia, Ubaitaba: I. M. Andrade 2075
(HUEFS), I. M. Andrade 2077 (EAC), I. M. Andrade
2078 (HUEFS), I. M. Andrade 2079 (HUEFS).
KCEma: Brazil, Cear, Maranguape: I. M. Andrade 867
(UVA).
KCEpa: Brazil, Cear, Pacatuba: I. M. Andrade 685
(HUEFS), I. M. Andrade 706 (EAC), I. M. Andrade 707
(HUEFS).
KCEub: Brazil, Cear, So Benedito: I. M. Andrade 602
(EAC), I. M. Andrade 605 (EAC), I. M. Andrade 2613
(HUEFS). Ubajara: I. M. Andrade 2677 (HUEFS), I. M.
Andrade 2680 (HUEFS), I. M. Andrade 2709 (HUEFS),
I. M. Andrade 2722 (HUEFS).
KESli: Brazil, Esprito Santo, Linhares: I. M. Andrade
1618 (EAC), I. M. Andrade 1622 (EAC), I. M. Andrade
1628 (EAC).
KESst: Brazil, Esprito Santo, Santa Tereza: I. M. Andrade
764 (HUEFS), I. M. Andrade 767 (HUEFS).
KFG: French Guiana, Petit Saut: I. M. Andrade 2800
(EAC, HUEFS, K), I. M. Andrade 2802 (EAC, HUEFS, K).
KPA: Par, Acar: I. M. Andrade 2385 (HUEFS), I. M.
Andrade 2388 (HUEFS).
KPER: Brazil, Pernambuco, Recife: I. M. Andrade 1881
(HUEFS).
KRJ: Brazil, Rio de Janeiro, Vista Chinesa: I. M. Andrade
2222 (HUEFS), I. M. Andrade 2228 (HUEFS), I. M.
Andrade 2230 (HUEFS).
KSP: Brazil, So Paulo, Bertioga: I. M. Andrade 2121
(HUEFS), I. M. Andrade 2137 (HUEFS).
Monstera adansonii Schott var. laniata (Schott) Madison
LBAIlh: Brazil, Bahia, Ilhus, Centro de Pesquisas do
Cacau (CEPEC): I. M. Andrade 1983 (UVA).
LBAUb: Brazil, Bahia, Ubaitaba: I. M. Andrade 1962
(UVA).
LFG: French Guiana, Cayenne: I. M. Andrade 2815 (K).
LPET: Brazil, Pernambuco, Tapacur: I. M. Andrade
1933 (UVA).
Monstera praetermissa E. G. Gon. & Temponi
PBAR: Brazil, Bahia, Serra da Copioba: I. M. Andrade
2740 (UVA).
PCEbat: Brazil, Cear, Serra de Baturit, I. M. Andrade
s.n. (UVA).
PCEib: Brazil, Cear, Ibiapina, I. M. Andrade 2529 (UVA).
PCEub: Brazil, Cear, Ubajara, I. M. Andrade 2630 (UVA).
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