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New evidence on the origin of carnivorous plants

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COMMENTARY
New evidence on the origin of
carnivorous plants
Thomas J. Givnish1
Department of Botany, University of Wisconsin-Madison, Madison, WI 53706
Carnivorous plants have fascinated scientists
and the general public since the pioneering
studies of Charles Darwin (1). No doubt
part of their wide appeal is that carnivorous
plants have turned the evolutionary tables
on animals, consuming them as prey, with
the green predators often equipped with re-
markable lures, traps, stomachs, and—in
a few cases—extraordinary speed of move-
ment. To be considered carnivorous, a plant
must be able to absorb nutrients from dead
bodies adjacent to its surfaces, obtain some
advantage in growth or reproduction, and
have unequivocal adaptations for active
prey attraction, capture, and digestion (2,
3). Some carnivorous species [e.g., Pinguicula
(butterworts), Philcoxia] lack obvious attrac-
tants; some rely on passive pitfalls [e.g.,
Cephalotus (Australian pitcher plant), Sarra-
cenia (American pitcher plants)] rather than
active traps based on sticky tentacles [e.g.,
Byblis, Drosera (sundews)] or snap traps
[e.g., Dionaea (Venus fly-trap), Utricularia
(bladderworts)]; and some lack digestive
enzymes and instead depend on commensal
microbes or insect larvae to break down
prey (e.g., Brocchinia, Darlingtonia, some
species of Sarracenia). Based on these crite-
ria, today we recognize at least 583 species of
Table 1. Currently recognized groups of carnivorous plants
Order Family or clade
Poales Bromeliaceae I
Bromeliaceae II
Eriocaulaceae
Caryophyllales DNDD clade
Droseraceae
Nepenthaceae
Drosophyllaceae
Dioncophyllaceae
Oxalidales Cephalotaceae
Ericales RS-Actinidiaceae clade
Sarraceniaceae
Roridulaceae
Lamiales Byblidaceae
Lentibulariaceae
Plantaginaceae
Taxa include all members of each genus, except for the monocot genera in order Poales, where the number of
carnivorous species within the genus is listed. Independent origins of carnivory per se are indicated by boldface entries
in the family/clade column.
*Trap types indicated by superscript: L, lobster-pot trap; P, pitfall; S, snap trap; T, sticky trap.
www.pnas.org/cgi/doi/10.1073/pnas.1422278112
carnivorous plants in 20 genera, 12 families,
and 5 orders of flowering plants (Table 1).
Based on DNA sequence phylogenies, these
species represent at least nine independent
origins of the carnivorous habit per se, and
at least six independent origins of pitfall
traps, five of sticky traps, two of snap traps,
and one of lobster-pot traps. To the extent
to which molecular phylogenies have been
calibrated against the ages of fossils of other
plants, these origins of carnivory appear to
have occurred between roughly 8 and 72
million years ago (Mya). In PNAS, Sadowski
et al. (4) contribute to our understanding
of the origins of plant carnivory by describ-
ing the first fossilized trap of a carnivorous
plant, a fragment of a tentacled leaf pre-
served in Baltic amber from 35 to 47
Mya, and allied to modern-day Roridula of
monogeneric Roridulaceae (Ericales) from
South Africa.
As with most carnivorous plants, the two
living species of Roridula today grow on
open, extremely infertile, moist sites. The
occurrence of carnivorous plants on nutrient-
poor substrates has been understood since
Darwin showed that such plants augment
their supply of mineral nutrients through
prey capture. The restriction of carnivorous
Genus/genera* No. of taxa
P
Brocchinia
CatopsisP
PaepalanthusP
AldrovandaS, DionaeaS, DroseraT
NepenthesP
DrosophyllumT
TriphyophyllumT
CephalotusP
DarlingtoniaP, HeliamphoraP, SarraceniaP
RoridulaT
ByblisT
GenliseaL, PinguiculaT, UtriculariaS
PhilcoxiaT
COMMENTARY
plants to open, infertile, moist sites, however,
remained unexplained until modern cost-
benefit models showed that carnivores are
likely to obtain an advantage in growth rela-
tive to noncarnivores only on such sites,
where nutrients and nutrients alone limit plant
growth, and where carnivory can accelerate
photosynthesis and the conversion of photo-
synthate to new leaf tissue while decreasing
allocation to root tissue (2, 3, 5, 6). Wet soils
and fire can favor carnivorous plants, by mak-
ing N more limiting for growth while making
light and water less limiting (3). The wet,
sandy, fireswept sites in fynbos occupied by
Roridula (6) should thus favor carnivory,
and indeed Roridula often grows in association
with large numbers of carnivorous sundews.
Roridula, however, is in other respects
highly unusual as a carnivorous plant. Al-
though its glistening, glandular tentacles
do trap large numbers of insects, the secre-
tions are resinous rather than aqueous, and
so cannot support the activities of digestive
enzymes. It does not secrete proteolytic en-
zymes; several authors thus argued that
Roridula could not be carnivorous because
it could not digest prey or absorb the min-
erals released (7, 8). The resinous nature of
Roridula secretions may be an adaptation to
the summer drought in the Mediterranean
climate it now occupies, in that they do not
lose volume or stickiness during long periods
of drought; the secretions also do not dissolve
during winter rains (9). It turns out that cer-
tain hemipterans (Pameridea) are capable of
negotiating the glandular leaves of Roridula
without becoming entangled; they eat the
2
prey immobilized by the plant, and then
1
N from their excretions is absorbed by
1
Roridula (Fig. 1). This process substantially
115 augments the N supply to the plants, with
90 the plants obtaining 70% or more of their
1 nitrogen supply in this fashion (7, 10). The
1 mutualism appears stabilized by nonlinear
1 interactions: excess densities of Pameridea
turn counterproductive as the bugs switch
32 to sap-sucking in the absence of prey, leading
2 to negative impacts on Roridula and, ulti-
6 mately, on the bugs themselves (11).
330
1
Author contributions: T.J.G. wrote the paper.
The author declares no conflict of interest.
See companion article 10.1073/pnas.1414777111.
1
Email: givnish@wisc.edu.
PNAS Early Edition | 1 of 2

Fig. 1. Growth form of Roridula gorgonias at Fernkloof Nature Reserve near Hermanus, showing glandular ten-
tacles that immobilize insect prey. Close-up of leaves, showing a Pameridea bug (center) that eats immobilized prey
and delivers nutrients to the plant via excreta.
Although the Roridula system is truly re-
markable, similar kinds of complex digestive
mutualisms may occur in other carnivorous
plants. For example, Nepenthes bicalcarata
provides domatia for ants, despite ants being
the most frequent prey of many Nepenthes.
Givnish (5) and Hölldobler and Wilson (12)
proposed that the resident ants and plants
might have a mutualistic relationship of some
kind. In fact, the resident ant Camponotus
schmitzi protects N. bicalcarata from weevils
that attack their tendrils, and in addition
facilitates the plant’s uptake of nutrients
(13). The ants can swim in the pitcher fluid
without adverse effect, retrieve large prey
items, and excrete wastes into the pitcher,
accelerating nutrient uptake; ant wastes ac-
count for 42–76% of total N uptake and ants
prolong pitcher lifetimes (13). In other sys-
tems, the prey processed by a digestive mu-
tualist may not even be captured by the
plant’s own traps. Nepenthes lowii attracts
tree shrews (Tupaia montana) to their excep-
tionally large, broad traps with secreted
rewards, and they defecate into the pitcher
while marking it as their territory. Their feces
account for 57–100% of all leaf N (14).
Nepenthes rafflesiana var. elongata, with
smaller but elongate traps, provides a roost
for a small bat and obtains nutrients from
its feces (15). Whether these systems are best
viewed as coprophagy or indirect forms of
carnivory involving digestive mutualists that
deliver the remains of prey is worth debating.
Clearly, however, both plants benefit from
animals whose death results in their acquisi-
tion of nutrients; we might consider them
2 of 2 | www.pnas.org/cgi/doi/10.1073/pnas.1422278112
“apparent carnivorous plants,” in homage to
Holt’s concept of apparent competition (16).
The new fossil Roridula not only is the first
fossil trap leaf uncovered, it is one of the very
few undoubted fossils of carnivorous plants
of any kind. Archaeamphora from Chinese
sediments 112 Mya was originally described
as Sarraceniaceae, but now there is strong
doubt that it was a member of that family
or even a carnivorous plant; the unusual
leaves may simply not have been traps (17).
Paleoaldrovanda, putatively a member of Dro-
seraceae based on a “seed,” may actually have
1 Darwin C (1875) Insectivorous Plants (Appleton and Co., London).
2 Givnish TJ, Burkhardt EL, Happel RE, Weintraub JW (1984)
Carnivory in the bromeliad Brocchinia reducta, with a cost/benefit
model for the general restriction of carnivorous plants to sunny,
moist, nutrient-poor habitats. Am Nat 124(4):479–497.
3 Ellison M, Adamec L (2011) Ecophysiological traits of terrestrial
and aquatic carnivorous plants: Are the costs and benefits the same?
Oikos 120(11):1721–1731.
4 Sadowski E-M, et al. (2014) Carnivorous leaves from Baltic amber.
Proc Natl Acad Sci USA, 10.1073/pnas.1414777111.
5 Givnish TJ (1989) Ecology and evolution of carnivorous plants.
Plant–Animal Interactions, ed Abrahamson WG (McGraw-Hill, New
York), pp 243–290.
6 Ellison AM, Gotelli NJ (2009) Energetics and the evolution of
carnivorous plants—Darwin’s ‘most wonderful plants in the world’
J Exp Bot 60(1):19–42.
7 Ellis AG, Midgley JJ (1996) A new plant-animal mutualism involving
a plant with sticky leaves and a resident hemipteran. Oecologia
106(4):478–481.
8 Juniper BE, Robias RJ, Joel DM (1989) The Carnivorous Plants
(McGraw-Hill, New York).
9 Voigt D, Gorb S (2010) Desiccation resistance of adhesive
secretion in the protocarnivorous plant Roridula gorgonias as an
adaptation to periodically dry environment. Planta 232(6):
1511–1515.
10 Anderson B, Midgley JJ (2003) Digestive mutualism, an alternative
pathway in plant carnivory. Oikos 102(1):221–223.
been a fossil insect egg (18). The remaining
fossils considered legitimate remains of carniv-
orous plants include one seed (now destroyed)
of Byblis (Byblidaceae) from Australia (19),
and palynomorphs possibly allied with
Nepenthaceae (20). The last two fragments,
however, do not demonstrate that the plants
to which they belonged were, in fact, carniv-
orous, which makes the find by Sadowski et al.
(4) particularly important. The age of the am-
ber Roridula, 35–47 Mya, nicely brackets the
divergence between Roridula and noncarnivo-
rous Actinidiaceae roughly 39 Mya, as esti-
mated from a calibrated DNA phylogeny
(21). This result lends credence to the age
estimates based on molecular data, and to
the inference from phylogenetic reconstruc-
tion that early Roridulaceae were carnivorous.
The identity of the fossil Roridula appears to
be beyond doubt. The former occurrence of
Roridula around the Baltic—whereas its pres-
ent-day distribution is restricted to the Cape
Floristic Province of southwest South Africa—
implies that this group was once far more
widespread. The distributions of families in
the Clethraceae-Sarraceniaceae-Roridulaceae-
Actinidiaceae clade suggest that it originated
in southeastern North America or northern
South America. In the next few years, further
investigations of the Baltic amber might tell us
what other plants grew in association with
fossil Roridula, and thus the nature of the
vegetation in which fossil Roridula grew.
Based on cost-benefit models, the distribution
of present-day Roridula, and the current dis-
tributions of almost all other carnivorous
plants, it seems most unlikely that fossil
Roridula grew below a dense canopy of the
conifer forests that produced amber!
11 Anderson B, Midgley JJ (2007) Density-dependent
outcomes in a digestive mutualism between carnivorous
Roridula plants and their associated hemipterans. Oecologia
152(1):115–120.
12 Hölldobler B, Wilson EO (1990) The Ants (Springer, Berlin).
13 Bazile V, Moran JA, Le Moguédec G, Marshall DJ, Gaume L
(2012) A carnivorous plant fed by its ant symbiont: A unique multi-
faceted nutritional mutualism. PLoS ONE 7(5):e36179.
14 Clarke CM, et al. (2009) Tree shrew lavatories: A novel nitrogen
sequestration strategy in a tropical pitcher plant. Biol Lett 5(5):
632–635.
15 Grafe TU, Schöner CR, Kerth G, Junaidi A, Schöner MG (2011) A
novel resource-service mutualism between bats and pitcher plants.
Biol Lett 7(3):436–439.
16 Holt RD (1977) Predation, apparent competition, and the
structure of prey communities. Theor Popul Biol 12(2):197–229.
17 Brittnacher J (2013) Phylogeny and biogeography of the
Sarraceniaceae. Carniv Plant Newsletter 42(3):99–106.
18 Hermanová Z, Kvaček J (2010) Late Cretaceous
Palaeoaldrovanda, not seeds of a carnivorous plant but eggs of an
insect. J Natur Hist Mus (Prague) 179(9):105–118.
19 Conran J, Christophel D (2004) A fossil Byblidaceae seed from
Eocene South Australia. Int J Plant Sci 165(4):691–694.
20 Kumar M (1995) Pollen tetrads from Palaeocene sediments of
Meghalaya, India: Comments on their morphology, botanical affinity
and geological records. Palaeobot 43(1):68–81.
21 Ellison AM, et al. (2012) Phylogeny and biogeography of
the carnivorous plant family Sarraceniaceae. PLoS ONE 7(6):e39291.
Givnish
Carnivorous leaves from Baltic amber
Eva-Maria Sadowskia, Leyla J. Seyfullaha, Friederike Sadowskib, Andreas Fleischmannc, Hermann Behlingd,
and Alexander R. Schmidta,1
a
Department of Geobiology, University of Göttingen, 37077 Göttingen, Germany; bInstitute for Interdisciplinary Research on Conflict and Violence, Bielefeld
University, 33615 Bielefeld, Germany; cBotanische Staatssammlung München, 80638 Munich, Germany; and dDepartment of Palynology and Climate
Dynamics, Albrecht von Haller Institute of Plant Sciences, University of Göttingen, 37073 Göttingen, Germany
Edited by Peter R. Crane, Yale School of Forestry and Environmental Studies, New Haven, CT, and approved November 3, 2014 (received for review August
1, 2014)
The fossil record of carnivorous plants is very scarce and macro-
fossil evidence has been restricted to seeds of the extant aquatic
genus Aldrovanda of the Droseraceae family. No case of carnivo-
rous plant traps has so far been reported from the fossil record.
Here, we present two angiosperm leaves enclosed in a piece of
Eocene Baltic amber that share relevant morphological features
with extant Roridulaceae, a carnivorous plant family that is today
endemic to the Cape flora of South Africa. Modern Roridula
species are unique among carnivorous plants as they digest prey
in a complex mutualistic association in which the prey-derived
nutrient uptake depends on heteropteran insects. As in extant
Roridula, the fossil leaves possess two types of plant trichomes, in-
cluding unicellular hairs and five size classes of multicellular stalked
glands (or tentacles) with an apical pore. The apices of the narrow
and perfectly tapered fossil leaves end in a single tentacle, as in both
modern Roridula species. The glandular hairs of the fossils are re-
stricted to the leaf margins and to the abaxial lamina, as in extant
Roridula gorgonias. Our discovery supports current molecular age
estimates for Roridulaceae and suggests a wide Eocene distribution
of roridulid plants.
plant carnivory | Roridulaceae | Eocene | Ericales
P lant carnivory is traditionally defined as the attraction, cap-
ture, and digestion of prey by vegetative traps, with the
subsequent uptake of nutrients (1, 2). Some carnivorous plants,
however, challenge the boundary of the botanical carnivory con-
cept because they depend on commensal organisms for the di-
gestion of their prey (2, 3). The most famous representative
of those plants is Roridula, placed in the monogeneric family
Roridulaceae that is endemic to a few localities in the south-
western Cape of South Africa (4, 5).
The resinous glandular leaves of both extant species, Roridula
dentata and Roridula gorgonias, capture plenty of arthropods.
The sticky trapping glue of Roridula is a viscous lipophilic resin
containing triterpenoids as major component, which does not
allow dissolution of digestive enzymes (6). Consequently, the
secretory glands of Roridulaceae lack enzymatic activity (7, 8).
For prey-derived nutrient uptake, Roridula depends on two ob-
ligately associated heteropteran Pameridea species (family Miridae,
“capsid bugs”), which feed on the trapped animals (5, 9). In this
“digestive mutualism” (10), the nutrient-rich fecal compounds of
these “Roridula bugs” are incorporated by Roridula through
nanometer-sized cuticular gaps and serve for a better alimenta-
tion in a nutrient-poor habitat (7, 8, 10, 11). The benefit of nu-
trient uptake from captured prey is the essential criterion
for the concept of botanical carnivory (1, 2) and thus includes
Roridulaceae (11, 12).
Here, we report two leaf fossils from Eocene Baltic amber
possessing the relevant morphological features of an adhesive
flypaper trap plant that we assign to the Roridulaceae lineage
(Figs. 1–3). Both specimens originate from the Jantarny amber
mine near Kaliningrad (Russia). The amber-bearing sediments
of this fossil site date to 35–47 million years ago (13, 14).
www.pnas.org/cgi/doi/10.1073/pnas.1414777111
Results
The linear-lanceolate leaves are 5 and 4.5 mm long and 0.2 mm
wide at the base, and they narrow gradually toward the leaf tip,
which terminates in a stalked gland (tentacle; Fig. 1). The leaves
possess two trichome types: tentacles and nonglandular hyaline
hairs (Figs. 1 and 2). The hyaline trichomes are located on both
sides and the margins of the lamina, whereas the tentacles are
exclusively found along the margins and on the abaxial side
without a definite arrangement (Fig. 1). The tentacles are multi-
cellular, consisting of a tapering stalk and a clavate to ovoid
glandular head, which shows a small pore at the center of its distal
side (Fig. 1 E and F). The stalks of the glands measure between 20
and 350 μm in length (Figs. 1 and 2 A and C), whereas an ex-
EVOLUTION
ceptional stalk exceeds this size, reaching 1.4 mm (Fig. 3A). As
with the stalks, the glandular heads vary in size (20–120 μm long,
10–40 μm wide). Adhered organic remains as well as trichomes of
other plants attached to the glandular tentacle heads (Figs. 1
A and D and 3A) indicate that they excreted a sticky secretion, as
known from adhesive traps of extant carnivorous plants. The non-
glandular trichomes are hyaline, unicellular, and arcuate to straight,
tapering toward an acute apex (Figs. 1C and 2E). Their length
ranges from 10 to 80 μm, and their width reaches up to 12 μm.
Both leaves exhibit a well-preserved epidermis with small tetrag-
onal cells at the leaf base and elongated larger cells from the middle
part toward the leaf tip. These cells measure 3–54 × 6–18 μm.
Stomata of 20–38 × 15–25 μm are present on the abaxial leaf side
(Figs. 1C and 2G).
Our statistical cluster analyses (SI Text) revealed that the
fossil specimens and Roridula gorgonias show the same mor-
phological pattern among the tentacles. R. gorgonias exhibits five
Significance
Amber, fossil tree resin, preserves organisms in microscopic
fidelity, and frequently fossils preserved in amber are other-
wise absent in the entire fossil record. Plant remains, however,
are rarely entrapped in amber, compared with the vast amount
of insects and other animals. Our newly discovered fossils from
Eocene Baltic amber are the only documented case of fossilized
carnivorous plant traps and represent the first fossil evidence
of the carnivorous plant family Roridulaceae, which is today a
narrow endemic of South Africa. Hence, our results shed light
onto the paleobiogeography of the Roridulaceae, indicating a
wide Eocene distribution of the roridulid ancestors and chal-
lenging previous notions about a Gondwanan origin of this
plant family.
Author contributions: E.-M.S., L.J.S., H.B., and A.R.S. designed research; E.-M.S., L.J.S., F.S.,
A.F., H.B., and A.R.S. performed research; E.-M.S., L.J.S., F.S., A.F., and A.R.S. analyzed
data; and E.-M.S., L.J.S., A.F., and A.R.S. wrote the paper.
The authors declare no conflict of interest.
This article is a PNAS Direct Submission.
1
To whom correspondence should be addressed. Email: alexander.schmidt@geo.uni-
goettingen.de.
This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.
1073/pnas.1414777111/-/DCSupplemental.
PNAS Early Edition | 1 of 6
Fig. 1. Carnivorous leaves from Eocene Baltic amber. (A) Overview of the leaf enclosed in amber specimen GZG.BST.27310 showing the adaxial tentacle-free
side in slightly oblique view and stalked glands at the margin and on the abaxial side; arrowhead points to the exceptional long tentacle stalk with several
branched oak trichomes attached. (B) Overview of the leaf enclosed in amber specimen GZG.BST.27311, showing abundant tentacles on the abaxial side. (C)
Margin of abaxial leaf surface with tentacles of different size classes and nonglandular hyaline trichomes. (D) Leaf apex tapering into a sole tentacle. (E and F)
Glandular heads with central pore (arrowheads) from both leaves. (Scale bars: A and B, 1 mm; C and D, 100 μm; E, 10 μm; F, 40 μm.)

2 of 6 | www.pnas.org/cgi/doi/10.1073/pnas.1414777111 Sadowski et al.

 EVOLUTION

Fig. 2. Morphological comparison of the carnivorous leaf fossils from Baltic amber (Left) and extant Roridula species (Right). (A and B) Leaf tip ending in a sole
tentacle. (C and D) Stalked glands of different size classes. (E and F) Hyaline unicellular nonglandular trichomes. (G and H) Epidermal cells and stomata. (I–L) Mul-
ticellular tentacles. (A, C, E, and G) GZG.BST.27310. (I and J) GZG.BST.27311. (B, D, K, and L) R. gorgonias. (F and H) R. dentata. (Scale bars: A–D, 100 μm; E–L, 50 μm.)

Sadowski et al. PNAS Early Edition | 3 of 6

Fig. 3. Carnivorous leaf from Eocene Baltic amber (A and B; GZG.BST.27310) and leaves of extant Roridula gorgonias (C and D). (A) Exceptionally
long tentacle stalk (with several branched oak trichomes attached) of the fossil leaf representing the fifth size class of stalked glands. (B and C ) Overviews
showing the tentacle-free adaxial surface and tentacles along the leaf margins. (D) Partial leaf tip showing different size classes of stalked glands. (Scale
bars: A, 100 μm; B, 500 μm; C and D, 1 mm.)
size clusters of tentacle stalk lengths whereas four clusters were
detected for the amber inclusions. As an outlier, the longest ten-
tacle was excluded from the cluster analyses of the fossils, but its
presence indicates that a fifth size class was present in the Eocene
leaves, as in extant Roridulaceae (Fig. 4).
Discussion
Although glandular secreting trichomes appear in about 30% of
vascular plants (15), the unique character combination in the
fossils bears most similarities to extant representatives of the
Roridulaceae. They share the long, narrow, and perfectly ta-
pered leaf lamina ending in a single tentacle, the presence and
morphology of two trichome types (tentacles and nonglandular
hairs), the possession of glandular hairs along the leaf margins
and on the abaxial lamina, the tentacle head with a central pore,
and the size and shape of the epidermal cells and stomata. The
glabrous adaxial side of the amber inclusions and the hyaline
trichomes being located on both leaf surfaces only appear in the
sepals of extant Roridula gorgonias. Besides smaller tentacle
sizes, the fossils are distinguished from leaves of extant Roridula
species by the absence of a prominent midrib on the abaxial leaf
side, and are thus most similar to sepals of Roridula gorgonias.
4 of 6 | www.pnas.org/cgi/doi/10.1073/pnas.1414777111
Extant Roridula plants are very effective traps for all kinds of
arthropods due to the sticky resinous trapping glue and the hier-
archical organization of the tentacles into functional units for ef-
fective prey capture (16, 17). The longest tentacles make the first
contact with the prey. Due to the high flexibility of these prom-
inent tentacles, the moving prey then gets stuck to the medium-
sized tentacles, which slow down the caught animal. Finally, the
smallest and stiffest tentacles immobilize the prey (16). As in
modern Roridulaceae, the leaf fossils have different size classes of
tentacles that fulfill the functional roles for prey capture (entan-
glement, slow-down, and immobilization) and comply with the
requirements for a carnivorous nature. In addition, the pore of the
tentacle heads distinguishes the fossils from any other extant
carnivorous plants with glandular adhesive traps such as sundews
(Drosera) (3, 18, 19).
In the fossil record, evidence of carnivorous plants is exceed-
ingly rare and macrofossils are restricted to seeds of the aquatic
carnivore Aldrovanda (Droseraceae), which are recorded since
the Eocene (20, 21). Hence, the fossil leaves from Baltic amber
are (to our knowledge) the first documented case of carnivorous
plant traps being fossilized.
Sadowski et al.
 A 1500
Amber fossils
1200
stalk length (µm)
900
600
300
0 amber piece that derives from Jantarny mine near Kaliningrad (Russia).
Size class 1 Size class 2 Size class 3 Size class 4 Size class 5
(n=43) (n=28) (n=18) (n=8) (n=1)
3500
B Roridula gorgonias
2800
stalk length (µm)
2100
1400
700
0 Leaves of extant Roridula gorgonias and R. dentata (Roridulaceae) were
Size class 1 Size class 2 Size class 3 Size class 4 Size class 5
(n=61) (n=18) (n=3) (n=9) (n=5)
Fig. 4. Tentacle size classes of the fossil leaves and extant Roridula
gorgonias based on the results of the cluster analyses and the tentacle
stalk length. (A) Size classes of the fossil leaves, including the outlier
which we interpret to represent size class 5. (B) Size classes of Roridula
gorgonias. n indicates the number of tentacles per size class.
The occurrence of Eocene Roridulaceae is consistent with
recent divergence time estimates for a split of Sarraceniaceae
(carnivorous American pitcher plants) from the Roridulaceae–
Actinidiaceae clade about 48.6 million years ago, whereas the
most recent common ancestor of Roridulaceae and Actinidia-
ceae was estimated at 38.1 million years ago (22). The age of
these Ericales lineages is further supported by Late Cretaceous
fossil flowers with affinities to the Actinidiaceae and Clethraceae
families (23). The sediments containing the majority of Baltic
amber are 35–47 million years old (13, 14). Thus, the amber
fossils probably represent an early representative of the
Roridulaceae lineage.
The geologic setting of the Baltic amber deposit and the
paleobotanical record suggest that coastal areas with carbonate-
free, nutrient-poor soils and swamp depressions harbored well-
structured mixed forests of angiosperm and conifer trees with
intermixed open habitats growing in a subtropical to warm-
temperate climate (24–27).
The presence of Eocene roridulid plants in the northern
hemisphere challenges notions about the biogeographical history
of extant Roridulaceae, which were previously assumed to rep-
resent “old Cape elements,” paleoendemics of Gondwanan ori-
gin, dating back to up to 90 million years (28, 29). Thus, the leaf
fossils represent an example of pseudo-Gondwanan relicts, extinct
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in Europe today and restricted to particular areas of the southern
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Materials and Methods
Provenance of the Amber Piece. The leaf inclusions were discovered in an
Amber in this locality is mined in the “Blue Earth” layer, which is Priabonian
in age (late Eocene, 35 million years minimum age) (13, 14). The amber piece
was obtained from the collection of Christel and Hans Werner Hoffeins
(Hamburg, Germany).
Microscopy and Imaging. The original 39 × 21 × 5-mm piece of amber was
ground and polished manually with wet silicon carbide papers (grit from
25.8- to 5-μm particle size; firm Struers). Two amber fragments measuring
21 × 14 × 3 and 24 × 19 × 3 mm with one leaf inclusion each were obtained
by cutting the amber piece with a dental drill. The amber pieces are housed
in the Geoscientific Collections of the Georg August University Göttingen
(Göttingen, Germany) (collection numbers GZG.BST.27310 and GZG.BST.27311).
obtained from cultured specimens of Thomas Carow (Nüdlingen, Germany)
EVOLUTION
and A.R.S. The leaf inclusions and the extant Roridulaceae plant material
were examined under a Carl Zeiss Stemi 2000 dissection microscope and a
Carl Zeiss AxioScope A1 compound microscope, each equipped with a Canon
60D digital camera. In most instances, incident and transmitted light were
used simultaneously. The images of Figs. 1 A–D, 2, and 3 A and B are digitally
stacked photomicrographic composites of up to 130 individual focal planes
obtained using the software package HeliconFocus 5.0 for a better illustra-
tion of the 3D structures.
Statistics. According to Voigt et al. (16), Roridula gorgonias possesses three
tentacle size classes that allow a very effective capture of prey. To test
whether the tentacle size classes are present in the amber specimens, the
tentacle measurements of the amber specimens, Roridula gorgonias and
R. dentata were statistically evaluated, applying hierarchical cluster analyses
with the statistics package IBM SPSS 21. In total, measurement values of 103
tentacles from both leaf inclusions and 103 tentacles from each extant
Roridula species were used, comprising the stalk length, the stalk base and
tip width, the gland length, and the gland width. The hierarchical cluster
analyses were computed using Ward’s method. The resulting clusters were
optimized with the nonhierarchical k-means method and tested statistically,
using three criteria suggested by Bacher (30), which are η-squared (η2), F-max,
and the proportional reduction of error (PRE). The best cluster solution is
the one where the values of η2 and PRE do not show any considerable im-
provement in the subsequent solution. Furthermore, the F value and η2 should
be maximal, whereas PRE is supposed to be low. The number of clusters also
should be selected with regard to the content and the underlying theoretical
model (30). Results of the statistical analyses are provided in SI Text.
ACKNOWLEDGMENTS. We thank Christel and Hans Werner Hoffeins
(Hamburg) for providing the amber specimen and Thomas Carow (Nüdlingen)
for providing a plant of Roridula dentata for study. We are grateful to Julia
Gundlach (Bielefeld University), Dorothea Hause-Reitner, and Gerhard
Hundertmark (University of Göttingen) for assistance, and to two anonymous
reviewers for constructive suggestions.
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