Arthropod-Plant Interactions (2010) 4:69–79

DOI 10.1007/s11829-010-9088-1

ORIGINAL PAPER

Locomotion in a sticky terrain

Dagmar Voigt • Stanislav Gorb

Received: 6 October 2009 / Accepted: 2 February 2010 / Published online: 18 February 2010
Ó The Author(s) 2010. This article is published with open access at Springerlink.com

Abstract The mirid bug Pameridea roridulae lives mu- mirid bug species from the tribus Dicyphini: (1) avoidance
talistically on the protocarnivorous plant Roridula gorgo- strategy, characterised by the slim body held at a large
nias. The latter resembles an effective, three-dimensional distance from the plant surface by using long, slender legs,
flypaper trap which captures numerous flying insects. We and (2) defense strategy, where trapping of the heavy bugs,
have recently shown that P. roridulae bugs are not trapped situated close to the plant surface, is overcome by gener-
by the plant, because they are covered with a layer of ating strong forces during locomotion and by having a
epicuticular grease, which is considerably thicker than in thick anti-adhesive epicuticular greasy layer on the bugs’
other insects. The present study demonstrates that the bugs’ cuticle.
morphology and locomotory characteristics also contribute
to their specialisation for life on the adhesive plant surface. Keywords Attachment  Biomechanics  Bryocorinae 
A structural analysis of the mirid bug’s attachment system, Heteroptera  Insect-plant interactions  Miridae 
and an experimental study on its attachment ability were Traction force  Trichomes
carried out. In traction force tests, maximum forces of
8.8 mN were measured on adaxial R. gorgonias leaves,
corresponding to 126 times the bug’s body weight. On Introduction
smooth surfaces, generated forces were only 47 times the
bug’s body weight. Compared to closely related mirid bug South African bugs Pameridea roridulae Reuter (Het-
species avoiding contact with plant adhesive secretion, eroptera, Miridae, Bryocorinae, Dicyphini) are obligately
P. roridulae is distinctly stronger and heavier, and holds its associated with protocarnivorous plants Roridula gorgo-
body close to the plant substrate. Two locomotion strate- nias Planch. (Roridulaceae; Reuter 1907; Dolling and
gies on the glandular hairy plant surfaces are suggested for Palmer 1991; Ellis and Midgley 1996; Anderson and
Midgley 2002; Anderson 2006). The host plant is densely
covered with glandular trichomes of three different types
Handling editor: Heikki Hokkanen. (short, middle-sized, and long tentacle-shaped ones; Fenner
1904; Bruce 1907; Voigt et al. 2009). It is a three-dimen-
D. Voigt (&)  S. Gorb sional, meshed, sticky, flypaper trap that has been observed
Evolutionary Biomaterials Group, Department of Thin-Films
to capture numerous insects of considerable body mass
and Biological Systems, Max-Planck Institute for Metals
Research, Heisenbergstraße 03, 70569 Stuttgart, Germany (Marloth 1903, 1910). Nevertheless, P. roridulae live and
e-mail: voigt@mf.mpg.de walk confidently on the sticky plant surface. The surface of
S. Gorb these mirid bugs is covered with an anti-adhesive, greasy,
e-mail: sgorb@zoologie.uni-kiel.de epicuticular layer that prevents the bugs’ adherence to the
plant secretion (Voigt and Gorb 2008) and enables them to
D. Voigt  S. Gorb
live in a digestive mutualism with R. gorgonias (Ellis and
Department of Functional Morphology and Biomechanics,
Zoological Institute, Christian Albrechts University of Kiel, Midgley 1996). Mirid bugs feed on the insects captured by
Am Botanischen Garten 1–9, 24098 Kiel, Germany plant trichomes and defecate on their leaves (Marloth 1903;

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70 D. Voigt, S. Gorb

Loyd 1934). The nitrogen in the bugs’ faeces is absorbed, Table 1 Comparison of morphometric variables previously referred
for nutrition, through the thin leaf cuticle of the plant, since for Dicyphus errans (Wagner 1970/71) and Pameridea roridulae
(Dolling and Palmer 1991)
the plants themselves produce no enzymes to digest
captured insects (Marloth 1910; Lloyd 1934; Ellis and Variable (mm) Dicyphus errans Pameridea roridulae
Midgley 1996; Anderson and Midgley 2002, 2003). Fur-
Body length 4.7 4.9
thermore, juveniles of Pameridea are primary pollinators of
Body width 1.2 2.5
Roridula’s flowers (Anderson et al. 2003).
Length of anterior leg 5.3 6.2
Recent coevolutionary studies on mutualistic interac-
Width of anterior tibia 0.1 0.3
tions resulted in phylogenetic and geographical associa-
Width of anterior femur 0.3 0.8
tions, suggesting cospeciation, coadaptation, and long-term
coexistence in the Roridula-Pameridea complex (Anderson
et al. 2004; Anderson 2006). In host choice experiments
with Roridula dentata L. (Roridulaceae) and Pameridea whereas geometrical variables of trichomes (length and
marlothi Poppius (Heteroptera, Miridae), bugs preferred diameter) significantly influenced the bug’s attachment
host plants over unrelated host plant species and closely ability. Trichomes have been found to provide suitable
related sister species (Anderson et al. 2004). The presence interlocking sites for attachment and locomotion of this
of an intermediate number of P. marlothi caused positive tiny mirid bug. It clings to single trichomes using one part
growth rates in R. dentata (Anderson and Midgley 2007). of the paired claw.
Only a few data about mutualistic effects on bug’s fit- Preliminary observations on P. roridulae let us suggest a
ness and specific adaptations are currently available. Since different behavior strategy. This insect keeps its body close
relationships between insects and plants are highly diverse to the plant substrate and frequently contacts adhesive tri-
associations, they need broad studies of various factors and chome secretion that covers almost the entire surface of
features at different conditions for a better understanding R. gorgonias and adheres occasionally to bristles on the
and evaluation (Thompson 1988). Besides genetic, fossil, mirid bug0 s body surface (Voigt and Gorb 2008). It runs
taxonomic, geographical, ecological, and biochemical quickly between glandular trichomes, moves sometimes
evidence, insights into the functional morphology and with jump-like movements or short flights. However, it
ethology may indicate associations and coadaptations grooms itself infrequently. Compared to D. errans, mor-
(Futuyma and Slatkin 1983). phological differences are also ascertainable in P. roridu-
Similar to R. gorgonias and P. roridulae, relationships lae; the latter appears burlier. Although the length of the
between mirid bugs and carnivorous plants have been body and legs is similar to that in D. errans, the width
reported from representatives of the plant genera Byblis differs between the species and is distinctly larger in
and Drosera in Australia (e.g. Schuh 1995). These bug P. roridulae (Table 1).
species are representatives of the mirid bug subfamilies Possibly, not only the anti-adhesive, greasy, epicuticular
Orthotylinae and Bryocorinae, which are known to be layer enables these bugs to live on the sticky plant surface,
specialised in living on glandular hairy plants (Reuter but also other morphological and behavioral characteristics
1913; Kullenberg 1946; Falkingham 1995; Dolling and contribute to it0 s ability. The stronger build of P. roridulae
Palmer 1991; Wheeler 2001; Sugiura and Yamazaki 2006; may result in a larger muscle mass, due to which mirid
Voigt et al. 2007). Such mirid bugs have been reported to bugs could generate stronger forces during locomotion on
bear elongated, curved, sharp claws and free, rather large the sticky substrate, where they have possibly to free
pseudopulvilli (Reuter 1913; Wagner 1955). Besides themselves from trichomes’ viscous secretion. As a spe-
specialised claws, they have slim bodies as well as long, cialist species, it is strongly associated with its single host
slender legs (Roberts 1930; Southwood 1986; Voigt et al. plant (Anderson et al. 2004; Anderson 2006). Similar to
2007). In contrast to P. roridulae, other related species related species of Miridae, they have to attach properly to
have been observed avoiding contact with sticky glandular the surface, not only during locomotion and feeding, but
plant secretions by means of morphological adaptations. also during copulation, molting and oviposition of eggs
Additionally, mirid bugs may show a particular behav- into the plant tissue (Wheeler 2001).
iour. For example, Dicyphus errans Wolff (Heteroptera, Thus, the question arises, how do P. roridulae bugs
Miridae, Bryocorinae, Dicyphini) stalks along the plant attach and walk on their host plant? Do their attachment
surface touching only trichome tips and stops frequently to devices and attachment ability differ from those of related
groom itself (Southwood 1986; Voigt et al. 2004). For mirid bugs? Are they specialised for the surface of
D. errans, non-glandular and glandular hairy plant sub- R. gorgonias? Previously, pretarsal structures in the genus
strates have been experimentally demonstrated to be Pameridea have been described to be similar to those of the
important attachment substrates (Voigt et al. 2007), genus Dicyphus, bearing short, curved claws, triangular

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Locomotion in a sticky terrain 71

pseudopulvilli and a straight, bristle-like parempodia visually with a stereomicroscope Olympus SZX 12 with a
(Dolling and Palmer 1991). However, up to now, no DF PLAPO 1xPF objective (Olympus Corp., Tokyo,
illustrations of these structures are available in the Japan), and (3) using digital recordings. Images were taken
literature. using (1) a digital single lens reflex camera Canon EOS
The present study should contribute to the better 20D combined with the Canon macro lens EF 100 mm,
understanding of the complex mutualistic relationship 1:2.8, USM and the Canon macro twin lite MT-24EX
between the protocarnivorous plant and the associated (Canon Inc., Japan), and (2) a Nikon Coolpix E995 digital
mirid bugs, in particular at the interface between the plant camera adapted to the stereomicroscope with a C-Mount
surface and insect attachment system. Using light and cryo- adapter and MDC 2 relay lens MXA 29005 (Nikon Corp.,
scanning electron microscopy (cryo-SEM), we analyzed Tokyo, Japan).
the pretarsal structures of P. roridulae. Its forces, generated To analyze the morphology of the mirid bug0 s attach-
in a traction experiment, were measured on various sub- ment system and the interaction with the plant surface, a
strates: glandular hairy adaxial and abaxial leaf surfaces of cryo-SEM Hitachi S-4800 (Hitachi High-Technologies
the bug’s host plant, smooth adaxial and sparsely hairy Corp., Tokyo, Japan) equipped with a Gatan ALTO 2500
abaxial leaves of Rumex obtusifolius L. (Polygonaceae) and cryo-preparation system (Gatan Inc., Abingdon, UK) was
glass. used. Fresh samples of mirid bug legs and plant leaves
were cut out using a razor blade, mounted on metal holders
by Tissue-TekÒ O.C.T.TM Compound (Sakura Finetek
Materials and methods Europe B. V., Zoeterwoude, Netherlands), frozen in the
preparation chamber at -140°C, sputter-coated with gold–
Insects and plants palladium (6 nm thickness) and examined in a frozen state
in the cryo-SEM at 3 kV and -120°C.
Shrubs of R. gorgonias (seeded, potted, 1–3 year old) with To visualize in detail how mirid bugs’ claws interlock
P. roridulae, were obtained from a private glasshouse with trichomes on the host plant surface, leaf samples of
culture (Klaus Keller, Augsburg, Germany). They were R. gorgonias with attached P. roridulae were prepared,
kept under laboratory conditions during experiments clamped perpendicularly on holders, and examined in the
(23.7 ± 1.7°C, 47.3 ± 10.0% RH, 16 h photoperiod), and cryo-SEM as described above. Additionally, from SEM
fed with wingless, adult Drosophila melongaster Meigen micrographs of 10 randomly selected pretarsi of adult
(Diptera, Drosophilidae; Zoo-Schöniger, Stuttgart, specimens (males and females pooled together), the inner
Germany). length of the claw, diameter of a circle fitting the inner
Leaves from plants of Rumex obtusifolius L. (Polygon- curvature of the claw, and length of the setiform parem-
aceae) at eight-leaf stage were collected from the wayside podia were estimated, using Sigma Scan Pro 5 (SPSS, Inc.,
in a humid mixed forest in Stuttgart-Büsnau (Baden- Chicago, IL, USA) software.
Württemberg, Germany). Its adaxial leaf surface is uneven
because of the prominent venation and irregularly shaped Traction force experiment
convex epidermal cells and has a maximum height of about
9 lm (Holloway 1967; Gorb and Gorb 2009). The cells are Traction force measurements were carried out according to
relatively large (50.0 ± 9.04 lm long, 30.7 ± 7.45 lm Gorb et al. (2004) and Voigt et al. (2007). Insects were
wide), covered with a cuticle and a smooth, amorphous, anaesthetised with CO2 and attached to a 10 cm long human
epicuticular wax layer (0.1 ± 0.05 lm thick). Sporadi- hair with a molten wax droplet. Their forewings were glued
cally, very sparse small wax crystals occur in some places together. Bugs were weighed using an analytical balance AG
on the surface. The crystals have irregular scale-like or 204 Delta Range (Mettler Toledo GmbH, Greifensee,
granular shapes and vary in size (0.5 ± 0.16 lm long, Switzerland). Following anaesthesia, insects were allowed
0.3 ± 0.07 lm wide, and 0.1 ± 0.0 lm thick) (Gorb and to recover for 1 h. Then, the free end of the hair was con-
Gorb 2009). The abaxial leaf side is roughly similar to the nected to the force sensor.
adaxial, however, convex cells are striated and tiny (Hol- Fresh leaves were removed from the middle part of
loway 1967). R. gorgonias plants and attached to a horizontal glass plate
using double-sided tape. Pieces (5 9 5 cm2) were cut out
Observations and structural studies with a razor blade from the intercostal leaf areas of the
non-host plant R. obtusifolius. Using a FORT-10 force
The attachment positions of five male and five female transducer (10 g capacity, Biopac Systems Ltd., Santa
freely walking P. roridulae to the plant surface were Barbara, CA, USA), the traction force was measured in
studied (1) visually without optical instruments, (2) males and females of P. roridulae. Each specimen was

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72 D. Voigt, S. Gorb

tested (1) walking distally on the adaxial and abaxial leaf where claw curvature fits the trichome stem, occurred on
surface of R. gorgonias (contact angle of water could not filiform thread-shaped (Fig. 1o), small (Fig. 1v) and mid-
be determined), (2) walking proximally on the adaxial and dle-sized trichomes (Fig. 1t). On thicker tentacle-shaped
abaxial leaf surface of R. gorgonias, (3) walking distally on ones, the position of claws was similar to that on smooth
the non-host plant R. obtusifolius (contact angle of water surfaces (Fig. 1x). Claws moved apart under load, applied
40–70°, surface energy 35.43 mN*m-1; Gorb and Gorb on the flat surface, allowing pseudopulvilli to get a grip on
2009), and (4) walking on a glass surface (contact angle of the plane surface.
water 55°, surface energy 42.9 mN*m-1). The succession Cryo-SEM studies of the pretarsus clarified its position
of different substrates (1–4) was randomly organised dur- on the plant surface (Fig. 2a) and the structure of attach-
ing the experiment. Before the exchange of substrates, bugs ment devices (Fig. 2b–f). Paired curved claws are
were allowed to groom and to walk over KIMTECH- 23.1 ± 6.77 lm (mean ± SD) long and connected to the
ScienceTM precision wipes (Kimberly-Clark Europe Ltd, unguitractor plate at their bases (Fig. 2c). They have
Surrez, UK). slightly tapered tips (claw teeth, 2.6 ± 0.51 lm long) and
Using AcqKnowledge 3.7.0 software (Biopac Systems widened basal lobe-like structures. The latter ones bear
Ltd, Goleta, CA, USA), force–time curves were recorded to paired, trapezium-shaped pseudopulvilli and between them,
estimate the maximal traction force produced by a single two setiform parempodia (13.8 ± 1.0 lm long). Dorsally,
bug during five consecutive runs on a test substrate. On a stronger (guard) seta is visible (Fig. 2f). The claw surface
each plant surface, five males and five females were tested bears longitudinal striae. The inner side of claws appears
individually. In total, 70 individual tests with 10 different sharp-edged. Adhering to a single short trichome, its stem
specimens were carried out (N = 7 substrates, n = 10 is clamped beween the inner side of one claw part and a
individual tests). Kruskal–Wallis one-way ANOVA on single pulvillus (Fig. 2d, e). The average diameter of an
ranks followed by all pair-wise multiple comparision pro- ideal circle fitting the concavity of claws is 16.1 ±
cedures (Dunn’s test) was applied to determine differences 4.14 lm.
in forces between test surfaces (software SigmaStat 3.1.1Ò,
Systat Software, Inc., Richmond, California, USA). Force Traction force of Pameridea roridulae
measurements in various walking directions on the same
leaf surface of R. gorgonias were included because Plant substrates used in the traction force experiment
trichomes are oriented distally and thus some anisotropical varied in the structure of their surfaces (Fig. 3). The adaxial
effects could occur. In contrast to R. gorgonias, the dock leaf side of R. gorgonias is densely covered with filiform,
plant R. obtusifolius represents a non-host plant having a thread-shaped, distally pointing trichomes and interdis-
totally different surface. Since no structural anisotropy was persed short glandular trichomes (Fig. 3a). The abaxial leaf
found on dock plant leaf surfaces, traction was tested only side bears short and medium-sized glandular trichomes, and
for mirid bugs walking distally on the leaf. the spaces between them are rather smooth, having slight
cell irregularities (Fig. 3b). Leaf margins are densely
covered with long, medium-sized and short trichomes.
Results Leaves of R. obtusifolius appear adaxially predomi-
nantly smooth, slightly textured by convex cells having
The attachment system of Pameridea roridulae sparse cuticular foldings (Fig. 3c), and a few discrete pat-
ches of epicuticular crystal wax granules and stomata
Mirid bugs were able to move quickly and confidently, and (Fig. 3d–e). On the abaxial leaf side, convex cells and
to attach properly to the whole plant surface (stems, both venation are more prominent. Larger veins are covered
leaf sides and inflorescence; Fig. 1a–h). Frequently they with papilla-shaped trichomes (Fig. 3f). Numerous stomata
were observed resting in axillae (Fig. 1b, d, e). Predomi- are found (Fig. 3g).
nant interlocking sites were trichome stems, namely their The body mass of P. roridulae ranged from
bases, up to the lower one-third of the trichome (Fig. 1f–n, 6.7 ± 0.35 mg (mean ± SD) in males to 7.0 ± 0.63 mg in
p, q–s, u, w, y, z). Infrequently, claws clung more distally females without a statistical difference between the sexes
to the trichomes (Fig. 1v). The leg span (distance between (t-test, t = -0.864, P = 0.413, n = 10). Traction forces
left and right leg pretarsi) fits approximately to the width of differed significantly depending on the surface (Fig. 4).
the leaf lamina. Thus, bugs grip trichomes at both leaf The highest forces, up to 8.8 mN, were generated on the
margins at once, and hold their body above the leaf lamina. adaxial leaf surface of R. gorgonias, corresponding to 126
However, bugs also attached to the smooth plant epidermis times the bug’s body mass. The lowest forces were mea-
between trichomes, positioning their tarsus almost parallel sured on the adaxial leaf side of R. obtusifolius and on a
and very close to the surface (Fig. 1f). Claw interlocking, glass surface (47 times the bug’s body weight). Leaf side

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Locomotion in a sticky terrain 73

Fig. 1 Images of various positions of Pameridea roridulae bugs and instar nymph demonstrating similar behavior as in f (h), and pretarsi
their tarsi on the surface of the host plant Roridula gorgonias. a–c A interlocked with trichomes (i, j). k–z Details of pretarsi adhering to
female bug, adhering to trichomes, close to a captured blow fly (a); at the plant surface at various positions. Note: claw interlocking close to
the plant stem in an axilla (b); and close to the leaf tip (c). d, e The the base of a single trichome (k, l, q, p, r, u, x, y, z), at higher sites of
male bug in diverse similar attachment positions. f A female, adhering the trichome stem (t, v), and attachment to the smooth plant surface
to the smooth plant surface between trichomes; pretarsi almost between trichomes (m, n, s). The body length of the bugs is about
orientated parallel to the plant surface. g A female and 5th instar 5 mm
nymph clinging to the plant surface at opposite leaf sides. h–j 5th

and pulling direction did not significantly influence the touches adhesive plant secretion droplets since it adheres,
bug’s performance on R. gorgonias. On R. obtusifolius, predominantly, to the trichome base at the lower third of
higher forces were generated on the rougher abaxial sur- the trichome stem. Thus, the body is held very close to the
face. Exept for the distal direction on the abaxial leaf side plant surface, unlike related species. For example, Dicy-
of R. gorgonias, the adaxial leaf side of R. obtusifolius and phus errans adheres to the glandular surface of Ononis sp.,
the glass surface, no statistical differences were found positioning the long tibiae and the especially long hind
between sexes on the same surface (Table 2). femuri almost vertically to the plant surface (Southwood
1986). Only the tarsal apexes come in contact with the
plant surface and the body is held far away from it.
Discussion

Mirid bug behaviour Pretarsal adaptations to the trichome-covered terrain

The locomotion mode of P. roridulae and the manner in Although the pretarsus of P. roridulae bears similar
which it attaches to plants affirm previous reports that this structures found in related bryocorine bug species (Schuh
bug does not avoid contact with the sticky sites of the host 1976; Cobben 1978; Wheeler 2001), claws in P. roridulae
plant surface (Voigt and Gorb 2008). Rather it frequently are distinctly shorter (mean length 23.1 lm) and thicker,

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74 D. Voigt, S. Gorb

Fig. 2 Cryo-SEM micrographs of the attachment devices in are pressed against the substrate beneath the outspread claws. The
Pameridea roridulae bugs. a Female’s abdominal tip and hind legs, guard seta is clearly visible. (bs) basal lobe-like structure; (cl) claw;
adhering to the glandular hairy abaxial plant surface of Roridula (co) grooming comb; (ct) claw tooth; (fe) femur; (gs) guard seta; (mt)
gorgonias. b Female tarsus. c Female pretarsus (unguitractor plate middle-sized trichome; (ps) pseudopulvillus; (pt) pretarsus; (st) short
hidden by tarsomere 3 and other pretarsal structures). d, e Male trichome; (ta) tarsus; (t1) 1st tarsomere; (t2) 2nd tarsomere; (t3) 3rd
pretarsus interlocked with a short glandular trichome. The middle of tarsomere; (th) trichome glandular head; (ts) trichome stem; (tt)
the trichome stem is clamped between a single claw, its basal lobe- tentacle-shaped trichome; (ti) tibia; (tj) tibia-tarsus joint. Scale bars:
like structure and the pseudopulvillus. f A male pretarsus attached to a = 1,000 lm; b, d = 50 lm; c, e, f = 20 lm
the smooth metallic surface of a SEM sample holder. Pseudopulvilli

e.g., compared to those of the mirid bug D. errans (mean D. errans, where long and thin trichomes with a high
length 74.0 lm, Voigt et al. 2007). The body length is aspect ratio resulted in lower traction forces (Voigt et al.
similar in both mirid bug species (Table 1), but the body of 2007).
P. roridulae appears stronger and thicker. The mean female The heavier body of P. roridulae led us to assume a
body mass in P. roridulae is 7.0 mg corresponding to 1.8 generally larger portion of muscles enabling this bug spe-
times that of the female D. errans. The body mass, in cies to generate stronger forces compared to lighter related
combination with trichome stem flexibility, may explain species. The assumption is confirmed by traction forces
why the bugs P. roridulae predominantly cling to the basal measured with P. roridulae on various surfaces. Traction
part of trichomes. Knowing trichome spring constants and force corresponds to the friction between insect pretarsi
bending behaviour (tentacle-shaped: 0.02 Nm-1, bending and the substrate. The proper adherence to trichomes and
at the tip; medium-sized: 0.08 Nm-1, bending at the tip; applying a pulling force to them while remaining in a
short: 0.95 Nm-1, bending at the base Voigt et al. 2009), certain position, is important in behavioural situations
the trichome’s deflection, caused by the load of a mirid where mirid bugs’ bodies move back and forth or up and
bug, can be estimated. Since the geometry of trichomes is down the substrate (Kullenberg 1946; Voigt et al. 2004;
similar, the deflection of short trichomes may also be Voigt 2005). (1) During sucking the head and thorax are
considered as for the base of tentacle-shaped ones. repeatedly moved up and down, supporting the rostrum
Assuming a resting female (7 mg) having all six legs bending inside the prey. (2) During the final phase of
clinging to trichomes, a tentacle-shaped trichome would copulation the male and female couple their abdominal
bend 1 mm at the tip but only about 20 lm at the base. For tips, heads facing in opposite directions, and move their
comparison, a female D. errans (3.8 mg) would cause a bodies back and forth, as in a ‘‘tug of war’’. (3) The ovi-
trichome deflection of 250 lm at the trichome’s tip and position requires strong up and down movements of the
about 7 lm at the base. Less trichome bending allows more female abdomen and therefore strong attachment to the
effective bug attachment, as shown in previous studies on plant substrate in order to insert the saw-shaped ovipositor

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Locomotion in a sticky terrain 75

Thus, smooth sites on R. gorgonias (trichome-free spaces

on the leaf and wide, non claw-fitting trichome stems)
provide suitable attachment substrates for P. roridulae
bugs. Considering the lowest generated force on a smooth
glass surface (females: 0.7 mN, median), a bug may still
free itself from 10 tentacle-shaped trichomes adhering
coincidentally to the bug’s epicuticle (median adhesion
force of a single trichome: 0.07 mN, Voigt and Gorb
2008).
Pameridea roridulae bugs were observed predominantly
interlocking to trichomes using claws. Although their claws
are much shorter (average length 23 lm) than those of
D. errans (74 lm), the average diameter of an ideal circle
fitting the concavity of claws in both species is about
16 lm. Since mean stem diameters of trichomes on the
surface of R. gorgonias are from 75 lm in median-sized to
120 lm in tentacle-shaped ones (Voigt et al. 2009), claws
of P. roridulae may only interlock with some sites of these
trichome stems. Nevertheless, on rough surfaces, higher
traction forces were measured compared to smooth ones.
For example, on the abaxial leaf side of R. obtusifolius,
small papilla-shaped trichomes provided interlocking sites
for claws and therefore, a stronger traction force of mirid
bugs was recorded than on the smooth adaxial leaf side.
P. roridulae bugs performed best on their host plant. Since
forces did not differ significantly between leaf sides and
pull direction, leaves of R. gorgonias, in general, can be
assumed to be a suitable attachment substrate for male and
female bugs.
Fig. 3 Cryo-SEM micrographs of plant substrates used in traction
force experiment with Pameridea roridulae bugs. a, b The leaf
surface of Roridula gorgonias bears various types of trichomes on the
Sexual dimorphism in traction force generation
adaxial (a), and on the abaxial leaf side (b). c–e The uneven adaxial
leaf surface of Rumex obtusifolius shows irregular convex cells and Compared to females, males generated significantly higher
stomata (c, e), sparse cuticular foldings (arrow) and wax granules (d). forces on the abaxial leaf side of R. gorgonias, on the
f, g The abaxial leaf surface is similar to the adaxial, however,
prominent veins are covered with small papilla-shaped trichomes (b).
adaxial leaf side of R. obtusifolius, and on glass. Differ-
(ft) filiform, thread-shaped trichomes; (mt) middle-sized trichomes; ences between sexes have been previously found in traction
(st) short trichomes; (tt), tentacle-shaped trichomes. Scale bars: a, b, force experiments with leaf beetles Chrysolina polita L.
c, f, g = 50 lm; d = 1 lm; e = 20 lm (Coleoptera, Chrysomelidae; Stork 1980) and ladybird
beetles Coccinella septempunctata L. (Coleoptera, Cocci-
nellidae; Gorb et al. 2008). Males of both species adhere
into the plant tissue. (4) Molting nymphs and emerging stronger to smooth surfaces, whereas females performed
imagines free themselves from old cuticle by pulling their better on rough ones. However, attachment systems of
body distally. For P. roridulae, the challenge is to perform beetles (hairy) and mirid bugs (smooth) differ completely
these actions even upon its adhesive host plant terrain. and cannot be compared outright in the context of our
Possibly, for this reason P. roridulae may generate higher results (Beutel and Gorb 2001). Since significances
traction forces (126 times higher than the body mass) than between male and female force values occur on predomi-
the related D. errans (34.2 times higher than the body nantly smooth substrates, one may conjecture that differ-
mass) on both a glandular hairy plant surface as well as on ences in material properties between male’s and female’s
glass (P. roridulae: 47, and D. errans: 6.1 times higher pseudopulvilli exist. These pad-like attachment structures
than the body mass). The latter values indicate that mirid in mirid bugs, adhering to smooth surfaces, need further
bugs may also adhere sufficiently to microstructured and micromechanical analyses for better understanding of their
smooth surfaces such as those of R. obtusifolius and glass. functionality depending on sex and species.

123
76 D. Voigt, S. Gorb

A B
10
distal

R. gorgonias
adaxial
proximal
8
Traction force [mN]

distal
abaxial
proximal
6

R. obtusifolius
adaxial distal

abaxial distal
4
glass

proximal

proximal
distal

distal

distal

distal

glass
2

abaxial
adaxial

adaxial

abaxial
0
distal proximal distal proximal distal distal
adaxial abaxial adaxial abaxial
Roridula gorgonias Rumex obtusifolius glass R. gorgonias R. ob tusifoli us
Surface

Fig. 4 Forces generated by Pameridea roridulae in traction experi- Asterisks show statistical differences between males and females (for
ments on leaf surfaces of Roridula gorgonias (traction in distal and statistical differences see Table 2). b Statistical differences between
proximal direction) and Rumex obtusifolius (traction in distal direc- surfaces (Kruskal–Wallis one-way ANOVA on ranks, for males:
tion), and on cleaned glass (control). a Box-and-whisker diagram H6,173 = 101.5, P B 0.001; for females: H6,173 = 111.7, P B 0.001;
showing the traction force of single mirid bugs measured on various all pairwise multiple comparison procedures, Tukey test, P \ 0.05 for
surfaces. The ends of the boxes define the 25 and 75th percentiles, with both, separately males and females)
a line at the median and error bars defining the 10 and 19th percentiles.

Table 2 Statistical differences of forces between males and females, the substrate due to the absence of detailed analyses.
generated in the traction experiment on various surfaces (N## = 5, Considering published data on both D. errans (Southwood
N$$ = 5, n = 5). See also Fig. 2
1986; Voigt et al. 2007) and P. roridulae (Voigt and Gorb
Surface and traction direction Statistical values 2008) in combination with results obtained in the present
Roridula gorgonias, t = 1.020, P = 0.313 study, we may hypothesize two different strategies of
adaxial leaf side, distal locomotion and attachment on glandular hairy plant sur-
Roridula gorgonias, t = 1.797, P = 0.079 faces in species within one and the same mirid tribe
adaxial leaf side, proximal Dicyphini of the subfamily Bryocorinae (Fig. 5). (1) The
Roridula gorgonias, T = 761.000, P = 0.017 ‘‘avoidance strategy’’, as used by D. errans, consists of
abaxial leaf side, distal keeping the body at a large distance from the plant surface
Roridula gorgonias, t = 0.477, P = 0.636 and glandular secretions of trichomes (Fig. 5a). This
abaxial leaf side, proximal
strategy relies on a light and slim body, long and slender,
Rumex obtusifolius, t = 2.893, P = 0.006
carefully-moving legs, whose tibiae are oriented almost
adaxial leaf side, distal
vertically to the substrate. Pretarsi are equipped with long,
Rumex obtusifolius, t = 0.561, P = 0.578
abaxial leaf side, proximal narrow claws. Moreover, frequent resting and grooming are
Glass T = 756.000, P = 0.022 observed in these bugs, indicating that they do not have a
protective layer on their epicuticle maintaining the
t computed t-statistic in the t-test for normally distributed data, T
body clean and free from adhesive plant secretion. (2)
computed T-statistic in the Mann–Whitney rank sum test for not
normally distributed data, P computed p-values for t and T, P. roridulae has a ‘‘defense strategy’’ allowing close con-
respectively tact with the plant surface and the trichomes’ adhesive
secretion (Fig. 5b). The body is heavy, with relatively thick
Two locomotion strategies of mirid bugs legs that are predominantly used for attachment to the
trichome bases by short claws. Mirid bugs with ‘‘defense
Generally, in species of the order Heteroptera, a notable strategy’’ generate stronger forces during locomotion,
diversity of pretarsal structures has been reported and enabling them to walk, unhinderd, on a plant surface
discussed in the evolutionary context (Schuh 1976; Cobben covered with sticky trichomes and to free themselves from
1978). Some peculiar host plant relations have been pre- adhesive secretions. Additionally, the greasy secretion,
viously assumed, but the adaptive value of these structures found in the P. roridulae epicuticle, contributes to reducing
is difficult to interpret in direct functional connection with the adhesion of the plant adhesive to the bug. One may

123
Locomotion in a sticky terrain 77

conjecture that the defense strategy is a specific adaptation Rhopalidae) appear stronger and walk unhamperedly
of bugs P. roridulae for a life on the glandular plant surface between trichomes like a ‘‘snowplow’’ on the same plant
of R. gorgonias. This would support recently published surface (Schwoerbel 1956). Thus, for non-mirid species,
coevolutionary considerations on cospeciation, coadapta- the avoidance strategy is not only characteristic of gener-
tion, and long-term coexistence in the Roridula-Pameridea alists, because the specialist G. punctipes is obligatory
complex (Anderson et al. 2004; Anderson 2006). On the associated with O. spinosa.
other hand, the avoidance strategy could be a typical fea-
ture of generalists, like D. errans, having a broad range of
host plants. Concluding remarks
To what extent the hypothesized strategies may be the
case for other mirid bug species, is difficult to appraise, Pameridea roridulae lives in a complex, three-dimensional,
because neither detailed comparative nor experimental sticky terrain where it moves without hindrance. In contrast
studies have been previously carried out. Representatives to related species within the tribe Dicyphini of the family
of the mirid bug subfamilies Orthotylinae and Bryocorinae Bryocorinae, P. roridulae do not avoid adhesive secretions
are generally known to be specialised in living on glandular but rather walk between glandular trichomes and touch
hairy plants (Reuter 1913; Kullenberg 1946; Cassis 1984; them very frequently. Previously, an anti-adhesive epicu-
Falkingham 1995; Dolling and Palmer 1991; Wheeler ticular layer protecting mirid bugs from being captured by
2001; Sugiura and Yamazaki 2006; Voigt et al. 2007). the adhesive plant secretion has been described. The
Several of them even live on insectivorous plants of the present results show that the mode of attachment and
genera Drosera and Byblis (e.g. China and Carvalho 1951; locomotion on the plant surface also contributes to the
China 1953; Southwood 1986; Falkingham 1995). These specialisation of P. roridulae to a life on the protocar-
bugs also feed on trapped insects, but unlike P. roridulae, nivorous flypaper trap R. gorgonias. There are direct,
they avoid the contact with the sticky surface of their host biomechanical interactions at the interface between the
plants (Russell 1953). plant surface and insect attachment system. Dicyphine bugs
Only one previous report on different modes of locomo- use claws to cling to trichomes and use pad-like pseudo-
tion on glandular plant surfaces is found in literature, how- pulvilli for adhesion to smooth hairless patches or to thick
ever, for other true bug families than Miridae (Schwoerbel trichome stems. This strategy allows them to generate
1956). Stilt bugs Gampsocoris punctipes Germ. (Heteroptera, distinctly higher traction forces if compared to other mirid
Berytidae) avoid the contact with glandular plant secretion bug species. Thus, P. roridulae may even overcome con-
of the spiny restharrow Ononis spinosa L. (Fabaceae) by tact with the adhesive secretion of numerous trichomes.
means of their slim, small body and long, slender legs, Based on available data, we have hypothesized, here, two
whereas juveniles Corizus hyoscami L. (Heteroptera, strategies of locomotion and attachment on glandular hairy

A B

plant surface plant surface

avoidance defence

Fig. 5 Two assumed strategies of mirid bugs of the tribus Dicyphini their body clean. b ‘‘Defense strategy’’: the heavy, tough body is
(Heteroptera, Miridae, Bryocorinae) walking on plant surfaces, which situated close to the plant surface, frequently contacting trichomes
are densely covered with glandular trichomes. a ‘‘Avoidance and their viscous secretion. Bugs overcome the adhesion of the plant
strategy’’: the light, slim body is held at a large distance from the surface by generating strong forces during locomotion and having a
plant surface and adhesive secretion of trichomes by using long, thick anti-adhesive epicuticular greasy layer on their cuticle
slender legs. Additionally, bugs frequently rest and groom to keep

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78 D. Voigt, S. Gorb

plant surfaces within the mirid bug tribus Dicyphini: (1) from a revised ordinal phylogeny. J Zool Syst Evol Research
avoidance strategy, characterised by the slim body held at a 39:177–207
Bruce AN (1907) On the distribution, structure, and function of the
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Cobben RH (1978) Evolutionary trends in Heteroptera. Part II.
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acknowledged for provision of plants and bugs. The bug species was Futuyma DJ, Slatkin M (eds) Coevolution. Sinauer Associates,
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Kastner (Max-Planck Institute for Metals Research, Stuttgart, Ger- Gorb E, Gorb S (2009) Effects of surface topography and
many) provided linguistic corrections of the manuscript. This study chemistry of Rumex obtusifolius leaves on the attachment of
was supported by the Federal Ministry of Education and Research, the beetle Gastrophysa viridula. Entomol Exp Appl 130:
Germany (BMBF project Inspirat 01RI0633D). 220–222
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Creative Commons Attribution Noncommercial License which per- digestive zone of the pitcher in the carnivorous plant Nepenthes
mits any noncommercial use, distribution, and reproduction in any ventrata and its role in insect trapping and retention. J Exp Biol
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