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Ent. exp. & appl. 28(1980) 100~107. Ned. Entomol. Ver.






Department of Zoology, University of Manchester, Oxford Road, Manchester, MI3 9PL, U.K.
Mustard beetles (Phaedon cochleariae (Fabricius)) adhered slightly better to hairy leaves than to
glaucous leaves of Brassica oleracea but by far the best adhesion was to glossy leaves. Scanning electron
mierographs illustrate how adhesion between the climbing setae on the tarsi of P. cochleariae and the
leaf surfaces was hindered by the vertical rods and dendritic plates on glaucous leaves. The importance
of the wax bloom in determining colonisation of brassicas by insects is discussed.

Many research workers in the field of plant protection have concentrated their
efforts on the physiological adaptations of plants against insect predation. Much
less research has centred on how morphological features of the plant cuticle
protect against insects. Recently, Levin (1973) has reviewed the literature on the
r61e of trichomes in plant defence. Observations on the tarsi of a large number of
plant beetles and the results of experiments on the mode of adhesion (Stork 1977,
1980a, b) show that setae on the tarsi are adapted for adhesion to smooth surfaces.
Insect resistance associated with high densities of leaf trichomes, such as is found
in wheat to the cereal leaf beetle Oulema melanopa (L.) (Chrysomelidae;
Coleoptera) (Gallun et al. (1966), Schillinger & Gallun (1968) and Ringlund &
Everson (1968)) may at least in part be based on the trichomes preventing the tarsi
(and their setae) contacting the underlying leaf surface, so hindering adhesion.
Baker (1972, 1974) using an S.E.M. showed that the wax bloom on a glaucous
Brussels sprout leaf consists of long vertical tubes with an interweaving overlay of
dendritic plates, whereas that on a glossy mutant (G13) leaf consists of a flat
amorphous layer with some small mounds. Many functions of the wax bloom have
been recognised, including the prevention of water loss, water repellency,
reduction of surface damage and abrasion and reduction of chemical and
pathogenic attack. This paper investigates whether the form of the wax bloom on
Brussels sprouts (Brassica oleracea var. gemmifera) may also serve to reduce the
adhesion obtained by Phaedon cochleariae (Chrysomelidae; Coleoptera), the
mustard beetle.
~) Present address: Department of Entomology, British Museum (Natural History), Cromwell Road,
London, SW7 5BD, U.K.




Inclined plane. Plants of three cultivars of Brassica oleracea were grown in a

greenhouse from seed supplied by the Scottish Horticultural Research Institute.
The phenotypes of these cultivars were as follows: No. A4124, glossy mutant G 13,
B. oleracea vat. gemmifera, No. A4175, glaucous (normal), B. oleracea vat.
gemmifera, No. 727/76 HRD, hairy, B. oleracea (of complex parentage; the glossy
mutant differing from the glaucous (normal) on a single gene).
A leaf of about 120 cm: surface area from a 10-week-old plant of one of the
three brassica cultivars was attached by its abaxial surface to a perspex plate
whose bottom edge was hinged such that it could be inclined at any angle from
~ A single adult beetle was placed on the leaf whilst the plate was
horizontal. Once the beetle started walking, the angle of inclination of the perspex
plate was slowly increased from 0 ~ to 180~ If the beetle fell off, the angle of
inclination was recorded. The procedure was repeated on a leaf of each of the
other two cultivars and for fourteen other adult beetles. The whole experiment
was then repeated a further five times using the same beetles but with fresh leaves
for each series of trials. Although not normally found on Brussels sprouts,
P. cochleariae is a common pest of some other brassicas such as turnips. It is also of
a suitable size for use in the S.E.M., and is easily obtained. The specimens used in
these experiments were cultured in the laboratory at about 27 ~, on turnip plants.
S.E.M. of live beetles on fresh plants. Fresh leaf discs were stuck (abaxial surface
downwards) with double-sided cellotape to S.E.M. stubs without touching their
adaxial surfaces. An adult live P. cochleariae was attached by its elytra to a thin
metal rod about 15 mm long using a beeswax-resin mixture (7:4) (Stork, 1977). The
bent ends of the rod were then glued to the sides of the S.E.M. stub so that the
beetle was in 'normal' contact with an undamaged region of the leaf (Fig. 1). The
stub was placed in a Cambridge S150 S.E.M. and the contact between the living
beetle's tarsi and the fresh plant surface observed. Tarsal contact with leaves from
both the glossy and glaucous cultivars were compared.

Inclined plane. The angle of inclination at which the beetles fell off the three leaf
surfaces is recorded in Fig. 2 where the data are grouped in 10~ intervals (e.g.
~ and 31~176
The angles at which beetles fell off the glaucous leaves
varied between 68 ~ and 20 ~ but was generally between 30 ~ and 50 ~. In nearly all
the trials with glossy leaves the beetles adhered through to 180~. The beetles
generally fell off hairy leaves at angles similar to those for glaucous leaves but in a
few trials they were able to adhere above 60 ~ and some remained attached at 180~.
Trichomes were found mainly in the distal half of the adaxial surface of the hairy
leaves at a maximum of about 25 trichomes/cm:. Individual beetles did not differ
significantly in their ability to adhere. Only the data for the glaucous leaves were

Fig. 1. (A) Micrograph of live Phaedon cochleariae attached to metal bar (part in view, indicated by
arrow) by wax/resin mixture. Bar attached to sides of S.E.M. stub~ Several tarsi adhering to surface of
leaf-disc on stub. Scale bar = 1 ram, (B) Region in (a) showing contact of adhesive setae (arrow) on the
tarsi with leaf surface. Scale bar = 250 ~m.

R O L E OF W A X B L O O M 5 IN A T T A C H M E N T T O B R A S S I C A S BY P H A E D O N C O C H L E A R t A E











8b 100 ;~0 T~o 1~0 1~0

Angle of inclination
(b) 80




z 30














Angle of inclination











100 120

Angle of inclination
Fig, 2. Histograms of inclination angles at which beetles fell off three leaf surfaces.




normally distributed (Fig. 2) and therefore the data for the three leaf surfaces were
compared by the non-parametric Kolmogorov-Smirnov two-sample (two-tailed)
test (Siegel, 1956), Results for the glaucous glossy and hairy leaves all differed
significantly (Table I).

S.E.M. of live beetles on fresh plants. Observation of live beetles on fresh plant
material in the S.E.M. was found to be considerably more difficult than that of
prepared dried specimens. In addition to delays in pumping out the specimen

Comparison of inclination angles ( D ) at which P. cochleariac fell off three leaf surfaces using the
Kolmogorov-Smirnov two- sample (two- tailed) test

Glaucus v Hairy
Glaucus v Glossy
Glossy v Hairy

Level of signif.
difference (%)



chamber caused by the large amount of moisture present, using uncoated

specimens resulted in problems with charging, and beam damage such as melting
cuticular wax and adhesive s'etae. However, although the beetles seldom moved at
all in the S.E.M., most recovered afterwards. Figs 3A--C show the tips of the
adhesive setae in close contact with a glossy leaf. The leaf surface is fairly smooth
and the adhering regions of the seta extensive. In contrast, the contact between
the setae and glaucous leaves is poor (Figs 3D--F), the former being covered with
clumps of powdery debris and the latter bearing a dense wax bloom apparently
consisting of almost vertical rods and dendritic plates at various angles.

As the gross morphology of the surfaces of the glossy and glaucous leaves was
similar, the differences observed (Fig. 2) in the ability of the beetles to adhere to
them almost certainly arose from differences in the finer structure of their cuticles.
Stork (1977, 1980b) proposed that adhesive setae have evolved to adhere to
smooth surfaces. The tubes and dendritic plates of wax on the glaucous leaves
present a much rougher and coarser topography than the relatively smooth
amorphous layer of wax on the glossy leaves and, as expected, adhesion was
greater on the latter than on the former. In Figs 3A--C large areas of the tips of
the adhesive setae of P. cochleariae are in contact with the surface of the glossy leaf
whereas in Figs 3 D - - F there is almost no contact between the setae and the
glaucous leaf surface. Furthermore, the adhesive setae in the latter micrographs
are covered with clumps of debris. This is probably epicuticular wax removed as
the setae are drawn ~tcross the leaf. The removal of some of the wax in this
manner, has left some bare patches on the leaf surface, as can be seen in the
bottom right-hand corner of Fig. 3D. Thus, although it is possible that differences

Fig. 3. Adhesive setae in contact with surface of glossy (A--C) and glaucous (D--F) leaves. Wax t~loom
on glossy leaves relatively smooth with a few mounds; on glaucous leaves comprised of tubes and
dendritic plates. Note clumps of powdery material on setae in D--F. Scale bars = (A) 10~tm, (B) 4~tm,
(C) 10~tm, (D) 10gm, (E) 10~tm, (F) 20tim.



in the chemical composition of the epicuticular waxes on glaucous and glossy

leaves may have affected the adhesion attained by the beetles in the experiment
above, the differences in their physical condition are more likely to have achieved
this result.
Glossy-leaved cultivars of B. oleracea have been found to be preferred over
glaucous-leaved forms by several flea beetle species, Erioischia brassicae (Diptera),
Myzus persicae (Aphidae), coccinellids and anthocorids (Way & Murdie (1965) and
Anstey & Moore (1954)). All these results may be caused by these insects adhering
better to glossy than~to glaucous leaves as all these insects (except M. persicae) use
adhesive setae to hold on to plants. In contrast, glaucous B. oleracea was more
susceptible to the cabbage aphid Brevicoryne brassicae and to the cabbage whitefly
Aleyrodes brassicae, than glossy cultivars (Way & Murdie, 1965; Thompson, 1963).
Both these insects grip with fleshy pads (similar to those on the tarsi of
cockroaches, as described by Arnold (1974) instead of adhesive setae. Perhaps the
individual preference of each of these insects for glaucous and glossy brassicas is
determined, at least in part, by its mode of adhesion.
The wax blooms on the glaucous and hairy leaves were virtually identical (i.e.
glaucous) and in many of the trials the beetles appeared to have as much difficulty
adhering to the hairy as to the glaucous plants. This is reflected in the similarity of
the left-handside of their histograms (Fig. 2). However, the histograms do differ
significantly and in some trials the beetles were able to adhere beyond 60 ~ on the
hairy plants. In these trials the trichomes, rather than complementing the affect of
the glaucous wax bloom in reducing the adhesion of the adhesive setae, probably
provided anchorage sites for the tarsal claws.
On the basis of the above conclusions, it is not surprising that many commercial
brassicas have non-pubescent and glaucous foliage.
I thank Professor E. R. Trueman for facilities and Dr. A. B. Wills, Scottish
Horticultural Research Institute, for the brassica seeds; Dr. E. A. Baker (Long
Ashton Research Station, Bristol University) for advice on plant waxes, and Dr.
M. E. G. Evans for discussion. Carried out under a Science Research Council
postdoctoral award.
Senfk~.fer (Phaedon cochleariae) konnten sich etwas besser an.haarigen Blfittern von Brassica oleracea
halten als an wachsigen Blfittern, weitaus am besten war abet die Adhfision an glatten Blfi.ttern
(P<0.05). Scanning-elektronenmikroskopische Aufnahmen zeigen, wie die Adh/ision zwischen den
Steigborsten der Tarsen yon P. cochleariae und den Blattoberflfichen behindert wurde durch die
vertikalen Leisten und durch die dendritischen Platten auf den wachsigen BlS.ttern. Die Bedeutung der
Wachsausscheidungen ffir die Kolonisation von Brassica durch Insekten wird diskutiert.

ANSTEY, T. H. & MOORE, J. F., 1954. Inheritance of glossy foliage and cream petals in Green Sprouting
Broccoli. J. Heredity, 45 : 39--41.



ARNOLD, J. W., 1974. Adaptive features on the tarsi of cockroaches (lnsecta: Dictyoptera). Int. J. Insect
Morphot. Embryol., 3 : 317--334.
BAKER, E. A., 1972. The effect of environmental factors on the development of the leaf wax of Brassica
oleracea vat. gemmifera. MSc. thesis, University of Bristol.
-1974. The influence of environment on leaf wax development in Brassica oleracea vat. gemmifera.
New Phytologist, 73:955--966.
GAk~LUN,R. L., RUPPEL, R. & EVERSON, E. H., 1966. Resistance of small grains to the cereal leaf beetle.
J. econ. Entomol. 59 : 827--829.
LEVIN, D. A., 1973. The role of trichomes in plant defense. The Quarterly Review Biol., 48 : 3--15,
RINGLUND, K, & EVERSON, E. H., 1968. Leaf pubescence in common wheat, Triticum aestivum L., and
resistance to the common leaf beetle, Oulema melanopus L. Crop Science 8 : 705--710.
SCHILLINGER, J. A, & GALLUN, R. L. 1968. Leaf pubescence of wheat as a deterrent to the cereal leaf
beetle, Oulema melanopus. Ann. entomol. Soc. of Amer., 61 : 900--903.
SIEGEL, S., 1956. Nonparametric Statistics for the Behavioural Sciences. London, McGraw-Hill.
STORK, N. E., 1977. The Beetle~Substratum Interface. PhD. Thesis, University of Manchester.
1980a. A Scanning Electron Microscope study of tarsal adhesive setae in the Coleoptera. Zool. J.
Linn. Soc. 68 : 173--306.
-1980b. Experimental analysis of adhesion of Chrysolina polita (Chrysomelidae; Coleoptera) on a
variety of surfaces. J. exp. Biol. (in press).
THOMPSON, K. F., 1963. Resistance to the cabbage Aphid (Brevicoryne brassicae) in Brassica plants.
Nature, Lond. 198 : 209.
WAY, M. J. & MURDIE, G., 1965. An example of varietal variations in resistance of Brussels sprouts.
Ann. appl. Biol., 56 : 326--328.

Accepted: March 8, 1980

Ent. exp. & appl. 28 (1980) 107--108. Ned. Entomol. Ver. Amsterdam

A. K. MINKS & P. GRUYS, (Eds): Integrated control of insect pests in the Netherlands, Centre for
Agricultural Publishing and Documentation, Wageningen, 1980, 304 pp., D.FI. 60.
Stemming from the far-sighted initiative of De Wilde, De Fluiter and others, the Dutch Working
Party on Integrated Control was formed as long ago as 1958. This book marks a "coming of age" for the
work of the group and presents 61 papers by workers in the Netherlands concerned with the
implementation, strategic research and theory of integrated control. The papers are relatively brief but
most bring together and up-date the available information and provide key references for those
interested in further detail. Comprehensive reviews of special fields are not included, but the book does
provide a cohesive story of recent Dutch work which clearly was planned and was aimed at practical
The first three chapters are concerned with the crops that have received most attention: fruit crops
(11 papers), field crops (10) and glasshouse crops (7). Six further chapters are concerned with specific
components that can form part of an integrated approach to insect pest control: the study of insect/host
plant relationships (5 papers) and of host plant resistance to pests (8), pheromones and attractants (3),
selective insecticides (5), insect hormones (4), and micro-organisms, viruses and nematodes (5). There
is a final chapter on supporting techniques which include systems analysis and simulation models (2), a
study of microclimate in an apple orchard (1) and the use of radionuclides and ionizing radiation in
research (1).