Vol. 18, No. 5/6, pp.

289-310, 1989
Int. J. lnsectMorphol. & EmbryoL, 0020-7322/89
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I N T E R N A L A N A T O M Y A N D M O R P H O L O G YO F
FRANKLINIELLA OCCIDENTALIS ( P E R G A N D E )
( T H Y S A N O P T E R A• T H R I P I D A E W
) ITHSPECIAL
R E F E R E N C ETO I N T E R A C T I O N S B E T W E E N T H R I P SA N D
T O M A T O S P O T T E DWILT V I R U S

DIANE E. ULLMAN, DAPHNE M. WESTCOT, WAYNE B. HUNTER

and
RONALDF. L. MAU

Department of Entomology, University of Hawaii, 3050 Maile Way, Rm 310 Honolulu, HI 96822, U.S.A.

(Accepted 20 September 1989)

Abstract--The internal morphology of the western flower thrips, Frankliniella

occidental& (Pergande) (Thysanoptera : Thripidae) (WFF), a primary vector of tomato
spotted wilt virus (TSWV), was shown to bear certain similarities to other thrips species,
particularly in the composition of the piercing-sucking feeding structures. Striking
differences were observed in number, position and ducting of the salivary glands, the
morphology of the alimentary canal and the number and arrangement of the malpighian
tubules between the WFT and other studied Thysanopterans. These differences provide
support for the conclusion that internal morphologies may vary widely in this order, and
perhaps among species in the same genus. The results of our investigation support the
need for more detailed studies of other thrips species, particularly in light of the potential
importance morphological characteristics may play in governing the capacity of thrips
species to serve as vectors of TSWV.

Index descriptors(in addition to those in title): Feeding structures, ultrastructure,

alimentary canal, reproductive organs, electron microscopy.

INTRODUCTION
THRIPS (Order Thysanoptera)
: are diminutive insects that are widespread throughout
the world in habitats ranging from forests, grasslands and scrub to cultivated crops and
gardens (Lewis, 1973). Many of the 5,000 known species (zur Strassen, 1960) are
recognized as economically important insects causing direct damage to plants as a result
of their feeding and in some cases, indirect damage as vectors of plant pathogens (Curtis,
1860; Osborn, 1888; Froggart, 1906; Pittman, 1927; Hansen, 1929; Bald and Samuel,
1931). Perhaps because of their extremely small size, there is a paucity of information
regarding the internal morphology of this large and varied group of insects (Lewis, 1973).
Several studies have focused on the unique asymmetrical morphology of the proboscis or

Correspondence to: Dr. Diane E. Ullman, Department of Entomology, University of Hawaii, 3050 Maile
Way, Room 310, Honolulu, HI 96822, U.S.A. Tel: 808-948-6746.

289
290 et al.
DIANE E. ULLMAN

m o u t h c o n e , the feeding structures housed therein and the head (Reyne, 1927; Risler,
1957; Davies, 1958; Mickoleit, 1963; M o u n d , 1971; Heming, 1978; Chisholm and Lewis,
1984; H u n t e r and U l l m a n , 1989). Only Pesson (1951) and Peterson (1915) have
thoroughly addressed the internal a n a t o m y and morphology of the Thysanoptera. Their
studies of 17 thrips species in the suborders Tubilifera and T e r e br a n t ia demonstrate that
internal morphology varies widely, even amon g species within a given genus.
T h e primary goals of this paper are to describe the internal morphology of
Frankliniella occidentalis (Pergande), the western flower thrips (WFT) on an organismal
and cellular level and to compare it with other previously described species. The W F T is
a direct pest of m a n y crops and perhaps most importantly, a primary vector of tomato
spotted wilt virus (TSWV) (Sakimura, 1960; Paliwal, 1976; A m i n et al., 1981; G r e e n o u g h
et al., 1985; A l l e n and B r o a d b e n t 1986;
, Cho et al., 1987). O ur examination of the
internal morphology of W F T is a first step toward understanding thrips/TSWV
interactions at a cellular level. By comparing the internal morphology of W F T with other
vector and non-vector thrips species we hope to increase our knowledge of potential
mechanisms governing vector specificity and competency.

MATERIALS AND METHODS

We examined the internal morphology of the WFT, Frankliniella occidentalis by light, scanning and
transmission electron microscopy. All thrips examined were adults provided from the colony of Dr. R. F, L.
Man, Department of Entomology, University of Hawaii at Manoa. A Wild M8 dissecting microscope (Wild
Leitz, Wild Leitz USA, Inc., 1123 Grandview Drive, So. San Francisco, CA 94080) was used to view gross
dissections of thrips performed in physiological saline (0.85%). Phase contrast on a Leitz Diaplan compound
microscope (Wild Leitz, Wild Leitz USA, Inc., 1123 Grandview Drive, So. San Francisco, CA 94080) was used
to further view slide squashes of many of these gross dissections. More than 100 male and female thrips were
dissected and examined in this way.
Thick sections of thrips were prepared for viewing with the compound microscope as follows: fixation in 4%
glutaraldehyde (in 0.05 M cacodylate buffer, pH 7.34) for 4-8 hr, 4 washes in 0.05 M cacodylate buffer, post-
fixation for 2-4 hr in osmium tetroxide (in 0.05 M cacodylate buffer), another 4 washes in 0.05 M cacodylate
buffer, dehydration in a gradient series of ethanol over a period of 4-6 hr and finally a propylene oxide
transition and embedding in Spurrs (Spurr, 1969) (Ted Pella Inc, Redding, CA 96099). After embedding, the
thrips were serially sectioned at 1 ~m on a Reichert OMU-2 ultramicrotome in dorsoventral and sagittal
orientations. Sectioned were mounted on glass slides in a drop of water and heated until sections dried down. In
some cases, sections were deresined in 1.5-3% NaOH (in absolute ethanol). This process was repeated at least
3 times. Then slides were placed in phosphate buffer (pH 4) for 5 rain. Slides were further rinsed in double
distilled water (DDW) and allowed to air dry prior to staining. A number of stains, including hematoxylin and
eosin, Giemsa, Mallory's Triple and methylene blue were tried on deresined sections, as well as sections in
resin (Humason, 1967). In both cases, we obtained the most consistent results with methylene blue (modified
from Humphrey and Pittman, 1974). Fourteen individuals, both male and female were thick sectioned and
examined as described.
Thrips were prepared for Scanning Electron Microscopy (SEM) according to the protocol of Hunter and
Ullman (1989). After critical point drying, thrips were placed on SEM stubs with silver conductive paint and
then coated with silver conductive paint. After thorough drying, specimens were cut longitudinally by lightly
dragging a Teflon razor blade across either the dorsal or ventral surface of the insect. The cuticle was then
carefully pealed back with fine forceps to expose internal structures. These specimens were sputter coated
(Hummer II, Technics, 5510 Vine Street, Alexandria, Virginia 22310) and examined with a Cambridge
Stereoscan 150, SEM (Cambridge Instruments, UK) at 20 kV. Thirty-two specimens were dissected, viewed
and photographed using this technique.
Thrips were prepared for Transmission Electron Microscopy (TEM) by cutting live thrips in half with a Teflon
razor blade either between the head and thorax or between the thorax and abdomen in a drop of cold
physiological saline (0.85%) on a glass microscope slide. A few drops of cold 4% ghitaraldehyde (in 0.05 M
cacodylate buffer, pH 7.34) were then added to the dissection. Immediately thereafter, 2-3 drops of 2% agar
(Bacto-agar in 0.05 M cacodylate buffer, 40°C) was layered on top of the dissection. After the agar set, a small
section containing the dissected specimen was excised and fixed for 2-4 hr in a vial of 4% glutaraldehyde (in a
0.05 M cacodylate buffer, pH 7.34). Tissues were processed as described earlier for light microscopy. The
Internal Anatomy and Morphology of Frankliniella occiclentalis 291

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292 DIANE E. ULLMAN
et al.

embedded specimens were then serially sectioned on a Porter Blum MT-2 microtome, 60-80 nm in thickness
and picked up on formvar coated slot grids. Grids were contrasted with 2% uranyl acetate in 50% ethanol for
30-40 rain and post-stained with lead citrate in NaOH for 10-15 rain. Sections were examined and
photographed using a Zeiss 10A transmission electron microscope (Carl Zeiss, West Germany).

RESULTS
The usual opisthorhynchous condition of the Thysanoptera is present in the WFT, with
the clypeus fully ventral and extending posteriorly to the mouthcone, which emerges
perpendicularly to the clypeus (Fig. la). Less than 20 txm within the mouthcone the
paired, interlocked maxillary stylets diverge, marking their juncture with the
precibarium just prior to the cibarium (Fig. la). The precibarial canal channels ingested
plant cell contents into the cibarium, which is positioned ventral to the brain (Fig. la).
As in other piercing-sucking insects, the cibarial pump expands as the cibarial dilator
muscles contract creating the tension required to draw fluids up through the food canal
formed by the stylets and the aforementioned precibarial canal. As the cibarial dilator
muscles relax, the cibarial pump closes and the contents of the cibarium enter the
oesophagus, which travels through the circumoesophageal passage formed by the juncture
between the brain and the suboesophageal ganglion (Figs la, lb, 2). The corpora
cardiaca lies next to the oesophagus within the circumoesophageal passage (Fig. 2).
After emerging from the circumoesophageal passage the oesophagus lies on top of the
suboesophageal ganglion and ventral ganglia up to its juncture with the midgut within the

FIG. 2. SEM micrograph showing dorsal view of brain (Br) and suboesophageal ganglion (SOe(3)
with oesophagus (Oe) and corpora cardiaca (Cc) emerging from circumoesophageal passage
(COeP). x 1071.
 Internal Anatomy and Morphology of Frankliniella occidentalis 293

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294 DIANE E. ULLMAN
et al.

Fro. 4. Light micrographs showing parasagittal views of structures within thorax, a. Segment of
salivary gland (SG) pressed against a cross section of oesophagus (Oe). × 2375. b. Large portion of
salivary gland (SG). Note large, loosely aggregated cells with dark stained nuclei (n) (arrowhead). ×
2375, c. Cardiac valve (CV) at juncture of oesophagus (Oe) and anterior midgut (Mgl). x 2375.

mesothoracic segment (Figs la, lb, 3, 4a, 4b, 4c). This juncture is delineated by the
cardiac valve (Figs la, 3, 4c). The oesophagus is a flexible structure, apparently
expanding and contracting during the feeding process, as evidenced by the varied
positions in which we observed it (Figs 2, 4a, 4b, 4c). The midgut consists of 3 loops,
designated here for clarity as Mgl, Mg2, and Mg3 (Figs la, lb, 3). While these loops may
be functionally differentiated, we did not observe any morphological differences in this
investigation. The midgut is formed by a single layer of columnar epithelial cells that are
lined with numerous microvilli that extend into the large gut lumen they encircle (Figs
5a, 5b, 6a). Spherocrystals are common in the epithelial cells (Fig. 5c). The cytoplasm
within the villi appears to be oriented along the axis of the microvilli giving the region a
filamentous aspect of fine fibrils running from the villi tip to the cytoplasm immediately
below the point of insertion on the epithelial cells (Figs 6a, 6b, 6c). Two tubular
structures with internal ducts connect the midgut from the junction between Mgl and
Mg2 to the paired salivary reservoirs. The salivary glands and a pair of deferent salivary
canals are then connected to the salivary reservoirs (Figs 3, 7a, 7b). Constrictions in
various parts of the midgut are also observed (Fig. 7c).
The paired salivary glands lie within the prothoracic segment extending posteriorly
from the circumoesophageal passage to the juncture between the pro- and mesothoracic
segments (Figs la, lb). The glands are joined by a narrow piece of tissue and are held in
place posteriorly by fine ligaments that attach to the anterior midgut (Mgl) just below
the cardiac valve (Fig. 3). One gland lies to one side of the oesophagus, while the second
gland is always found on the dorsal surface of the oesophagus (Figs la, lb, 4a, 4b).
 Internal Anatomy and Morphology of Frankliniella occidentalis 295

Fie. 5. TEM micrographs of internal anatomy of midgut, a. Midgut columnar cell (CE) showing
proximity of microvilli (mv) that make up striated or brush border in cross section, large vacuoles (v)
common in these ceils and zonula continua (zc) separating cells. × 11,875. b. Longitudinal view of
microvilli (mv) as they meet midgut columnar cell (CE). × 23,230. c. Midgut columnar cell (CE) in
cross section showing where cell meets midgut lumen (lm), nucleation of cell (n) and spherocrystals
(Sc) commonly found in these cells. × 11,875.
296 DXArqE E. ULLMAN
et aL

FIG. 6. TEM micrographs of midgut microvilli (mv). a. Longitudinal and cross section. Arrowhead
indicates fibrous material within microvilli cytoplasm, x 22,510. b. Higher magnification of midgut
microvilli. Arrowhead indicates fibrous material within microvillar cytoplasm. × 96,190. c. Cross
section of midgut microvilli. Arrowhead indicates fibrous material within microvilli cytoplasm.
× 96,190.
 Internal Anatomy and Morphology of Frankliniella occidentalis 297

FIG. 7. Midgut and ducts from midgut tO salivary gland, a. SEM micrograph showing external aspect
of midgut and juncture where one of 2 ducts (SCMgl) leading from loop in midgut to salivary gland
fastens to midgut, x 875. b. TEM micrograph of cross section of duct shown in Fig. 7a showing
lumen of duct and mitochondria in cellular portion of duct. Arrowhead denotes striated cuticle lining
duct. x 18,760. c. SEM micrograph of midgut. External aspect of columnar cells can be seen. Arrow
denotes typical constriction seen in length of midgut. × 3500.
298 DIANE E. ULLMAN
et aL

The glands are composed of large, loosely aggregated cells that are rich in lipid
droplets, golgi, coated vesicles and numerous membranous lamellar infoldings (Figs 4a,
4b, 8a, 8b). The cells of the salivary glands are further distinguished by large electron
dense or darkly stained nuclei. As already mentioned, 2 ducts lead from the midgut to a
pair of salivary reservoirs (Figs 3, 7a). Each salivary gland has a fine duct extending from
the gland to the reservoir (Fig. 3). At this juncture 2 ducts leave each reservoir (SR1,
SR2). One is common to the duct leaving the gland and also extends into the salivary
reservoir. This duct connects to one arm of a double Y-shaped salivary duct in the
mouthcone that converges into a single canal that in turn empties into the salivarium
(Figs la, 3). The other is a continuation of the duct that enters the reservoir from the
midgut and it connects to the other arm of the Y-shaped salivary duct in the mouthcone
(Fig. 3). The same arrangement is observed for the other salivary gland. Salivary
enzymes or other substances from the midgut may be thus emptied into the salivarium
and may then enter the food canal formed by the maxillary stylets and be injected into
the plant during feeding. No salivary pump or syringe was observed. The mouthcone has
many muscles and pumping may occur by contracting these muscles and constricting the
mouthcone in various ways to force saliva from the salivarium into the plant.
The pyloric valve denotes the juncture between the midgut and the hindgut (Fig. 9).
At this juncture a set of 3 malpighian tubules are found (Figs la, lb, 3, 10, lla, 12a).
Numerous membranous infoldings occur where the malpighian tubules meet the valve
region just anterior to the hindgut forming the basal labyrinth (Fig. 10). Spherocrystals
that are morphologically very similar to those found in the midgut epithelial cells are also
frequently found in the malpighian tubule epithelium (Fig. 10). The malpighian tubules
consist of a single layer of epithelial cells surrounding a lumen and are provided
internally with a brush border of microvilli (Fig. lla). The hindgut has a rich microflora
that appears to be largely rod shaped bacteria (Fig. llb).
The females have a pair of ovaries of which one lies ventral to the midgut and the other
to one side (Figs lb, 12a). Each ovary consists of 4 ovarioles that are suspended
anteriorly from a terminal filament (Figs lb, 12a, 13). Posteriorly, the ovarioles in each
ovary are pressed closely together and merge into a lateral oviduct that eventually
connects to the median oviduct and the vagina (Figs. 12b, 13). Cells within the ovarioles
have large electron dense nuclei (Fig. 13). The ovaries extend the length of the abdomen
and depending upon the stage of ovariole development, they frequently occupy a large
portion of the abdominal cavity. Males have a pair of accessory glands that lie ventral to
the hindgut (Fig. 14). Posterior to the accessory glands are a pair of testes with ducts
leading to a common ejaculatory duct (Fig. 14).

DISCUSSION
Our observations support the concept that certain aspects of Thysanopteran internal
morphology are typical throughout the order, while many other aspects vary widely,
even among species in the same genus. For this reason, some generalizations that can be
made within many other orders of insects are clearly not appropriate when discussing the
Thysanoptera. In the following discussion we present those morphological characteristics
that appear to be general across the order and compare the species that have been
described relative to their differences. Finally, we discuss how some of the morphological
 Internal Anatomy and Morphology of FranklinielIa occidentalis 299

FIG. 8. TEM micrographs showing internal aspects of salivary gland, a. Single cell where it presses
against brain (Br). These ceils are rich in lipids (li) and Golgi apparatus (g). × 9500. b. Juncture
between 2 salivary gland cells showing membranous lamellar infoldings (lmi) common between
salivary gland cells and numerous coated vesicles (cov) (denoted by arrow) that are present
throughout cells, particularly in region of Golgi and lamellar infoldings. 5< 56,400.
300 DIANE E. ULLMAN
et al.

FI6. 9. SEM micrograph of external aspect of pyloric valve where it meets posterior midgut (Mg3).
x 1900. Inset is a light micrograph
showinga section through
pyloricvalve (PV) and its juncture
with
posterior midgut (Mg3) and hindgut (Hg). x 2610.

characteristics we observe in the WFF may influence cellular interactions governing their
ability and efficiency in transmitting TSWV.
The general arrangement of the head, mouthparts and anterior alimentary canal of the
WFT are similar to those of all Thysanopterans that have been studied (Peterson, 1915;
Reyne, 1927; Pesson, 1951; Risler, 1957; Davies, 1958; Mickoleit, 1963; Mound, 1971;
Heming, 1978; Chisholm and Lewis, 1984; Hunter and Ullman, 1989). Our findings
provide support for the conclusions of Peterson (1915), Chisholm and Lewis (1984) and
Hunter and Ullman (1989) that all thrips are equipped with a single mandible, paired
maxillae and a cibarial pump that, in concert act in a piercing-sucking mode and not a
rasping-sucking mode as traditionally thought.
Selection pressures that led to adaptation of the alimentary tract to accommodate
feeding strategies of other piercing-sucking insects, such as the Homoptera, probably
also shaped the feeding structures of the Thysanoptera (Goodchild, 1966; Ullman and
McLean, 1986). Our observations support Peterson's (1915) and Pesson's (1951)
conclusions that there are striking similarities between the Homoptera and the
Thysanoptera in the general arrangement of the head and mouthparts. Like the
Homoptera, the WFT has a precibarial region and preliminary data suggest that the
WFT also have precibarial chemosensilla (Hunter and Ullman, 1989). This region of the
anterior alimentary canal of the WFT differs from that of the Homoptera because it
apparently lacks a precibarial valve at the anterior end of the cibarium (Backus and
McLean, 1982, 1983, 1985; McLean and Kinsey, 1984; Ullman and McLean, 1986).
 Internal Anatomy and Morphology of Frankliniella occidentalis 301

FIG. 10. TEM micrograph of juncture between pyloric valve area and malpighian tubule. Basal
labyrinth (arrowhead) is shown as well as spherocrystals (arrow) that are common in primary cells of
malpighian tubule. Lumens of posterior midgut (Mg3 lm) and hindgut (Hg lm) are shown for
orientation. × 7550.
302 DIANE E. ULLMAN
et al.

FIG. 11. TEM micrographs of malpighian tubule and hindgut, a. Cross section of malpighian tubule
where it presses against hindgut (Hg). Brush border of microvilli (mv) extending into lumen (lm),
numerous rough endoplasmic reticulum (rer), zonula continua (zc) and basal lamina (bsl) are shown.
x 15,010. b. Cross section through hindgut showing numerous rod shaped bacteria found in every
specimen sectioned, x 7125.
 Internal Anatomy and Morphology of Frankliniella occidental& 303

FIG. 12. SEM micrographs of one ovary and oviducts, a. Three of 4 ovarioles pressed against junction
between midgut and hindgut. Ovarioles merge posteriorly into a lateral oviduct (LOvi) (posterior
direction indicated by arrow). One malpighian tubule (MpT) is shown arising from junction between
midgut and hindgut, x 604. b. One of 2 lateral oviducts (LOvi) (posterior direction indicated by
arrow). × 2400.
304 DIANE E. ULLMANet al.

Fie. 13. TEM micrograph of all 4 ovarioles composing one pair of ovaries. Arrows denote
membranes enclosing closely aligned ovarioles. × 11,875. Inset shows electron dense nuclei. × 5700.
 Internal Anatomyand Morphologyof Frankliniella occidentalis 305

FI6. 14. Light micrograph of one pair of testes (Te) where it joins one of relatively
large accessory
glands (AcG). x 2520.

Pesson (1951) depicts a set of muscles encircling the cibarium at its juncture with the
oesophagus in Selenothrips rubrocinctus (Giard), and hypothesizes that these muscles act
as a valve for the cibarium during feeding. We did not observe such muscles, nor are they
depicted in detailed studies of other thrips species (Pesson, 1951; Risler, 1957; Mickoleit,
1963). Therefore, we suggest that in the WFT, closing of the cardiac valve at the juncture
of the oesophagus and anterior midgut must serve 2 purposes: (1) to prevent midgut
contents~from spilling forward during pumping of plant cell contents from the plant into
the anterior alimentary canal, and (2) to create the negative pressure or tension required
to draw cell contents from the plant into the food canal formed by the interlocking of the
maxillary stylets.
Compared to most Homoptera, the oesophagus of the WFT is a relatively large
structure except at the point where it joins the cibarial pump and passes through the very
narrow eircumoesophageal passage (Figs la, lb, 3, 4a). We hypothesize that this
reduction in diameter creates resistance to fluid flow and prevents ingested cell contents
from exiting the stylets as the cibarial pump closes. Instead, the cardiac valve opens and
ingested materials are forced into the much larger oesophagus, and into the midgut. The
relatively large size of the oesophagus probably reflects the WFT's feeding habits that
include ingesting whole chloroplasts from the plant cell cytoplasm (Chisholm and Lewis,
1984). In contrast, the Homopterans that feed primarily on fluids have a much narrower
oesophagus (Ullman and McLean, 1986).
Like all thrips species that have been described the juncture between the oesophagus
and midgut is delineated by the cardiac valve (Peterson, 1915; Sharga, 1933; Pesson,
1951) (Figs la, 4b). The size and length of the oesophagus prior to this junction seems to
vary. In some species, such as Heliothrips hemmorrhiodalis (Bouche), the oesophagus is
extremely slender and extends well into the abdomen before joining the midgut (Pesson,
1951). In contrast, the oesophagus of the WFT is shorter and stouter and joins the
midgut within.the mesothoracic segment.
306 et al.
DIANE E. ULLMAN

All described thrips species have a relatively long, looped midgut. Like Aptinothrips
rufus (Goeze) (Pesson, 1951), the midgut of the WFT is a largely undifferentiated tube.
In many other described species among the Tubilifera and Terebrantia the midgut is
differentiated into three regions (Sharga, 1933; Pesson, 1951). These morphologically
distinct regions may also be functionally differentiated, although this latter question has
never been addressed.
The morphology of the midgut with its single layer of columnar epithelial cells and
internal brush border of microvilli are typical of all members of the Insecta (Figs 5a, 5b).
Kitajima (1975) observed that the cytoplasm within the villi of an unknown species of
Frankliniella was composed of fine fibrous material showing a preferred orientation
along the axis of the microvilli, giving the region a filamentous appearance. We made
similar observations in the midgut microvilli of the WFT and according to Martoja and
Ballan-DuFranqais (1984) this condition is typical of the Insecta. Kitajima (1975) also
observed an unusual glycocalyx on the surface of the microvilli that we did not observe in
the WFT. The exact function of the fine fibrils in the cytoplasm of the villi is unknown.
Kitajima (1975) suggests that they may maintain the rigidity of the microvilli. We did not
observe similar fibrils in the microvillar brush borders present in any of the other internal
organs we investigated (hindgut, malpighian tubules). Therefore, we suggest that the
microfibrils may play an additional role and aid in movement of substrates from within
the midgut to the epithelial cells and eventually the hemolymph.
As in other thrips species described by Pesson (1951) and the Insecta in general, the
pyloric valve delineates the junction between the midgut and the hindgut. The bacterial
organisms we observe in the WFT hindgut are present in all the individuals we have
sectioned regardless of age and/or diet. Therefore, we suggest that they are part of the
insect's normal microflora and may be symbiotic organisms as are known to occur in
many insect species (Houk and Griffiths, 1980). Little is known regarding symbiotic
organisms in the Thysanoptera. Bournier (1961) reported symbiosis for the first time in a
Thysanopteran, specifically Caudothrips buffai (Karny). He observed mycetomes
containing very delicate thread-like bacteria in the oldest ovum of each ovariole and
compares the probable symbiosis to that found in Pediculus. Based on the scant
information in his report, it appears the bacteria he observed are quite different from
those we observe.
The junction between the midgut and the hindgut is also the site at which the
malpighian tubules originate. In contrast to other described thrips species, the WFT has
3, rather than 4 malpighian tubules (Pesson, 1951). The cellular structure of the
malpighian tubules are typical of other insects, with a brush border of tightly packed
microvilli, numerous infoldings of the plasma membrane that extend into the primary
cell, and an abundance of spherocrystals. Mineral spherocrystals are often abundant in
the malpighian tubules of insects (Martoja and Ballan-DuFran~ais, 1984). Midgut and
malpighian tubule concretions have been found in many insects and are thought to
indicate storage of Na, K, Mn, Fe, Zn, Cu, uric acid or represent sequestration of
unusual elements in the diet (Jeantet et al., 1977).
The female reproductive system varies consideraby in shape and size between the
WFT and other described species. In other respects, such as number of ovarioles, all the
studied species are similar. The male reproductive system of the WFT is similar to the
species described by Pesson (1951).
The most unusual and strikingly different anatomical feature we observed in the WFT
 Internal Anatomy and Morphology Frankliniella
of occidentalis 307

is the pair of ducts we observe connecting the midgut and salivary gland (Figs la, lb, 3,
7a, 7b). Pesson (1951) observed a similar pair of structures in Aptinothrips rufus and
describes them as tubular salivary glands, but does not suggest they are ducts. He did not
conduct any histological examination and therefore may have missed seeing the ducts
that were likely within the tubular structures he observed. Although other Frankliniella
species have been studied (Peterson, 1915; Kitajima, 1975), similar structures have not
been reported in other species described.
Our work clearly demonstrates that in the WFT these structures are ducts forming a
union between the midgut and salivary glands. Cells forming the duct are elongated,
nucleated and have an inner lining of striated cuticle. Their connection to the midgut and
the system of ducts leading from the salivary reservoirs out to the mouthcone (Fig. 3),
suggests there may be direct flow of substances between the salivary glands and the
midgut. In addition, it is possible that substances from the midgut may flow to the
salivary reservoirs and exit through the deferent canals (DSC1-DSC4).
Tomato spotted wilt virus is a persistent virus that is thought to be held in the salivary
gland and transmitted during salivation by the insect during feeding (Sakimura, 1960).
The system of salivary ducts we observe hypothetically make it possible for ingested virus
to move directly to the salivary gland from the gut without entering the hemolymph.
Indeed, it may be possible for virus from gut contents to be injected directly into the
salivary secretions via the deferent salivary ducts. These hypotheses warrant further
investigation and may be an important clue to at least one mechanism affecting vector
specificity. By analogy to the Bunyaviridae, to which TSWV bears many similarities,
(Milne and Francki, 1984) it is likely that TSWV replicates in the golgi apparatus and is
transported intracellularly via coated vesicles (DuBois-Dalcq et al., 1984). As Figs 8a and
8b show, the cells composing the salivary gland are rich in these organelles. Although
replication of TSWV in the insect has not yet been demonstrated directly, biological
evidence suggests that virus replication is likely (R. F. L. Mau and J. J. Cho, pers.
comm.). If analogies to the Bunyaviridae continue to hold true, our morphological
studies suggest the salivary glands as a potential replication site.
Striking differences were observed in number, position and ducting of salivary glands,
alimentary canal morphology, and number and arrangement of the malpighian tubules
between the WFT and other studied Thysanopterans. These differences provide support
for the conclusion that internal morphologies may vary widely in this order, perhaps even
between species in the same genus. The results of our investigation support the need for
more detailed morphological studies of other TSWV vector and non-vector thrips
species, particularly to aid in the search for characteristics that may govern vector
specificity and competency.

Acknowledgements--Weare indebted to Gail Murakami, Wes Otani, Laura Gusukuma-Minuto, Mark

Berman, Dr. John J. Cbo, Dr. David Fisher and Dr. Marilyn Dunlap for the technical assistance and facilities
they provided. We also wish to thank Dr. Lynn LeBeck for her thorough review of the article. This research
was supported in part by the United States Department of Agriculture under CSRS Special Grant #88-34135-
3593 managed by the Pacific Basin Advisory Group (PBAG), Biomedical Research Support Grant, University
of Hawaii-Manoa and a Research Centers in Minority Institutions Grant RR-03061, Division of Research
Resources, National Institutes of Health. This is Journal Series no. 3356 of the Hawaii Institute of Tropical
Agriculture and Human Resources.
308 DIANE E. ULLMAN et al.

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APPENDIX

A b b r e v i a t i o nused
s in figures
AcG = A c c e s s o r yG l a n d
b = Bacteria
bl = B a s a lL a b y r i n t h
bsl = B a s a lL a m i n a
Br = Brain
Cb = Cibarium
CbDm = C i b a r i aD
l i l a t o rM u s c l e s
Cc = C o r p o r ac a r d i a c a
CE = C o l u m n a rE p i t h e l i aCells
l
COeP = C i r c u m o e s o p h a g e Pa la s s a g e
Cp = Clypeus
CV = C a r d i a cV a l v e
coy = C o a t e dV e s i c l e s
DSC1, DSC2
DSC3, DSC4 = D e f e r e n tS a l i v a r y C a n a l s
g = Golgi
Hg = Hindgut
lg = Ligaments
li = Lipids
lm = Lumen
lmi = L a m e l l a rI n f o l d i n g s
LOvi = L a t e r a lO v i d u c t
m = Mitochondria
Mc = Mouthcone
310 Diane E. Ullman et al.

Mgl = A n t e r i o r Midgut
Mg2 = Central Midgut
Mg3 = Posterior Midgut
MpT = Malpighian Tubules
mv = Microvilli
n = Nucleus
Oe = Oesophagus
Ov = Ovaries
Ovo = Ovarioles
Pc = Precibarium
PV = Pyloric Valve
rer = Rough Endoplasmic Reticulum
Sal = Salivarium
sc = Spherocrystals
S C M g l , SCMg2 = Salivary Canals Fastened to Midgut
SG = Salivary G l a n d
SG1, SG2 = Paired Salivary Glands
SR1, SR2 = Salivary Reservoirs
SOeG = Suboesophageal Ganglion
Te = Testes
v = Vacuoles
VG = Ventral Ganglia
Y = Y-shaped Salivary Ducts Leading to Salivarium
zc = Zonula Continua