Revista de la Sociedad Entomolgica Argentina

versin On-line ISSN 1851-7471

Rev. Soc. Entomol. Argent. v.66 n.3-4 Mendoza ago./dic. 2007

Larval feeding behavior of the truncatus group of Thrypticus

(Diptera: Dolichopodidae) that breed in the aerenchyma of
Pontederiaceae

Comportamiento alimentario de las larvas del grupo truncatus de Thrypticus

(Diptera: Dolichopodidae) que se cran en el aernquima de Pontederiaceae

Hernndez, M. Cristina

South American Biological Control Laboratory, USDA-ARS. Bolvar 1559, (B1686EFA) Hurlingham,
Buenos Aires, Argentina; e-mail: crisher@speedy.com.ar

RESUMEN. Se da a conocer el comportamiento alimentario de las larvas de algunas especies del

grupo truncatus de Thrypticus (Diptera: Dolichopodidae), que se desarrollan en plantas
hospedadoras de Pontederiaceae. Los estadios inmaduros de T. circularis Bickel & Hernndez se
cran en los pecolos globosos de Eichhornia crassipes (Martius) Solms-Laubach (Pontederiaceae),
minando cerca de la epidermis y abriendo varios orificios al exterior. La larva de T. romus Bickel &
Hernndez crece en E. azurea (Sw.) Kunth, formando una mina curva, corta en comparacin con
las otras especies. La larva de T. formosensis Bickel & Hernndez se desarrolla en Pontederia
cordata L. (Pontederiaceae), excavando la mina entre la epidermis y la gran celda central de los
pecolos. Por ltimo, la larva de T. taragui Bickel & Hernndez se cra en tallos sumergidos de P.
subovata (Seub. in Mart.) Lowden, formando una mina subepidrmica con ramificaciones hacia la
estela central del tallo. No se pudieron asociar las minas correspondientes a las especies T.
yanayacu, T. chanophalus y T. azuricola. No se encontraron predadores ni parsitos de larvas o
pupas, se observaron casos de canibalismo entre larvas cuando el trazado de dos minas confluye.
Las especies del grupo truncatus poseen un modo de alimentacin sumamente especfico,
confinadas al aernquima, se alimentan de la savia extrada de los orificios rodos en los haces
vasculares de los pecolos y tallos, posiblemente con levaduras simbiontes como suplemento para
la nutricin. Numerosas colecciones en el campo y pruebas en el laboratorio, indican que estos
insectos estn asociados con plantas hospedadoras especficas dentro de la misma familia. Esta
especializacin sugiere una larga asociacin insecto-hospedadora.

PALABRAS CLAVE. Grupo truncatus de Thrypticus; Eichhornia crassipes; E. azurea;Pontederia

spp; Fitofagia.
ABSTRACT. Larval feeding behavior of the truncatus group of Thrypticus (Diptera: Dolichopodidae)
that breed in Pontederiaceae species are presented. The larvae of T. circularis Bickel & Hernndez
develop in globous petioles of Eichhornia crassipes (Martius) Solms- Laubach (Pontederiaceae),
digging a mine near the epidermis and forming several holes to the exterior. T. romus Bickel &
Hernndez develop in E. azurea (Sw.) Kunth petioles; the mine is curved and short compared with
other Thrypticus species. T. formosensis Bickel & Hernndez develops in Pontederia cordata L.
(Pontederiaceae), and digs the mine between the epidermis and the big central cell of the petioles.
T. taragui Bickel & Hernndez breeds in submersed stems of P. subovata (Seub. in Mart.) Lowden,
and forms a mine close to the epidermis with branches to the central vascular stele. The mines of T.
yanayacu, T. chanophalus and T. azuricola could not be associated with the corresponding species.
Neither predators nor parasites were found associated with the group, but some cases of
cannibalism were observed when two mines were confluent. The truncatus group species have a
very specific feeding habit, live completely enclosed within host plant tissues, and feed on the sap
obtained from holes chewed in the vascular bundles of the petioles, possibly using yeast as
supplementary nourishment. Extensive field collections and laboratory tests indicate that each
species in the truncatus group is associated with a specific host plant in the Pontederiaceae. Such
specialization suggests a long association between the members of this group and their respective
host plants.

KEY WORDS. Truncatus group of Thrypticus; Eichhornia crassipes; E. azurea;Pontederia spp;

Phytophagy.

INTRODUCTION

The aquatic plant Eichhornia crassipes (Martius) Solms-Laubach

(Pontederiaceae), commonly called waterhyacinth, was released as ornamental in
several countries in the late XIX century. It is currently a mayor invasive weed
throughout the tropics and other regions of the world. It grows very rapidly,
occupying any open water body, covering rivers and lakes, clogging drainage or
irrigation canals. Several methods have been used to control its populations, but to
little avail. Consequently, the search for natural enemies for biological control
commenced in the 1960s (Center, 1994; Cordo, 1999; Julien, 2001). During one of
the first surveys for insects associated with waterhyacinth in Guyana, Surinam and
Brazil, a mining fly larva of Thrypticus Gerstcker was found tunneling the basal
part of the petioles (Bennett & Zwolfer, 1968). Many years later, taxonomic studies
on the Thrypticus associated with Pontederiaceae established that there was a
group of nine new species in South America (Bickel & Hernndez, 2004). Five of
these species, Thrypticus truncatus Bickel & Hernndez (B&H), T. sagittatus B&H,
T. yanayacu B&H, T. chanophalus B&H and T. circularis B&H, develop in
Eichhornia crassipes petioles. Two species, T. romus B&H and T. azuricola B&H,
breed in Eichhornia azurea (Sw) Kunth. Thrypticus formosensis B&H develops in
Pontederia cordata L., and T. taragui B&H breeds in submersed stems of P.
subovata (Seub. in Mart.) Lowden. These nine Thrypticus species each have a
diagnostic oviscapt and male genitalia that allows them to be clearly separated
from congeneric species, and together they form a monophyletic taxon, the
truncatus group.This group was aboundant on E. crassipes and therefore became
the subject of taxonomic and ecological study (Cordo et al. 2000; Bickel &
Hernndez, 2004).
The Pontederiaceae is a small monocotyledonous family composed of six to nine
genera and 35 to 40 species, most of which are native to South America
(Eckenwalder & Barrett, 1986; Barrett & Graham, 1997). All but one of the
Eichhornia species are Neotropical, and all the Pontederia species are New World
(Barrett, 1993). Eichhornia and Pontederia are two major genera in the family, and
there is phylogenetic evidence that Eichhornia is the ancestor of Pontederia.
The petiole anatomy of the leaves of all species in the Pontederiaceae shows
many characters typical of aquatic plants, such as the aerenchyma. This is a
specialized tissue with large cells filled with air that help the plant to float and
exchange gases. In a cross section of the petiole they have a thin layer of
epidermal cells and a central aerenchyma with vascular bundles dispersed
throughout (Castellanos, 1959; Gonzalez, 2002; Mahmood et al., 2005).
Although there are 94 known species of Thrypticus (Grichanov, 2007), their biology
is poorly known (Dyte, 1959; Stiling & Strong, 1983; Dyte, 1993); e.g., the mine of
T. fraterculus (Wheeler) is described as a small blotch or a blister in the leaves of
Scirpus acutus (Muhl.) (Cyperaceae) (Green 1954). All the known Thrypticus
larvae are miners in aquatic or semi-aquatic monocots, and this is the only genus
with phytophagous habits in the Dolichopodidae family (Dyte, 1993).
Both Thrypticus truncatus and T. sagittatus burrow and form mines across the
basal part of a petiole. The larva goes through the aerenchyma finding the vascular
bundles and chewing a small hole in each one, without crossing through them.
They chew through the lateral septa of the aerenchyma until they find other
vascular bundles. In both species, the completed mine has an opening at each
end. The larvae feed on sap from the holes in the bundles, going forward and
backward along the mine. The trace of the larval mines is irregular and it may or
may not be branched, but the mines lack any distinctive pattern diagnostic for
either of the two species. The larva never comes out of its own mine, and it is
incapable of feeding outside of it, and it is unable to dig a new mine if it is removed
and transferred to another part of the aerenchyma. Close to one of the outside
orifices of the mine, the full grown larva will build a chamber where pupation
occurs. The larva cuts an operculum in the epidermis, and afterwards seals the
interior of the chamber with a translucent film, where it rests until pupation with the
body folded in U.
Thrypticus truncatus and T. sagittatus have been studied as biocontrol agents for
waterhyacinth. All the information gathered in that study is presented in another
paper in preparation at this moment (Hernndez, unpubl.).
The objective of this work is to present the larval feeding habits of six species of
truncatus group of Thrypticus which are associated with Eichhornia crassipes, E.
azurea, Pontederia cordata and P. subovata.

MATERIALS AND METHODS

The specimens were collected by H. A. Cordo, A. J. Sosa and the author from 1995
to 2004, during trips in the provinces of Buenos Aires, Entre Ros, Corrientes,
Chaco and Formosa, on the Paraguay-Paran Rivers basins. H. A. Cordo also
surveyed on E. crassipes in the upper Amazon River, near Iquitos in Per (6-22
Nov. 2001), and near Rio de Janeiro, Brazil (5-25 Apr. 2000) (Cordo, unpubl.).
These trips were part of the explorations for the natural enemies of waterhyacinth
performed at the South American Biological Control Laboratory (SABCL), in
Hurlingham, Buenos Aires, Argentina.
Petioles and stems of Eichhornia crassipes, E. azurea, Pontederia cordata and P.
subovata with mines of Thrypticus sp. were gathered. They were kept at room
temperature in opaque bottles with a translucent vial in the cap. The newly
emerged flies were collected from the vials once a day, and preserved in 80 %
ethanol in the SABCL collection. The species identification was carried out by the
author, based on the male genitalia characters. To discover the mine's
characteristics and the larval feeding habits, sections of the petioles were cut and
the mines were traced using a stereomicroscope.

RESULTS

More than 500 adults of Thrypticus truncatus, T. sagittatus, T. circularis, T.

yanayacu and T. chanophalus were obtained from Eichhornia crassipes, about 150
adults of T. romus and 6 adults of T. azuricola from E. azurea, 120 adults of T.
formosensis from Pontederia cordata, and 50 adults of T. taragui from P. subovata.
Each Thrypticus species was obtained invariably from the same host species, this
being evidence of fidelity to host plant in natural conditions.
The length of the larvae depends on the age and the nutrition received, and
therefore the dimensions of the specimens obtained in the field are the result of
several factors. In general, the larvae of these species are 2-3 mm long.
The species of Thrypticus studied in this paper develop inside the aerenchyma of
their host plants. The structure of this aerenchyma is similar between species (Fig.
1), consisting, from the epidermis inwards, of a thin, compact layer of
parenchymatous cells with small vascular bundles right under the epidermis; below
this the petiole is composed by air-filled cells surrounding the vascular bundles
distributed regularly within the petiole. These vascular bundles are bigger than
those found under the epidermis. Particular variations of this basic morphology can
be found among the species.

Figs. 1-9. Thrypticus mines in their host plants. 1, Aerenchyma of Eichhornia crassipes. ac, air-filled
cell;,di, diaphragm; ep, epidermis; sp, septa; vb, vascular bundle. 2, Thrypticus truncatus in E.
crassipes. 3, T. sagittatus in E. crassipes. 4, globous petioles of E. crassipes with T. circularis
mines. 5, detail of T. circularis mines, orifices to exterior. 6, T. romus in E. azurea. 7, T. formosensis
in Pontederia cordata. 8, sub-aquatic rhizome of P. subovata with T. taragui. 9, T. taragui in P.
subovata stem. Arrows in Figs. 2, 4, 6 and 8, signal the mines. Scale bars = 1 cm.
The mines are made by a series of adjacent air cells connected by the holes made
in the septa by the larvae. The size of the holes is always equal to the diameter of
the larval body. There are indications that suggest that when the larva is burrowing
the mine, it may choose the direction of the mine according to the size of the aerial
cell in front of its head. The larva rests its body in the septa of the aerenchyma,
therefore if the size of the airfilled cell immediately ahead is longer than the length
of the first part of the body, it changes direction and moves towards a more closely
positioned septum. Larvae of all the observed species have similar feeding habits,
obtaining their nourishment from the sap of the feeding holes scraped in the
vascular bundles.

Thrypticus species by host plant

Eichhornia crassipes. The two most common species in the Paran-Paraguay

Rivers are Thrypticus truncatus and T. sagittatus. The mines of these species can
have branches of different length and shape (Figs. 2 and 3). In addition, the
species T. yanayacu and T. chanophalus emerged from samples from Per, but
unfortunately the mines were not identified. Thrypticus circularis larvae were
obtained from globular petioles only (Fig. 4). Inside the globular petioles the
aerenchyma has very large cells, and perhaps this is why the mines of this species
are dug near the epidermis, where the cells are smaller, and have several
openings to the exterior (Fig. 5). No larvae of any Thrypticus species were found in
the E. crassipes stems, possibly because it has compact tissues.
Eichhornia azurea: In the E. azurea petioles, the cells of the aerenchyma are
slightly bigger toward the center than near the epidermis. The Thrypticus romus
mines are curved, without ramifications, with two orifices to the exterior, and
comparatively shorter than the mines from other Thrypticus species (Fig. 6). The
Thrypticus azuricola mines have not been identified.
Pontederia cordata: Frequently, the petioles in this plant have a big central cell
along the axis; therefore, the Thrypticus formosensis mines are located between
the epidermis and the central lacuna wall. The mines have some short
ramifications, and may have more than two orifices to the exterior (Fig. 7).
Pontederia subovata: The Thrypticus taragui mines are located in the submerged
stems or in the rhizomes of Pontederia subovata (Fig. 8), but not in the petioles as
in the other species of the group. The stems have a homogeneous aerenchyma
with the vascular bundles concentrated in the central stele and near the epidermis.
Therefore, the main part of the mine is located near the epidermis and has
ramifications to the central stele (Fig.9). The mine has more than two orifices to the
exterior.
All the larvae in the truncatus group are sap feeders but the different species make
a different number of feeding holes. The shortest mines, and consequently the
lowest number of vascular bundles visited, corresponded to Thrypticus romus. The
longest were formed by T. circularis and T. taragui, probably because both species
burrow the mines in the portion of the aerenchyma with smaller vascular bundles.
In addition, it is probable that their nutrition is supplemented by symbiotic yeasts,
given that yeasts were isolated from the larvae of T. truncatus and T. sagittatus
(Hernndez, et al. 2007).
The mortality factors observed were microbial diseases or drowning by mine
flooding. Also, the larvae can attack each other if they are in contact, e.g. when one
mine accidentally intercepts another. Despite the large number of larvae reared,
neither larval nor pupal predators nor parasitoids were found.

DISCUSSION

The truncatus group species share morphological and bionomic characters. They
are all associated to the Eichhornia and Pontederia genera, and use the
microenvironment inside the aerenchyma to develop. This habitat provides them
with nutrition and protection from predators and parasites. Such specialized way of
life associated with two genera of plants, belonging to the same family, can reflect
an ancient insect-host relationship. This assumption is supported by the fact that 7
species in the truncatus group were found on the genus Eichhornia, ancestor of
Pontederia, on which only two species were found.
Thrypticus truncatus, T. sagittatus, T. circularis, T. yanayacu and T. chanophalus,
all of them associated with Eichhornia crassipes, are sympatric in several places
(Bickel & Hernndez, 2004); they use the same environment, the aerenchyma, and
seem to have the same niche, as sap feeders developing enclosed in the
aerenchyma.
Thrypticus circularis has differentiated utilizing only globular petioles, but the other
species live in the same part of the elongated petioles and seem to have similar
behavior. More detailed research is necessary to understand if they really have
overlapped niches, and if they experience inter-specific competition.

ACKNOWLEDGEMENTS

My gratitude to Dr. Axel O. Bachmann for his professional example and unconditional help in
different times during my career. I also want to thank Hugo Cordo and Alejandro Sosa for the
survey's trip; to Guillermo Willie Cabrera, Joaqun Sacco and anonymous reviewers for reading the
manuscript and providing valuable suggestions.

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Recibido: 11-05-2007;
aceptado: 4-09-2007

2011 Sociedad Entomolgica Argentina

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