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Revista de Biología Tropical, ISSN: 2215-2075, Vol. 72: e58265, enero-diciembre 2024 (Publicado Dic. 13, 2024)
Attraction and sexual call in Prodiplosis longifila
(Diptera: Cecidomyiidae): Evidence of a sexual pheromone
Santiago Erazo1, https://orcid.org/0009-0005-3766-2934
Francisco Leiton2 https://orcid.org/0009-0000-7634-7412
Angie Díaz2, https://orcid.org/0009-0003-6561-2454
Valentina Diaz2, https://orcid.org/0009-0004-3982-8216
Juan Fernando Angel2, https://orcid.org/0009-0003-6409-0973
Jan Carlos Achipis2, https://orcid.org/0009-0002-9443-2674
Maria R. Manzano3* https://orcid.org/0000-0002-0858-2426
1. Maestría en Ciencias Agrarias, Universidad Nacional de Colombia sede Palmira, carrera 32 12-00, Palmira, Valle del
Cauca, Colombia; nserazoh@unal.edu.co
2. Ingeniería Agronómica, Universidad Nacional de Colombia sede Palmira, carrera 32 12-00, Palmira, Valle del Cauca,
Colombia; fleiton@unal.edu.co, aldiazl@unal.edu.co, vadiaza@unal.edu.co, jfangelg@unal.edu.co,
jachipis@unal.edu.co
3. Departamento de Ciencias Agrícolas, Universidad Nacional de Colombia sede Palmira, carrera 32 12-00, Palmira, Valle
del Cauca, Colombia; mrmanzanom@unal.edu.co (*Correspondence)
Received 11-I-2024. Corrected 29-IV-2024. Accepted 04-XII-2024.
ABSTRACT
Introduction: Prodiplosis longifila Gagné is a key pest of tomato in Colombia, Ecuador, and Peru. Using a sex
pheromone could be an efficient alternative for its monitoring and control, but its presence is unknown.
Objective: To determine whether virgin females perform sexual calling and whether confined virgin females
attract males.
Methods: In the laboratory, 233 individual virgin females were observed between 5:00-20:30 h using a 60X mag-
nifying glass, and the sexual call was determined by extrusion of the ovipositor. Ten virgin females were confined
in 43 cm3 containers with tulle lids and coated with petroleum jelly to trap males (n = 16). Both containers were
placed equidistantly on the upper inner side of a metal cage with tulle (2 400 cm3) where 10 males were released.
The experiment was repeated with 20 confined females (n = 20). In both experiments, the number of attracted
males was counted 12 hours later. In tomato crops, two Jackson traps were impregnated with odorless glue from
which hung a 43 cm3 plastic container with tulle containing 19-34 virgin females (n = 15). The control containers
had no females. Adult resting sites in the field were searched for.
Results: P. longifila females performed sexual calling during the scotophase and that containers with females
attracted more males than the control. In the field, the attraction was greatest 12 hours after the experiment
was set up. At field P. longifila adults are active during the scotophase and rest during the photophase mainly on
certain plants.
Conclusions: The results indicate that virgin females of P. longifila perform sexual calling and strongly suggest
that females release a sex pheromone.
Key words: sexual behavior; tomato; resting plants; Solanum lycopersicum; key pest.
https://doi.org/10.15517/rev.biol.trop..v72i1.58265
INVERTEBRATE BIOLOGY
2Revista de Biología Tropical, ISSN: 2215-2075 Vol. 72: e58265, enero-diciembre 2024 (Publicado Dic. 13, 2024)
INTRODUCTION
The bud midge Prodiplosis longifila Gagné
(1986) is a key pest of tomato (Solanum lyco-
persicum L.) in Colombia, Ecuador, and Peru.
The adult is small, measuring < 1cm, the female
differs from the male by the shape of its anten-
nae (Gagné, 1994) and because it has a long
ovipositor that remains inside the abdomen
(Gagné, 1986). Larvae feed by sucking the juic-
es of leaf buds, flowers (ovaries and stamens)
and small fruits (Hernandez et al., 2015) and
because of this, the tissues become brown only
after the larvae drop to the soil (Hernandez et
al., 2015). Economic losses of more than 50
% have been estimated (Geraud-Pouey et al.,
2022; Hernandez-Mahecha et al., 2018). The
main and recurrent method of control of P.
longifila is the spraying of synthetic insecticides
without prior knowledge of the population level
of the pest, which has led to its ineffectiveness
(Díaz-Silva, 2011; Duque et al., 2018; Valarezo
et al., 2003) and to the current use of insecticide
mixtures that put human health at risk. In
Colombia, the overuse of insecticides on toma-
toes has led to the production of fruit with
traces of insecticides above the permitted levels
(Bojacá et al., 2013). In addition to the risk to
human health, it is necessary to consider the
rational use of insecticide application because,
among others, biological control agents can
be extremely sensitive to exposure to chemical
synthesis products (Ricupero et al., 2020). In
fact, P. longifila is considered a pest induced
by the high application of insecticides against
other tomato pests which most likely negatively
affected its biological control (Geraud-Pouey et
al., 2022) and parasitoid wasps such as Synopeas
varipes Harrignton 1900 and Synopeas reticu-
latifrons Buhl 2011 only appear at the end of
the crop cycle when farmers stop insecticide
sprays (Hernandez-Mahecha et al., 2018). It is
necessary to develop more specific monitoring
and control alternatives for this Cecidomyiid,
such as the use of sex pheromones. Precisely,
the chemical composition of sex pheromones
RESUMEN
Atracción y llamado sexual en Prodiplosis longifila (Diptera: Cecidomyiidae):
Evidencia de una feromona sexual
Introducción: Prodiplosis longifila Gagné es una plaga clave del tomate en Colombia, Ecuador y Perú. El uso
de una feromona sexual podría ser una alternativa eficiente para su monitoreo y control, pero se desconoce su
presencia.
Objetivo: Determinar si las hembras vírgenes realizan llamados sexuales y si las hembras vírgenes confinadas
atraen a los machos.
Métodos: En el laboratorio se observaron 233 hembras vírgenes individualizadas entre las 5:00-20:30 h utilizando
una lupa de 60X y se determinó el llamado sexual por extrusión del ovipositor. Diez hembras vírgenes fueron
confinadas en recipientes de 43 cm3 con tapas de tul y recubiertos de vaselina para atrapar machos (n = 16).
Ambos recipientes se colocaron equidistantes en la parte superior interna de una jaula metálica con tul (2 400
cm3) donde se liberaron 10 machos. El experimento se repitió con 20 hembras confinadas (n = 20). En ambos
experimentos, el número de machos atraídos se contó 12 horas después. En cultivos de tomate, se impregnaron
dos trampas Jackson con un pegante inodoro de las que colgaba un recipiente de plástico de 43 cm3 con tul que
contenía 19-34 hembras vírgenes (n = 15). Los recipientes control no contenían hembras. Se buscaron sitios de
reposo de los adultos en campo.
Resultados: Las hembras de P. longifila realizaban llamados sexuales durante la escotofase y los recipientes con
hembras atraían a más machos que el control. En el campo, la atracción fue mayor 12 horas después del inicio
del experimento. En campo, los adultos son activos durante la escotofase y descansan durante la fotofase princi-
palmente en ciertas plantas.
Conclusiones: Los resultados indican que las hembras vírgenes de P. longifila hacen llamado sexual y sugieren
fuertemente que las hembras liberan una feromona sexual.
Palabras clave: comportamiento sexual; tomate; plantas de reposo; Solanum lycopersicum; plaga clave.
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has been studied in at least 17 species of cecido-
mids, some of which are already commercially
produced and used to monitor populations of
pest species (Hall et al., 2012; Xu et al., 2020).
Among the cecidomid species monitored with
sex pheromones are Contarinia nasturtii (Kief-
fer, 1888) (Hillbur et al., 2005), Dasineura mali
(Kieffer, 1904) (Cross & Hall, 2007; Liu et al.,
2009), Dasineura oxycoccana (Johnson, 1899)
(Fitzpatrick et al., 2013), Dasineura plicatrix
(Loew, 1850) and Mayetiola destructor (Say,
1817) (Knutson et al., 2017). The emission of
sex pheromones is associated with morpho-
logical and behavioral characteristics of the
cecidomid. Although the presence of a sex
pheromone in P. longifila is unknown, two
morphological characteristics suggest it. First,
the presence of sensoria called circumfila that
girdle the flagellomeres and form loops on the
antenna of males (Hernandez-Mahecha et al.,
2018; Gagné, 1994), and second the presence
of a long telescopic ovipositor that is typical of
insects that reproduce with emission of a pher-
omone (Hall et al., 2012; Van Lenteren et al.,
2002; Zhang et al., 2021). Adults are only active
in the dark and during the light hours rest on
some plants (Díaz-Silva, 2011). Based on these
morphological characteristics and behavior
suggesting the importance of chemical com-
munication mediated by a sex pheromone for
mating, the objective of this research was to
establish whether P. longifila females performed
sex calling to attract males and to determine
whether virgin females confined in laboratory
and field conditions attracted males. Adult rest-
ing places were determined.
MATERIAL AND METHODS
Insects: Leaflets of Solanum lycopersicum
L. tomato with P. longifila larvae were col-
lected from commercial crops located in several
localities of Valle del Cauca, Colombia (Pradera
3°25”9’ N, 76°14”34’ W; Rozo 3°36”53’ N,
76°23”10’ W; Santa Elena 3°39”25’ N, 76°14”21’
W, Trujillo 4°12”41’ N, 76°19”13’ W). Larvae
were transported in plastic boxes with wet
paper to the Entomology laboratory of the
Universidad Nacional de Colombia sede Palmi-
ra (3°30”42’ N, 76°18”28’ W) and maintained
at average conditions of 24.1 °C ± 0.36 SE, 72.9
% RH ± 0.7 SE and photoperiod of 12L:12D to
continue their development.
Pupae were maintained in moistened clay
soil according to the method suggested by
Geraud-Pouey et al., (2022). Soil was obtained
from an abandoned cacao (Theobroma cacao
L.) crop that was dried for 48 h at 200 ˚C and
sifted with a 60-inch sieve to obtain a fine sub-
strate. The emerged adults were continuously
sexed soon after emergence to avoid mating
according to the type of antennae and genitalia
(Gagné, 1986; Gagné, 1994) through a Nikon
SMZ 745 stereoscope and red light produced
by three 60 W glass bulbs painted red so as not
to disturb the insects. Although P. longifila is
the only species of Cecidomyiidae present on
S. lycopersicum in Colombia (Velasco-Cuervo
et al., 2016), 5 adults/night were routinely sam-
pled to corroborate species identity according
to taxonomic keys (Gagné, 1986; Gagné, 1994).
Laboratory experiments were conducted
in the Entomology Laboratory and field experi-
ments were conducted in a commercial tomato
crop during flowering and fructification, locat-
ed in the village of Cedrales, municipality of
Trujillo, Valle del Cauca, Colombia (4°13’35” N,
76°19’24” W) at 1 596 m.a.s.l., annual average of
21.5 °C and 78.1 % RH.
Calling behavior: To determine the pres-
ence and time of the sexual call, 233 newly
emerged virgin females were placed in indi-
vidual in 1.5 oz plastic cups and observed every
30 minutes between 5:00-20:30 h. To avoid
disturbing the insects, observations were made
with 60X Walfront magnifying glass with LED
light. Calling females extruded partially the
ovipositor (only the final part of the oviposi-
tor is shown accompanied by contractions and
circular movements of the ovipositor) or totally
(ovipositor completely visible). Each observa-
tion by using LED lights lasted 20 seconds
because previous observations determined that
this lasts from 10 to 15 seconds. Females were
observed until death. The number of females
4Revista de Biología Tropical, ISSN: 2215-2075 Vol. 72: e58265, enero-diciembre 2024 (Publicado Dic. 13, 2024)
extruding the ovipositor (partial or total) at
each observation time was plotted.
Attraction of males to confined females
of Prodiplosis longifila under laboratory con-
ditions: The experiments were conducted with
males and females of P. longifila emerged on the
same day. The adults emerged from 15:00 h and
around 18:00 h the required number of females
and males for the experiment was completed.
The experiment started between 19:00-20:00 h.
Virgin females were confined in a 43 cm3
plastic container with a lid lined with synthetic
tulle mesh allowing air flow and covered on
the outside with petroleum jelly to trap attract-
ed males (treatment); a similar container, but
without females, was the experimental control.
A control and a treatment container arranged
upside down were velcroed equidistantly to the
inside top face of a tulle-lined aluminum cage
(10 cm length x 12 cm width x 20 cm height).
Inside the cage, groups of newly emerged vir-
gin males were released and had free choice to
fly to the treatment or the control. In the first
group of experiments (n = 16) 20 females were
confined and 10 males were released (ratio 1
male: 2 females). In the second group of experi-
ments (n = 20), 10 females were confined, and
10 males were released (ratio 1 male: 1 female).
Both males and females were used only once.
During the experiment, the cages were kept
inside the laboratory covered with a black cloth
to reduce the influence of any artificial light.
Twelve hours later, the number of males attract-
ed and attached to the treatment and control
containers were counted. The treatment and
control containers were used only once, and
the aluminum cages were washed with water
and cleaned with alcohol between experiments.
Attraction of males to confined females
of Prodiplosis longifila under field conditions:
The experiment was conducted on a commer-
cial crop of tomato S. lycopersicum Libertador
variety planted in a semi-covered area of 1.3
ha with distances of 0.40 m between plants and
1.4 m between rows with approximately 21000
plants in the phenological stage of flowering
and fruiting. Two Jackson traps (n = 15) made
of cardboard (SÁFER®) were installed in one
furrow of the crop at a height of 1 m from
the ground, which corresponds approximately
to the middle part of the plants, separated by
2m, and protected from the rain with a zinc
roof covered with PVC. In the center of each
trap, a 43 cm3 plastic container with a lid lined
with synthetic tulle mesh to allow air flow was
placed face up. Between 19-34 newly emerged
virgin P. longifila females were confined in the
treatment Jackson trap container. The con-
trol trap container was empty. The confined
females were from the biological development
of larvae collected in the experimental culture.
All surfaces of the Jackson traps and jars were
impregnated with Sáfertac® (SÁFER) odorless
glue to trap attracted males. Confined females
were fed by offering the insects balls of cotton
soaked in 10 % sugar solution to prolong their
longevity. Treatment and control traps were
installed between 18:00-19:00 hours and 12, 24
and 36 hours after installation the number of
males attracted to each trap was counted with
the aid of a 60X magnifying glass (Warrant).
Each group of P. longifila females was used only
once (n= 15). Captured males were removed
after each evaluation.
To know the population density of P. l o ng i -
fila in the crop that would guarantee the pres-
ence of males in the crop, the larval population
of P. longifila was sampled and counted weekly
based on what was suggested by Duque et al.,
(2018). Three crop furrows were randomly
selected, and 10 leaf buds were randomly taken
from each furrow (30 leaf buds/week/crop).
Using a 60X magnifying glass, the number of
longifila larvae was counted and the average
number of larvae per leaf shoot in the crop
was determined. Since the development time
between the first instar larva and the adult P.
longifila in tomato is 14 days (Duque et al.,
2018), continuous emergence of adults in the
crop was expected. Additionally, 15 nocturnal
visits were made to the crop to observe and
confirm the presence of adults, and five indi-
viduals were randomly collected/night to con-
firm their taxonomic identity in the laboratory.
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Resting sites of Prodiplosis longifila
adults in the field: Daytime (between 8:00-
17:00 h) and nighttime (between 18:00-19:00 h)
searches for adults of P. longifila were conducted
on uncultivated plants growing spontaneously
in the surroundings of the tomato crop. Flash-
lights with LED light were used at night. The
plants with adults were pressed and transferred
to the Josep Cuatrecasas y Arumí Herbarium
of the Universidad Nacional de Colombia sede
Palmira for taxonomic identification.
Statistical analysis: The number of males
was expressed as mean ± standard error (SE).
For the analysis of attraction in the labo-
ratory, the Analysis of Variance (ANOVA)
analysis indicated a significative effect of the
treatment (presence/absence of females) (F =
48.15, df (1,68), p < 0.05). Using the residual
values of ANOVA there were tested normal-
ity and homogeneity of variances assumptions.
Normality distribution was indicated by the
Anderson-Darling test (AD = 0.617, p = 0.105).
Homogeneity of variances was tested with
Bartlett’s test (Barlett´s test statistic = 11.24, p
= 0.010) (SAS 9.2, 2008). Because the variances
were not homogeneous and the values from
numerical counts of attracted males were very
small, the variable number of males was trans-
formed with √(x+0.5). With the transformed
data, Levene’s test confirmed the assumption
of equal variances. For attraction experiments
under laboratory conditions the number of
males attracted between control and treat-
ment, and between female: male ratios of 1:1
and 2:1 and their interaction were compared
through an ANOVA. Means were compared
with Tukey’s test p < 0.05. For experiments at
field conditions the number of males attracted
between control and treatment at 12 h, 24 h
and 36 h was compared. The assumption of
homogeneity of variances was not met for the
24 h male field attraction experiments (F =
0.10, p < 0.05) but the assumption of normal-
ity was met (Ryan Joiner test, RJ = 0.976, p >
0.100), therefore Welch’s non-parametric test
and Games-Howell mean comparison test with
p <0.05 were used. No normality at 24 h and
36h was indicated by the Ryan Joiner test (RJ
= 0.908, p <0.010 and RJ = 0.885, p < 0.010
respectively). The assumption of homogene-
ity of variances was met for the 24h male field
attraction experiments (Levene test, p = 0.134);
for the 36 h experiment it was proposed homo-
geneity of variances because the control was
cero (no insects were attracted to the females).
Number of males attracted between control
and treatment at 24 h and 36 h were compared
using one-way ANOVA. Means were compared
with Tukey’s test p < 0.05.
RESULTS
Calling behavior: The emergence of adults
occurred after 15:00 h. Adults were not highly
active for about 20 min after emergence, which
allowed them to be sexed more easily. Females
of P. longifila performed sexual calling through
complete (dark bar) or partial extrusion (pale
bar) of the ovipositor mainly in the early morn-
ing (5:00-7:30 h), in the afternoon (16:30-18:00
h) and in the evening (18:30-20:30 h) (Fig. 1).
There was a higher frequency of complete ovi-
positor extrusion (387 times) versus partial ovi-
positor extrusion (242 times) (Fig. 1). Females
showed the ovipositor at least once up to 35
hours after emergence, however, after this time
and up to 96 hours of follow-up (day 4) no ovi-
positor extrusion was evident. It was found that
95 % of the females evaluated showed complete
or partial ovipositor for the first time during the
first 4 hours after emergence. Ovipositor extru-
sion occurred mainly during scotophase.
Attraction of males to confined females
of Prodiplosis longifila under laboratory con-
ditions: A higher average number of P. longifila
males were attracted to the container with vir-
gin females than to the control in the 20 females
:10 males densities (Fig. 2).
Similarly in the 10 females:10 males densi-
ties more males were attracted to the treatment
than to the control (Fig. 3). The number of
males attracted was significantly different from
the control for both ratios (ANOVA, F = 47.82,
df = 1.68, p < 0.05; Tukey p < 0.05). However,
6Revista de Biología Tropical, ISSN: 2215-2075 Vol. 72: e58265, enero-diciembre 2024 (Publicado Dic. 13, 2024)
comparison between the attraction proportions
showed no significant difference in the number
of males attracted (ANOVA, F = 0.00, df = 1.1,
p = 0.959; Tukey p > 0.05).
Attraction of males to confined females
of Prodiplosis longifila under field condi-
tions: Field monitoring showed an average
population of 12.5 ± 1.3 larvae/ leaf shoot in
the tomato lot during the experiment. Twelve
hours after confinement of the females, a sig-
nificant difference was found between the treat-
ment and the control (Welch’s test, F = 9.46 df
= 1.16.89, p = 0.007; Games-Howell p < 0.05),
but no significant difference was found at 24 h
(ANOVA, F = 2.38, df = 1.28, p = 0.134; Tukey p
> 0.05) or at 36 h (ANOVA, F = 3.50, df = 1.28,
p = 0.959; Tukey p > 0.072) (Fig. 4).
Fig. 1. Number of P. longifila females (n = 233) with complete and partial extrusion of the ovipositor during four days since
emergence. The number above the bar indicates the number of females that performed these behavioral events.
Fig. 2. Average number of P. longifila males attracted to
confined females under laboratory conditions (ratio of
attraction of 20 females to 10 males, n = 16), different letters
indicate significant differences (Tukey test, p < 0.05).
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Resting sites of Prodiplosis longifila
adults in the field: No flight activity of P. l o n-
gifila adults was observed during photophase
between 7:00 and 16:00 hours; after 16:00 hours
adults moved with rapid movement around
tomato plants and between the crop furrows.
Only adults (not larvae) were found resting
on the foliage of spontaneously low-growing
plants identified as Wedelia sp. and Spilanthes
acmella Murr, both from the Asteraceae family.
Adults were also found in areas with humid and
shaded microclimates, for example, under the
guadua (Guaduaangustifolia Kunth) structures
where tomato fruits are left to be packed and
marketed, and under an abandoned guadua
support that was outside the crop where the
two species of Asteraceae named above were
growing in the middle of pastures. During sco-
tophase the adults remained still and visible on
leaf buds and on the upper and lower sides of
tomato leaves.
DISCUSSION
The results of the sexual call of P. longifila
coincide with those reported for other ceci-
domid species that release a sex pheromone.
Virgin females of P. longifila perform sexual
calling through total or partial extrusion of the
ovipositor accompanied by circular movements
of the ovipositor. This has been observed in
other cecidomid species that through the exten-
sion of the ovipositor emit sex pheromones that
are produced by specialized glands present in
Fig. 3. Average number of P. longifila males attracted to
confined females under laboratory conditions (ratio of
attraction of 10 females to 10 males, n= 20), different letters
indicate significant differences (Tukey test, p < 0.05).
Fig. 4. Number of P. longifila males attracted to field-confined females and the control 12, 24 and 36 hours after installation
(n = 15). Different letters show differences between treatment and control (for 12 h, Games-Howell post Welch’s test p < 0.05;
for 24 h and 36 h, Tukey post one-way ANOVA p > 0.05).
8Revista de Biología Tropical, ISSN: 2215-2075 Vol. 72: e58265, enero-diciembre 2024 (Publicado Dic. 13, 2024)
their abdomen (Boddum et al., 2010; Hall et al.,
2012; Xu et al., 2020) and that with movements
and postures facilitate the dispersion and distri-
bution of the pheromones to the exterior (Cade,
1985). For example, virgin females of Contarin-
ia oregonensis Foote immediately prior to mat-
ing “call” males by spreading and shaking back
and forth their ovipositors (Miller & Borden,
1984). During sexual calling the extended ovi-
positor of Dasineura brassicae Winn can even
double the length of the female’s body (Isidoro
et al., 1992) and these telescopic ovipositors
such as the one of P. longifila are characteristic
of females emitting sex pheromones (Riolo et
al., 2014). It is not clear if pheromone could be
released during the partially ovipositor extru-
sion observed on P. longifila. Emergence of P.
longifila after 16:00 h and sex calling during
the hours of darkness has also been observed
in cecidomids that produce a sex pheromone
such as Sitodiplosis mosellana Géhin (Bruce
et al., 2007; Gries et al., 2000). Their females,
after emerging in the late afternoon begin to
extend their ovipositors, increase sexual calling
gradually during the last hours of photophase
but call more frequently in scotophase (Pivnick
& Labbé, 1992). Prodiplosis longifila females
called at least once during the first four hours
after emergence (95 % of females) suggesting
that due to their short longevity (1.1 days ±
0.05) (Duque et al., 2018) their sexual maturity
is rapid and mating must occur early, adaptive
characteristics of insects with r-reproductive
strategy (Pianka, 1982). This behavior has also
been reported in females of M. destructor,
which due to their short longevity (Harris
& Rose, 1991), immediately after emergence
mature oocytes and in the first four hours of the
morning extend their ovipositor to emit a sex
pheromone that attracts males for copulation
(McKay & Hatchett, 1984).
The increased activity of adults of P. l o n -
gifila during the scotophase coincides with
our nocturnal observations made in the field
which showed that during the day adults are
inactive and take refuge in plants or guadua
structures but are active during the scotophase.
Valarezo et al., (2003) reported that copulation
of P. longifila occurs during the night and Díaz-
Silva (2011) indicated that adults are active dur-
ing the night. The presence of P. longifila adults
on plants such as Wedelia sp. (Asteraceae) and
Spilanthes acmella Murr (Asteraceae) opens the
possibility of investigating them as trap plants
within an integrated management program
for P. longifila in the tomato agroecosystem.
Delgado (1998) reported the presence of P. l o n -
gifila larvae in the Urticaceae Fleurya aestuans
L. while, in Peru, Díaz-Silva (2011) reports P.
longifila larvae in two other plant species of the
Asteraceae family which are Cynara scolymus
L. (damage on shoots of seedlings or tender
plants) and Tagetes erecta L. (scraping on flower
buds). It is unknown whether P. longifila takes
nectar from these plant species, but adult lon-
gevity is known to be increased by consuming
sugar (Duque et al., 2018) and that after flower-
ing of the tomato crop P. longifila infestation
increases suggesting the importance of flowers
in the ecology of the insect.
Attraction of males to confined females
of Prodiplosis longifila: Confined virgin
females of P. longifila exerted attraction behav-
ior on virgin males most likely mediated by the
production of a sex pheromone. Our experi-
ments took place during the scotophase, a
period that included the sexual calling phase.
Insects in general, including moths, are gov-
erned by a circadian clock that allows females
to synchronously release pheromones, which
in turn will generate an attraction response
in males during a set time (Gadenne et al.,
2016; Groot, 2014). For P. longifila, pheromone
release and male attraction at laboratory con-
ditions was tested during a 12 h period using
adults up to three hours of emergence. Density
variation of confined P. longifila females had
no significant effect on the number of attracted
males. This could be explained on the one hand,
because Cecidomyiids have long circumfila
sensoria (Zhang et al., 2021) present in P. l o n g i -
fila (Gagné, 1994; Hernández et al., 2015) and
which structurally possess abundant dendritic
neuronal branches around each antennal fla-
gellum node, indicating an olfactory function
9
Revista de Biología Tropical, ISSN: 2215-2075, Vol. 72: e58265, enero-diciembre 2024 (Publicado Dic. 13, 2024)
and a close relationship with female sex phero-
mone detection (Boddum et al., 2010; Crook
& Mordue 1999; Hillbur et al., 2001; Solinas &
Nuzzaci, 1987). On the other hand, cecidomid
pheromones are highly selective, potent, and
efficient (Witzgall et al., 2010), which added to
the fact that our attraction experiments under
laboratory conditions were conducted at a short
distance (20 cm) would favor attraction of
males with at least 10 virgin females. For exam-
ple, in a short distance wind tunnel experiment
(horizontal plane ranging from 10 to 35 cm), M.
destructor males were exposed to 20 ng and 2 ng
of their sex pheromone compound (2S)-(E)10-
tridecen-2-yl acetate and both doses elicited
a significant response in the flight attraction
response of males to the compound (Harris &
Foster, 1999). Field experiments on the ceci-
domid Haplodiplosis marginata (von Roser),
where pheromone dispensers were loaded with
the compound nonan-2-yl butanoate at doses
of 5 mg and 10 mg, showed that both doses
generated a male attraction response (Censier
et al., 2016).
Field experiments indicate that a greater
number of males were attracted 12 hours after
setting up the experiment. During this time,
virgin females attracted 57.9 % of the males
attracted during the entire experiment, a trend
that was similar in the laboratory trials where
on average 56.7 % of males flew to females. In
this regard, Hodgdon et al., (2019) found that
in Brassicaceae crops, 73 % of C. nasturii males
were caught in Jackson traps with commercial
pheromone during the first 5 hours after emer-
gence and Isidoro et al., (1992) reported that
57 % of virgin D. brassiccae females performed
sexual calling on the first day of life. In the
experiments of Hodgdon et al., (2019), from 5 h
to 24 h after emergence, the attraction of males
decreased significantly, similar to what was
found in P. longifila in the field, because after 12
h the attraction of virgin females decreased; this
result matches our sex call experiment because
between 16 h and 35 h after the emergence of P.
longifila the frequency of sex calling decreased
until it disappeared after 35 h after the emer-
gence of adults.
The fact that P. longifila females in the
field attracted mostly males during a relatively
brief time span and at laboratory conditions 95
% of the females showed complete or partial
ovipositor for the first time during the first 4
hours after emergence suggest several explana-
tions. First, adaptation to rapid reproduction in
short-lived insects such as P. longifila (Duque et
al., 2018), second, a strong and reliable attrac-
tion to the emitted infochemical (Witzgall et
al., 2010) and third that females move soon
after emergence. In this regard, Valarezo et al.,
(2003) mention that the insect pupae fall to
the ground or adhere to the branches, stems,
or leaves of the tomato and that copulation
can occur on the same plant where the females
emerge and perch so that the males initiate a
courtship flight by vividly flapping their wings.
For C. nasurtii it is suggested that the insect
mates near emergence sites because females
extended their ovipositors when emerging from
the ground (Hodgdon et al., 2019).
The results presented strongly suggest that
females of P. longifila emit a sex pheromone in
glands located in the abdomen and that they
produce an attraction response at short dis-
tance under laboratory conditions and at long
distance under field conditions. It is necessary
to detect and identify this pheromone through
molecular and chemical techniques to develop
an attractant for monitoring and control of this
pest in tomato.
Ethical statement: the authors declare that
they all agree with this publication and made
significant contributions; that there is no con-
flict of interest of any kind; and that we fol-
lowed all pertinent ethical and legal procedures
and requirements. All financial sources are fully
and clearly stated in the acknowledgments sec-
tion. A signed document has been filed in the
journal archives.
ACKNOWLEDGMENTS
The authors would like to thank Universi-
dad Nacional de Colombia for research support
(Hermes 49236 and Hermes 54385), to Paola
10 Revista de Biología Tropical, ISSN: 2215-2075 Vol. 72: e58265, enero-diciembre 2024 (Publicado Dic. 13, 2024)
Rodriguez (Bayer Colombia), and tomato farm-
ers for logistical support during insect sam-
pling; to Herbario Josep Cuatrecasas y Arumí
at the Universidad Nacional de Colombia sede
Palmira for identifying the plants and to Nor-
bey Marin for statistical analysis support.
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