Molecular and morphological diagnoses of five species of Trichogramma: Biological control agents of Chrysodeixis chalcites (Lepidoptera: Noctuidae) and Tuta absoluta (Lepidoptera: Gelechiidae) in the Canary Islands

Abstract

Prospecting for potential natural enemies of the invasive lepidopteran tomato pest Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) and the banana pest Chrysodeixis chalcites (Esper) (Lepidoptera: Noctuidae) on the Canary Islands archipelago, where no Trichogramma species were previously recorded, has led to the discovery of five distinct species. T. achaeae Nagaraja & Nagarkatti, T. bourarachae Pintureau & Babault, T. euproctidis (Girault) and T. evanescens Westwood are relatively widespread species. The fifth is close to T. brassicae Bezdenko, but differs sufficiently in the sequence of the ITS2 region of ribosomal RNA to warrant further investigation as
a species probably new to science. Each species is treated in detail in order to facilitate identification in future using
molecular and/or morphological characters, or a combination of both. All species are newly recorded for the Canary Islands,
and the distribution of each within the islands and elsewhere is provided. Known host records are given within the Canary
Islands and elsewhere. The most common species found, T. achaeae, is already being used in biological control programmes against T. absoluta in mainland Spain and field trials are ongoing to evaluate its effectiveness as a biological control agent of C. chalcites in banana crops.

Full text

Molecular and morphological diagnoses of five species
of Trichogramma: biological control agents of Chrysodeixis
chalcites (Lepidoptera: Noctuidae) and Tuta absoluta
(Lepidoptera: Gelechiidae) in the Canary Islands
Andrew Polaszek •Paul F. Rugman-Jones •
Richard Stouthamer •Estrella Hernandez-Suarez •
Toma
´s Cabello •Modesto del Pino Pe
´rez
Received: 1 September 2010 / Accepted: 15 April 2011 / Published online: 12 May 2011
ÓInternational Organization for Biological Control (IOBC) 2011
Abstract Prospecting for potential natural enemies of
the invasive lepidopteran tomato pest Tuta absoluta
(Meyrick) (Lepidoptera: Gelechiidae) and the banana
pest Chrysodeixis chalcites (Esper) (Lepidoptera: Noc-
tuidae) on the Canary Islands archipelago, where no
Trichogramma species were previously recorded, has
led to the discovery of five distinct species. T. achaeae
Nagaraja & Nagarkatti, T. bourarachae Pintureau &
Babault, T. euproctidis (Girault) and T. evanescens
Westwood are relatively widespread species. The fifth is
close to T. brassicae Bezdenko, but differs sufficiently
in the sequence of the ITS2 region of ribosomal RNA to
warrant further investigation as a species probably new
to science. Each species is treated in detail in order to
facilitate identification in future using molecular and/or
morphological characters, or a combination of both. All
species are newly recorded for the Canary Islands, and
the distribution of each within the islands and elsewhere
is provided. Known host records are given within the
Canary Islands and elsewhere. The most common
species found, T. achaeae, is already being used in
biological control programmes against T. absoluta in
mainland Spain and field trials are ongoing to evaluate
its effectiveness as a biological control agent of
C. chalcites in banana crops.
Keywords Trichogramma achaeae 
T. bourarachae T. evanescens T. euproctidis 
Tuta absoluta Chrysodeixis chalcites Musa
acuminata Solanum lycopersicum Egg parasitoid 
Ooparasitoid Invasive pest Canary Islands
Introduction
The South American tomato pinworm Tuta absoluta
(Meyrick) (Lepidoptera: Gelechiidae) is a major pest
of tomato and other solanaceous crops that has spread
from Central America to most of South America
(EPPO 2005) and was accidentally introduced into
Spain in 2006 (Urbaneja et al. 2007; EPPO 2008) and
Handling Editor: Dirk Babendreier
A. Polaszek (&)
Department of Entomology, Natural History Museum,
London SW7 5BD, UK
e-mail: ap@nhm.ac.uk
P. F. Rugman-Jones R. Stouthamer
Entomology, University of California,
Riverside, CA 92521, USA
E. Hernandez-Suarez M. del Pino Pe
´rez
Department of Proteccio
´n Vegetal, Instituto Canario de
Investigaciones Agrarias, P.B. 60, 38200 La Laguna,
Tenerife, Islas Canarias, Spain
T. Cabello
Centro de Investigacio
´n en Biotecnologı
´a
Agroalimentaria, Universidad de Almerı
´a,
Ctra Sacramento s/n, 04120 Almerı
´a, Spain
123
BioControl (2012) 57:21–35
DOI 10.1007/s10526-011-9361-y

subsequently into other European countries. It is
regarded as an established pest in Spain (EPPO 2008)
much of Southern Europe and North Africa, and is
rapidly spreading east across the Mediterranean
(FERA 2010; Roditakis et al. 2010) and into Northern
Europe.
Tuta absoluta is extremely difficult to control
using chemical insecticides because larvae mine
within plant tissue and are thus protected at least
from contact insecticides (Branco and Franca 1993),
but also because of the development of resistance
(Lietti et al. 2005; Siqueira et al. 2000). For these
reasons biocontrol using parasitoids has been inves-
tigated on many occasions as a potentially suitable
control method both in Latin America (Bueno 2005),
including the use of Trichogramma species there
(Domingues et al. 2003; Faria et al. 2008; Freitas
et al. 1994; Villas-Boas and Franca 1996) and more
recently in Europe (Cabello et al. 2009b). Although
several species of autochthonous predators have been
evaluated as biological control agents in Spain,
particularly the mirid bugs Nesidiocoris tenuis
(Reuter) (Urbaneja et al. 2008) and the damsel bug
Nabis pseudoferus Remane (Cabello et al. 2009a) the
establishment into the crops is often too slow to avoid
damage to the crop, and consequently chemical
treatments are needed.
The golden twin-spot moth or tomato looper,
Chrysodeixis chalcites (Esper) (Lepidoptera: Noctui-
dae) is one of the most damaging pests of banana
crops (Musa acuminata) in the Canary Islands,
inflicting serious economic damage to the youngest
leafpipes, leaves and fruits (Perera and Molina 2007;
Cabello 2009; del Pino et al. 2011). It is a subtropical
species, occurring in Africa, Oceania, Southern
Europe and South Asia, attacking a large number of
cultivated plants (Cayrol 1972). In Southern Spain
C. chalcites has had an important economic impact in
the Guadalquivir Valley (Cabello 1986), la Vega of
Granada (Cabello 1988) and in greenhouses in the
province of Almerı
´a (Cabello et al. 1996). It is
considered a major pest in Israel and Egypt (Harakly
and Farag 1975), Italy (Zandigiacomo 1990), Bul-
garia and Turkey (Loginova 1992; Uygun and Ozgur
1980), The Netherlands (Vos and Rutten 1995) and
Belgium (Veire 1993).
Control of C. chalcites has failed for reasons
similar to those for the failure of T. absoluta control.
C. chalcites adults deposit eggs individually (Cayrol
1972) or in small groups (Harakly and Farag 1975) and
always highly dispersed in the crop. The inefficiency of
applied synthetic and biological insecticides has been
demonstrated (Alonso 2009; del Pino et al. 2011), and
resistance of C. chalcites to several pesticides has been
recorded, e.g. in Israel (Broza and Sneh 1994). Several
publications deal with natural enemies such as Mete-
orus gyrator (Thunberg) (Hymenoptera: Braconidae)
(Bell et al. 2000), Cotesia marginiventris (Cresson)
(Hymenoptera: Braconidae) (Messelink 2002; Lobo
Lima and van Harten 1985), Podisus maculiventris
(Say) and P. nigrispinus (Dallas) (Heteroptera: Pent-
atomidae) (Vacante et al. 1996; De Clercq et al. 1998;
Bolckmans and Tetteroo 2002) but biological control is
not yet commercially available (Cabello 2009; del Pino
et al. 2011).
Foreign exploration was therefore instigated in
2008 for parasitoids suitable for the control of C. chal-
cites and T. absoluta, for use initially in Spain, but
which also would be appropriate for application
elsewhere in the increasing range of T. absoluta
in particular. The Canary Islands archipelago was
selected as an appropriate area for exploration, having
a wide range of diverse ecosystems, many approxi-
mating to both the original and invasive ranges of
T. absoluta.
Materials and methods
Study site
The Canary Islands are a volcanic archipelago which
includes seven islands and four islets. Although
politically part of Spain, they are situated in the
eastern subtropical North Atlantic (27°370–29°270N,
13°200–18°200W) near the Western Sahara region of
West Africa. The Canarian archipelago experiences a
Mediterranean-type climate of hot, dry summers and
wet, warm winters. It is, however, characterised by a
pronounced ecological heterogeneity. Whereas the
easternmost islands of the Canaries, Fuerteventura
and Lanzarote, exhibit a desert-like climate and
vegetation, in the western islands of Gran Canaria,
Tenerife, El Hierro, La Gomera and La Palma the
altitudinal zonation of vegetation is particularly
marked, and strongly differentiated between the
22 A. Polaszek et al.
123

windward and leeward sides of the islands (Ferna
´n-
dez-Palacios and Nicholas 1995).
Field collection of Trichogramma
Parasitoids were obtained from Lepidoptera eggs
collected on leaves of banana (Musa acuminata
Colla), tomato (Solanum lycopersicum L.), squash
(Cucurbita maxima Duch. ex Lam.), sweet pepper
(Capsicum annuum L.), green beans (Phaseolus
vulgaris L.) and some associated weeds (Nicotiana
glauca Graham, Solanum nigrum L.). Any emergent
parasitoids were either collected directly into 96%
ethanol in 1.5 ml Eppendorfs, or allowed to mate
and exposed to fresh Ephestia kuehniella Zeller
eggs in the laboratory in order to initiate a culture.
After possible mating and oviposition, these indi-
viduals were also collected into 96% ethanol.
Preserved specimens were carefully labelled with
details of the original collection locality, host egg
species and host plant, and date of collection.
Specimens were then sent to the first author (AP)
for identification.
Collection localities
Specimens of Trichogramma species were success-
fully obtained from the localities in Table 1.
Morphological identification
Dried specimens were initially mounted on card
squares and then on microscope slides using the
method described by Noyes (1982) with some
modifications as follows: for maceration, specimens
were heated in 10% KOH at 80°C for 10 min. This
was done without prior dissection, except for removal
of the wings. Following neutralisation with glacial
acetic acid, specimens were washed in distilled water
for 1 h, then dehydrated for 5 min in graded alcohols
of the following concentrations: 35, 70, 85, 100%.
After clearing in clove oil and allowing for evapo-
ration of alcohol, specimens were dissected in
Canada Balsam. The antennae, head, genitalia (males
only) and remaining body parts were mounted
separately on a single slide, together with the
previously mounted wings. For male mounts, the
aedeagus was removed from the genital capsule.
Identification of Trichogramma species relies very
heavily on examination of the male genitalia and to a
lesser extent the male antennae. For this reason males
were preferentially selected for this complex and
time-consuming procedure. Where possible, females
corresponding to species suggested by male identifi-
cation were later selected and mounted. Species were
initially identified based on morphology, using a
combination of the following published taxonomic
accounts: Lin (1994), Nagaraja and Nagarkatti (1970),
Nagarkatti and Nagaraja (1971), Pinto (1999), Pintu-
reau and Babault (1988) and Pintureau (2008). The
collections of the Natural History Museum (London)
contain extensive reference material of Trichogram-
ma species, including four of the five species in this
study, as well as paratypes of two of them. All
available relevant material was examined.
For preparation of the figures below, a fore wing,
the antennae and genitalia were scanned with a Leitz
Dialux compound microscope using Nomarski Dif-
ferential Interference contrast illumination, enabling
separate scanning of the upper and lower parts of
the antennae and genitalia. Scanned sections were
stacked and combined using AutoMontage
Ò
soft-
ware, and the final images edited with Adobe
Photoshop CS4
Ò
.
All specimens from the morphological study are
deposited in the Natural History Museum, London,
UK (NHM). Three voucher specimens of the T. achae-
ae population used subsequently for biocontrol of
Table 1 Collection localities of Trichogramma species in the
Canary Islands
Island Locality Coordinates
Gran Canaria Arucas 28°0705800N15°3005400W
El Hierro Frontera 27°4605100N18°1301100W
La Palma El Remo 28°3302100N17°5301700W
Fuencaliente 28°2804300N17°5104200W
Bren
˜a Baja 28°3604400N17°4503100W
Los Llanos 28°3702700N17°5505300W
Tenerife Caldera del Rey 28°0403100N16°4301500W
Cueva del Polvo 28°1304400N16°4905700W
Guargacho 28°0105200N16°3903200W
Guı
´a de Isora 28°1102200N16°4803400W
Hoya Meleque 28°2402200N16°3303100W
Pajalillos 28°3104200N16°2301300W
Valle de Guerra 28°3202600N16°2300700W
Molecular and morphological diagnoses of five species of Trichogramma 23
123

T. absoluta in Spain also had DNA extracted using a
non-destructive protocol which is also preserved (at
-80°C) in the Molecular Collections Facility (MCF)
at NHM (Figs. 1,2).
Morphological terminology follows Pinto (1999).
The specialised terminology of the male genitalia
and antenna is given in Figs. 3–8 (modified from
Pinto 1999). Host records are based largely on
those in the Universal Chalcidoidea Database (http://
www.nhm.ac.uk/research-curation/research/projects/
chalcidoids), modified by Polaszek (2010) and the
present study.
Molecular identification
39 samples were selected from the original samples
and sent to the 2nd and 3rd authors PFR-J and RS
(UC Riverside, USA) for DNA extraction and
amplification of the ITS2 region. DNA was extracted
from single female wasps using the Chelex method.
The wasps were crushed in a 0.5 ml micro-centrifuge
tube with a closed Pasteur pipette and ground in 60 ll
5% Chelex-100 and 2 ll of Proteinase K (20 mg
ml
-1
), followed by incubation for 1 h at 55°C, and
finally for 10 min at 99°C.
The polymerase chain reaction (PCR) was used to
amplify the ITS2 region of each specimen using
primers described in Stouthamer et al. (1999). Reac-
tions were performed in 25 ll volumes containing: 19
Thermopol Reaction Buffer (New England BioLabs,
Ipswich, MA, USA); 200 lM each of dATP, dCTP,
and dGTP; 400 lM dUTP; 0.2 lM each primer; 1 U
Taq polymerase (NEB); 2 ll template DNA (concen-
tration not determined). Amplification was achieved
using a Mastercycler ep gradient S (Eppendorf North
America Inc., New York, NY, USA), programmed for:
3 min at 95°C; followed by 37 cycles of 45 s at 94°C,
45 s at 53°C, and 1 min at 72°C; and a final step of
3 min at 72°C. PCR products were visualized after
electrophoresis on 1.5% agarose gel stained with
ethidium bromide, to confirm amplification.
Species were identified based on the size of their
ITS2 PCR product (e.g. Kumar et al. 2009; Silva
et al. 1999; Sumer et al. 2009; Thomson et al. 2003).
The PCR products of a subset of 14 samples were
directly sequenced in both directions at the Institute
for Integrative Genome Biology, University of
California, Riverside, USA. Sequences were aligned
manually using BioEdit version 7.0.9.0 (Hall 1999),
Fig. 1 First 97 bases in the aligned ITS2 fragment for six Trichogramma species
24 A. Polaszek et al.
123

and compared to the ITS2 sequences of known
Trichogramma spp. (Figs. 1,2). Positions 1–82 in the
manually aligned sequence are identical to the result
obtained by aligning in BioEdit with the option
‘‘accessory application—ClustalW multiple align-
ment’’ using default parameters. Complete ITS2
sequences of 16 specimens, including all the species
treated here, have been deposited in Genbank under
accession numbers JF415934–JF415939.
Results
Morphological examination of antennae, male geni-
talia and wings enabled three morphospecies to be
easily distinguished: T. achaeae,T. bourarachae and
T. euproctidis. All three are new records for the
Canary Islands, from where no Trichogramma
species had been recorded previously. T achaeae,
described originally from India, proved in laboratory
experiments to be a very promising candidate for the
inundative biological control of T. absoluta (Cabello
et al. 2009a) and subsequently also in field experi-
ments (Vila et al. 2010). T. evanescens and T. near
brassicae could also be distinguished to some
extent morphologically, but with some difficulty.
PCR confirmed the presence of the three morpholog-
ically distinct species, and indicated the presence of
the two others: T. evanescens and two specimens of
an unknown species near T. brassicae.
In just two collections out of the 39 examined was
there disagreement between the results of the molec-
ular and morphological identification. One of these
concerned T. bourarachae (evident from morphol-
ogy) identified with PCR as T. euproctidis, and the
second a collection of T. achaeae misidentified as
T. evanescens. It seems most probable that these two
collections originally contained a mixture of species,
possibly due to contamination. Overall, the congru-
ence between morphological and molecular (PCR)
identification (95%) was extremely high.
All five species can be distinguished by their ITS2
sequences (Figs. 1,2) but also based on the size of the
amplified PCR product: T. euprocidis: 377–378 nt;
T. evanescens; 430–432 nt; T. near brassicae; 411 nt;
T. achaeae; 518–519 nt; T. bourarachae:*560 nt. The
sequence of T. evanescens from the Canary Islands
differs very slightly from ‘‘standard’’ T. evanescens,
represented in Figs. 1and 2by the specimen from
Oman. T. near brassicae differs by approximately 7%
in ITS2 sequence from the standard T. brassicae,
represented in Figs. 1and 2by the specimen from
Turkey.
Discussion
Trichogramma in the Canary Islands
It is perhaps surprising that after just a few months of
field collecting, five Trichogramma species have been
discovered in the Canary Islands where no Trichogram-
ma species had been recorded previously. Furthermore,
the species have particular characteristics regarding
their known previous distribution. T. achaeae was
described from Bangalore in Southern India (Nagaraja
and Nagarkatti 1970), but has since been recorded from
many localities around the world, including (recorded
here for the first time) Hawaii. It appears, therefore,to be
a good coloniser, being highly adaptive. This ability
raises legitimate concerns about its possible non-target
effects in areas of its spread, either accidental or
deliberately (Desneux et al. 2010). Of the possible
non-target Lepidoptera genera listed by these authors,
Fig. 2 Bases 150–180 in the aligned ITS2 fragment for four Trichogramma species
Molecular and morphological diagnoses of five species of Trichogramma 25
123

Figs. 3–8 Morphological
terminology for antennal
(Fig. 3) and male genitalia
(Figs. 4–8) characters
(after Pinto 1999)
26 A. Polaszek et al.
123

none has so far been recorded as a host of T. achaeae (see
below), but host range testing will be necessary prior to
future field releases.
T. bourarachae was described from Morocco, and
is now known additionally from Egypt, Tunisia,
Portugal (Polaszek 2010) and the Canary Islands.
T. euproctidis and T. evanescens are very widespread
Old World species, although many published records
are certainly based on misidentifications. Finally,
T. near brassicae appears to be a new species,
possibly endemic to the Canary Islands, but requiring
further study.
T. achaeae as a biocontrol agent of Chrysodeixis
chalcites and Tuta absoluta
The frequency of T. achaeae attacking eggs of
C. chalcites in the field suggests good potential for
this species as a control agent in banana crops, and
field trials against this host are ongoing (del Pino,
unpublished data). However, against T. absoluta,
T. achaeae is already proving an effective biocontrol
agent (Cabello et al. 2009a). Desneux et al. (2010)
found that the release of T. achaeae at a rate of
50 adults m
-2
two times a week during the first
ten weeks of the tomato growing season significantly
reduced the number of T. absoluta larvae, leaf mines
and damaged fruits, compared with control plots. Vila
et al. (2010) found that releases of 25 T. achaeae
adults m
-2
per week during seven weeks also
provides significant control of T. absoluta. These
application rates appear to be high, but are within
what is commonly used in inundative biological
control programs in other crops (Mills 2010). For use
in the field T. achaeae is mass-reared on eggs of
Ephestia kuhniella, and parasitized eggs are distrib-
uted to growers in user-friendly dispensers (Desneux
et al. 2010) (Figs. 9–13,14–18).
Species accounts
Trichogramma achaeae Nagaraja & Nagarkatti
1970 (Figs. 9,14,19)
Synonymy
Trichogramma achaea Nagaraja & Nagarkatti
Diagnosis. Flagellum of male antenna (Fig. 9)
2.1 ±0.19length of scape (n=9). Flagellar length/
flagellar width (not including setae) 5.9 ±0.6 (n=9);
max. flagellar setal length/basal flagellar width
2.7 ±0.4 (n=9). Terminal placoid sensilla extend-
ing beyond the end of the flagellum. Genital capsule
length (Fig. 14a) 2.8 ±0.39width (n=9), sides very
slightly narrowed at level of IVP. AD/GL =0.2
(n=9); AW/GW =0.5 ±0.2 (n=9); DAL/GL =
0.5 (n=9). IVP (Fig. 10a) very small. DLA (Fig. 14b)
originating at middle of GC, linguiform and parallel-
sided for some of its length; rounded at apex. Shoulders
not present at base of DLA. Aedeagus (Fig. 14c) length
equal to GL; 1.9 ±0.39apodemes (n=7).
Distribution. Asia: China, India, New Caledonia,
Russia; Europe: France (introduced), Portugal
(Ac¸ores), Russia, Spain (introduced to mainland.
Canary Islands: El Hierro, Gran Canaraia, La Palma,
Tenerife); Africa: Cape Verde; Americas: Argentina,
Barbados, Chile, Trinidad & Tobago, Hawaii, USA.
Hosts. Diptera: Anthomyiidae: Atherigona soccata;
Lepidoptera: Gelechiidae: Pectinophora gossypiella;
Sitotroga cerealella,Tuta absoluta; Geometridae: Boar-
mia variegata;Noctuidae:Achaea janata;Anticarsia
gemmatalis;Chrysodeixis chalcites;Cornitoplusia sp.;
Earias sp.; E. insulana;E. vittella;Helicoverpa armi-
gera;H. zea;Mamestra brassicae;Spodoptera sp.;
Tiracola plagiata;Trichoplusia ni; Notodontidae: Clos-
tera cupreata; Oecophoridae: Opisina arenosella;
Pieridae: Eurema sp. Pyralidae (including Crambridae):
Chilo partellus;Corcyra cephalonica;Loxostege sticti-
calis; Sphingidae: Acherontia styx;Agrius convolvuli;
Tortricidae: Cydia koenigana, Laspeyresia koenig-
ana; Yponomeutidae: Plutella xylostella.Nymphalidae:
Danaus plexippus; Lyceanidae: Lampides boeticus:
Material examined (specimens marked with ‘‘*’’
were confirmed by sequencing ITS2): Paratypes 9 #:
India: West Bengal, Kalimpong iv.1965 (V.R. Pha-
lak) ex egg Achaea janata CIE 2801 (NHM). 1#*
Spain: Islas Canarias, Tenerife, Valle de Guerra,
26/01/2009 UTM 28R 0364467 3157871 Solanum
lycopersicum F05/09-55 ex C. chalcites;1$1#
Tenerife, Guargacho 23/10/2008 GU05-06 Musa
acuminata ex C. chalcites;1#* Tenerife, Gu
¨ı
´ade
Isora, 10/03/2009 UTM 28R 0322368 3119543 TF18/
09-29 Nicotiana glauca ex Cornitoplusia sp.; 1#*La
Palma, Fuencaliente, 18/05/2009 UTM 28R 0219803
3153587 M. acuminata LP10/09-07 ex C. chalcites;
1#* same data but 20/03/2009 LP04/09-05; 1#
same data but S. lycopersicum LP11/09-96; 1#*La
Palma, Fuencaliente, 24/04/2009 UTM 28R 0219803
Molecular and morphological diagnoses of five species of Trichogramma 27
123

Figs. 9–13 Trichogramma male antennae, inner (a) and outer (b) aspects. Figure 9T. achaeae; Fig. 10 T. bourarachae;
Fig. 11 T. euproctidis; Fig. 12 T. evanescens; Fig. 13 T. near brassicae
28 A. Polaszek et al.
123

3153587 S. lycopersicum LP10/09-07 ex C. chalcites;
1#same data but LP09/09-29; 1#same data but
25/07/2008 LP17-21; 1#* same data but 04/06/2009
LP16/09-37; 2#* same data but Phaseolus vulgaris
LP17/09-44; 1#same data but LP17/09-61; 2#*La
Palma, Los Llanos, 14/07/2009 N. glauca LP21/09-
32 ex C. chalcites;2#* La Palma, El Remo, 18/05/
2009 UTM 28R 0217434 3162190 Capsicum annuum
LP12/09-04 ex C. chalcites;1#* same data but
20/03/2009 LP02/09-01; 1#same data but LP13/09-
Figs. 14–18 Trichogramma species, male genitalia. aParameres, volsellae, intervolsellar process; bdorsal lamina; c: aedeagus.
Figure 14 T. achaeae; Fig. 15 T. bourarachae; Fig. 16 T. euproctidis; Fig. 17 T. evanescens; Fig. 18 T. near brassicae
Molecular and morphological diagnoses of five species of Trichogramma 29
123

13; 1$1#same data but 30/09/2008 LP22-21; 1#
same data but 30/09/2008 LP22-01 M. acuminata ex
C. chalcites;2#* La Palma, Bren
˜a Alta, 24/04/2009
28R UTM 0230252 3168161 M. acuminata LP08/09-
12 ex C. chalcites;1#same data but 05/08/2009
LP23/09-51; 1$*1#Gran Canaria, Arucas, 11/05/
2009 UTM 28R 0449402 3112042 S. lycopersicum
GC09/09-16 ex C. chalcites;1$3#* El Hierro,
Frontera, 14/12/2008 M. acuminata, HR10/08-09 ex
C. chalcites;2#* same data but 22/01/2009 HR01/
09-07, HR01/09-10.
Trichogramma bourarachae Pintureau & Babault
1988 (Figs. 10,15,20)
Diagnosis. Flagellum of male antenna (Fig. 10)2.09
length of scape (n=4). Flagellar length/flagellar width
(not including setae) 5.9 ±0.6 (n=4); max. flagellar
setal length/basal flagellar width 2.3 ±0.2 (n=3).
Terminal placoid sensilla extending beyond the end
of the flagellum. Genital capsule length (Fig. 15a)
3.7 ±0.39the width (n=4), sides strongly narrowed
at level of IVP. AD/GL =0.2 (n=4); AW/GW =0.6
(n=4); DAL/GL =0.5 (n=4). IVP (Fig. 15a) small.
DLA (Fig. 15b) originating in basal third of GC, narrow
and pointed at apex. Shoulders not present at base
of DLA. Aedeagus (Fig. 15c) length equal to GL;
2.4 ±0.29apodemes (n=4).
Distribution. Europe: Portugal; Spain (including
Canary Islands, Tenerife). Africa: Egypt, Morocco,
Tunisia.
Hosts. Lepidoptera: Lymantriidae: Euproctis
chrysorrhoea;Noctuidae:Chrysodeixis chalcites;
Helicoverpa armigera; Nymphalidae: Vanessa cardui;
Figs. 19–23 Trichogramma species, fore wing. Figure 19 T. achaeae; Fig. 20 T. bourarachae; Fig. 21 T. euproctidis; Fig. 22
T. evanescens; Fig. 23 T. near brassicae
30 A. Polaszek et al.
123

Pyralidae: Ectomyelois ceratoniae;Ephestia kuehni-
ella;Palpita unionalis; Yponomeutidae: Prays oleae;
Neuroptera: Chrysopidae: Chrysoperla carnea.
Material examined (specimens marked with ‘‘*’’
were confirmed by sequencing ITS2): Paratype 1#:
Marocco: Baume (B.Pintureau, M. Babault) ex egg
E. kuehniella (NHM). 1#* Spain: Islas Canarias,
Tenerife, Valle de Guerra, 28/07/2009 UTM 28R
0364195 3154856N. gl auca TF43/09-25 ex C. chalcites;
1$Tenerife, Cueva del Polvo 16/03/2009 UTM
28R 0320185 3123941 N. glauca TF20/09-08 ex
C. chalcites;samedatabut:1#N. glauca TF20/09-19
ex Cornitoplusia sp.; 2$*1#*N. glauca TF20/09-33 ex
C. chalcites.
Trichogramma euproctidis (Girault, 1911)
(Figs. 11,16,21)
Synonymy
Pentarthron euproctidis Girault
Trichogramma turkestanica Meyer
Trichogramma meyeri Sorokina
Diagnosis. Flagellum of male antenna (Fig. 11)
1.9 ±0.29length of scape (n=10). Flagellar length/
flagellar width (not including setae) 5.9 ±0.6
(n=10); max. flagellar setal length/basal flagellar
width 3.3 ±0.4 (n=10). Terminal placoid sen-
silla extending beyond the end of the flagellum.
Genital capsule length (Fig. 16) 2.7 ±0.39the width
(n=10), sides very slightly narrowed at level of IVP.
AD/GL =0.2 (n=9); AW/GW =0.6 (n=9);
DAL/GL =0.6 (n=9). IVP (Fig. 16a) large, prom-
inent. DLA (Fig. 16b) originating at middle of GC,
triangular and rounded at apex. Shoulders present at
base of DLA. Aedeagus (Fig. 16c) length equal to GL;
2.2 ±0.19apodemes (n=10).
Distribution. Asia: Armenia, China, Japan,
Kazakhstan, Russia, Tadzhikistan, Turkmenistan,
Turkey, Uzbekistan, Vietnam; Europe: Belarus,
Bulgaria, Denmark, France, Greece, Italy, Moldova,
Portugal, Russia, Spain (incl. Canary Islands: Gran
Canaria, Tenerife), Ukraine; Africa: Egypt, Morocco;
Americas: Argentina, Chile, Cuba, Peru.
Hosts. Diptera: Anthomyiidae: Erioischia brassi-
cae; Lepidoptera: Gelechiidae: Sitotroga cerealella;
Lymantriidae: Euproctis chrysorrhoea;Nygmia
phaeorrhoea;Orgyia antiqua; Noctuidae: Agrotis
segetum;Amathes c-nigrum;Chrysodeixis chalcites;
Helicoverpa armigera;Mamestra brassicae;Sesamia
nonagrioides;Syngrapha circumflexa; Nymphalidae:
Vaness cardui; Pieridae: Pieris brassicae; Pyralidae
(incl. Crambidae): Chilo agamenon;Corcyra cepha-
lonica;Ephestia kuehniella;Loxostege sticticalis;
Margaronia quadristigmalis;Mescinia peruella;
Ostrinia nubilale; Sphingidae: Agrius convolvuli;
Tortricidae: Archips rosanus;Cydia pomonella;
Epichoristodes acerbella;Grapholitha molesta;
Lobesia botrana; Yponomeutidae: Plutella maculi-
pennis;Prays oleae. Neuroptera: Chrysopidae:
Chrysoperla carnea.
Material examined (specimens marked with ‘‘*’’
were confirmed by sequencing ITS2): 1#* Spain:
Islas Canarias, Tenerife, Hoya Meleque, 04/02/2009
UTM 28R 0347304 3143191 Solanum lycopersicum
TF08/09-41 ex Chrysodeixis chalcites; same data but:
1#* 31/03/2009 S. lycopersicum TF26/09-27 ex C.
chalcites;1#* Tenerife, Guargacho 13/02/2009 UTM
28R 0336906 3101763 Musa acuminata TF10/09-11
ex C. chalcites; same data but: 1#23/10/2008 Musa
acuminata Gu05-16 ex C. chalcites;1#Tenerife,
Gu
¨ı
´a de Isora 10/03/2009 TF18/09-47 UTM 28R
0322368 3119543 N. glauca ex Cornitoplusia sp.;
same data but: 1#N. glauca TF18/09-38 ex Corni-
toplusia sp.; 2#*1$* Tenerife, Caldera del Rey
25/06/2009 TF41/09-77 UTM 28R 0330902 3106753
S. lycopersicum ex C. chalcites; same data but:1#*
17/04/2009 S. lycopersicum TF30/09-43 ex C. chal-
cites;1$Tenerife, Cueva del Polvo 16/03/2009 UTM
28R 0320185 3123941 TF20/09-33 N. glauca ex
Cornitoplusia sp.
Trichogramma evanescens Westwood, 1833
(Figs. 12,17,22)
Synonymy
Calleptiles latipennis Haliday
Calleptiles vitripennis (Walker)
Pentarthron carpocapsae Schreiner
Pentarthron carpocapsai Schreiner
Pteroptrix evanescens (Westwood)
Trichogramma barathrae Skriptshinsky
Trichogramma cacoeciae pini Meyer
Trichogramma carpocapsae (Schreiner)
Trichogramma evanescens piniperda Wolff
Trichogramma pini Meyer
Molecular and morphological diagnoses of five species of Trichogramma 31
123

Trichogramma piniperdae Wolff (Pintureau (2008)
treats T. piniperda as a valid species)
Trichogramma rhenana Voegele
´& Russo
Trichogramma rhenanum Voegele
´& Russo
Trichogramma vitripennis Walker
Trichogramma vitripenne Walker
The true identity of T. evanescens is still uncertain
at the time of writing, but for practical reasons, most
workers follow Pintureau’s interpretation of this
highly cited species (see Pintureau 2008).
Diagnosis. Flagellum of male antenna (Fig. 12)
2.1 ±0.19length of scape (n=3). Flagellar length/
flagellar width (not including setae) 5.6 ±0.1 (n=4);
max. flagellar setal length/basal flagellar width
3.1 ±0.2 (n=4). Terminal placoid sensilla extend-
ing beyond the end of the flagellum. Genital capsule
length (Fig. 17) 2.8 ±0.29the width (n=4), sides
very slightly narrowed at level of IVP. AD/GL =0.3
(n=4); AW/GW =0.6 (n=4); DAL/GL =0.6
(n=4). IVP (Fig. 17a) large, prominent. DLA
(Fig. 17b) originating at middle of GC, triangular and
rounded at apex. Shoulders present at base of DLA.
Aedeagus (Fig. 17c) length equal to GL; 2.2 ±0.19
apodemes (n=4).
Distribution. Asia: Armenia, Azerbaijan, China,
Georgia, India, Iran, Israel, Kazakhstan, Oman,
Pakistan, Philippines, Russia, Sri Lanka, Turkey,
Turkmenistan, Uzbekistan, Vietnam; Europe: Aus-
tria, Belarus, Belgium, Bulgaria, Czech Republic,
Denmark, France, Germany, Hungary, Ireland, Italy,
Lithuania, Macedonia, Moldova, Netherlands,
Poland, Portugal (including Madeira), Romania,
Russia, Serbia, Slovakia, Spain (incl. Canary islands:
La Palma, Tenerife), Sweden, Switzerland, Turkey,
Ukraine, United Kingdom, Yugoslavia (former, pre
1991). Africa: Comores, Egypt, Libya, Madagascar,
Mauritius, Morocco.
Hosts. Coleoptera: Bruchidae: Bruchus obtectus;
Chrysomelidae: Cassida deflorata;C. nebulosa;C. no-
bilis;C. vittata;Donacia simplex; Curculionidae:
Rhynchaenus testaceus;Dermestidae: Dermestes mac-
ulatus; Rhynchitidae: Rhynchites auratus;R. betulae;
Tenebrionidae: Tribolium castaneum; Diptera:
Anthomyiidae: Atherigona soccata;Pegomya betae;
P. hyoscyami; Stratiomyiidae: Oxycera sp.; Stratiomys
sp.; Syrphidae: Melanostoma mellinum;Paragus
quadrifasciatus;Syrphus sp.; S. balteatus;S. pyrastri;
S. vitripennis; Tabanidae: Chrysops sp.; C. caecutiens;
C. relictus;Tabanus sp.; Hemiptera: Cimicidae: Cimex
lectularius; Hymenoptera: Pamphiliidae: Acantholyda
erythrocephala;A. posticalis;A. stellata;Cephalcia
abietis;C. arvensis;C. signata; Tenthredinidae: Cal-
iroa cerasi;Croesus septentrionalis;Emphytus tener;
Pteronidea ferruginea;P. ribesii; Lepidoptera: Arctii-
dae: Arctia caja;Diacrisia obliqua;Eilema sp.;
Hyphantria cunea;Spilosoma sp.; Bombycidae:
Rondotia menciana; Danaidae: Danaus chrysippus;
Gelechiidae: Pectinophora sp.; P. gossypiella;Phthor-
imaea operculella;Sitotroga cerealella;Geometridae:
Boarmia grisescens;Bupalus piniarius;Cidaria
bilineata;C. didymata;Crocallis elinguaria;Erannis
defoliaria;Lambdina fiscellaria;Operophtera
brumata; Glyphipterygidae: Anthophila atrilineata;
Lasiocampidae: Cosmotriche potatoria;Dendrolimus
pini;D. punctatus;D. segregatus;D. spectabilis;
Malacosoma disstria;M. neustria; Leptidae: Atherix
sp.; Lycaenidae: Cacyreus marshalli;Thecla betulae;
Virachola livia; Lymantriidae: Arctornis chrysor-
rhoea;Euproctis lunata;E. phaeorrhoea;Laelia
salicis;Lymantria dispar;L. monacha;Nygmia phae-
orrhoea;Orgyia antiqua;O. gonostigma;Stilpnotia
salicis; Noctuidae: Acontia luctuosa;Acronicta aceris;
A. major;A. rumicis;A. tridens;Agrotis exclamationis;
A. ipsilon;A. segetum;Amathes c-nigrum;Antitype
flavicincta;Apopestes spectrum;Autographa sp.;
A. gamma;Barathra brassicae;Catocala elocata;
Chrysodeixis chalcites;Cirrhia gilvago;Discestra
trifolii;Earias sp.; E. cupreoviridis;E. fabia;E. insul-
ana;Euxoa obelisca;E. segetum;Gonospileia glyph-
ica;Helicoverpa armigera;H. assulta;H. virescens;
H. zea;Mamestra sp.; M. brassicae;M. oleracea;
M. trifolii;Naranga aenescens;Noctua pronuba;Oria
musculosa;Panolis flammea;Parallelia algira;Phala-
ena typica;Phlogophora meticulosa;Phytometra
gamma;Plusia gamma;Polia oleracea;P. pisi;
P. suasa;Pyrrhia umbra;Rivula atimeta;Sarrothripus
musculana;Scotia ipsilon;Sesamia cretica;S. non-
agrioides;Spaelotis pronubana;Spodoptera sp.;
S. littoralis;S. litura;Tholera popularis;Trachea
triplicis;Trichoplusia ni;Triphaena pronuba;Noto-
dontidae:Lampronadata cristata;Phalera bucephala;
P. bucephaloides;Thaumetopoea pityocampa; Nymp-
halidae: Nymphalis polychloros; Oecophoridae:
Depressaria nervosa;Endrosis lactella;Opisina
arenosella; Papilionidae: Iphiclides podalirius;Papi-
lio polytes; Pieridae: Aporia crataegi;Leptidea sina-
pis;Pieris sp.; P. brassicae;P. daplidice;P. napi;
32 A. Polaszek et al.
123

P. rapae; Pyralidae: (incl. Crambidae): Achroia
grisella;Anagasta sp.; Cadra cautella;Chilo sp.;
C. agamemnon;C. indicus;C. infuscatellus;C. partel-
lus;C. sacchariphagus;C. simplex;C. suppressalis;
Cnaphalocrocis medinalis;C. cephalonica;Crambus
geniculeus;Diatraea sp.; Ectomyelois ceratoniae;
Ephestia sp.; E. calidella;E. cautella;E. elutella;
E. kuehniella;Etiella zinckenella;Evergestis forfical-
is;Galleria sp.; G. mellonella;Glyphodes pyloalis;
Hymenia recurvalis;Loxostege sticticalis;Marasmia
patnalis;Maruca vitrata;Ostrinia furnacalis;O. nu-
bilale;Palpita unionalis;Phlyctaenia forficalis;
Plodia interpunctella;Pyrausta machaeralis;Sale-
bria semirubella;Spectrobates ceratoniae; Saturnii-
dae: Attacus cynthia;A. ricini; Sphingidae: Celerio
lineata;Manduca sexta;Smerinthus populi;Sphinx
pinastri; Tortricidae: Adoxophyes orana;A. reticul-
ana;Archips crataeganus;A. pronubana;A. rosanus;
Cacoecia rosanus;Cacoecimorpha pronubana;
Carpocapsa pomonella;Choristoneura fumiferana;
Clysia ambiguella;Cnephasia longana;Cneph-
asia pumicana;Cydia funebrana;C. pomonella;
Epichoristodes acerbella;Epinotia pygmaeana;
E. tedella;Eupoecilia ambiguella;Grapholitha de-
lineana;Grapholitha funebrana;G. molesta;Gypso-
noma aceriana;Homona coffearia;Laspeyresia
microgrammana;L. molesta;L. nigricana;Lobesia
botrana;Pandemis chondrillana;Pandemis hepar-
ana;Petrova resinella;Rhopobota naevana;Rhya-
cionia buoliana;Sparganothis pilleriana;Tetramoera
schistaceana;Zeiraphera diniana; Yponomeutidae:
Acrolepiopsis assectella;Argyresthia conjugella;
Plutella sp.; P. maculipennis;P. xylostella;P. citri;
Prays oleae; Zygaenidae: Theresimima ampelophaga;
Zygaena sp.; Megaloptera: Sialidae: Sialis lutaria;
Neuroptera: Chrysopidae: Chrysoperla carnea;
C. ventralis;Chrysoperla sp.; Nothochrysa italica.
Material examined (specimens marked with ‘‘*’’
were confirmed by sequencing ITS2): 1#* Spain:
Islas Canarias, La Palma, El Remo, 18/05/2009 28R
0217434 UTM 3162190 Cucurbita maxima LP13/
09-30 ex Chrysodeixis chalcites; same data but:
1#* 20/03/2009 Nicotiana glauca LP03/09-12 ex
C. chalcites;1#* Tenerife, Pajalillos 18/02/2009 28R
0364277 UTM 3156545 Solanum lycopersicum
LP12/09-01 ex C. chalcites.
Trichogramma near brassicae (Figs. 13,18,23)
Diagnosis. Flagellum of male antenna (Fig. 13) 1.99
length of scape (n=1). Flagellar length/flagellar
width (not including setae) 4.7; max. flagellar setal
length/basal flagellar width 3.6 (n=1). Terminal
placoid sensilla extending beyond the end of the
flagellum. Genital capsule length (Fig. 18a) 3.19the
width (n=1), sides narrowed at level of IVP. AW/
GW =0.5 (n=1); DAL/GL =0.5 (n=1). IVP
(Fig. 18a, b) large, prominent. DLA (Fig. 10b) orig-
inating at middle of GC, triangular and rounded at
apex. Shoulders present at base of DLA. Aedeagus
(Fig. 18c).
Distribution. Europe: Spain (Canary Islands: Gran
Canaria).
Host. Lepidoptera: Noctuidae: Chrysodeixis
chalcites.
Material Examined (confirmed by sequencing
ITS2): 2#* Spain: Gran Canaria, Arucas, 11/05/
2009 UTM 28R 0449402 3112042 Musa acuminata
GC07/09-17 ex Chrysodeixis chalcites.
Comments. The authors are currently undertaking
further examination of this species, including cross-
breeding experiments. The species will be formally
described in the near future.
Acknowledgments We are grateful to AgroBio S.L. and
ASPROCAN for their financial support and collaboration
during Trichogramma field collection. The research of M. del
Pino was financially supported by a pre-doctoral fellowship
granted by ‘‘Instituto Nacional de Investigacio
´n y Tecnologı
´a
Agraria y Alimentaria (INIA)’’. The paper benefited greatly
from comments by two anonymous reviewers
References
Alonso A (2009) Eficacia del control quı
´mico y ubicacio
´nde
trampas para monitoreo de Chrysodeixis chalcites (Esper,
1789) en cultivos de platanera de Canarias. Trabajo Fin de
Carrera. Escuela Te
´cnica Superior de Ingenierı
´a Agraria.
Universidad de La Laguna, La Laguna
Bell HA, Smethurst F, Marris GC, Edwards JP (2000) Mete-
orus gyrator: a potential biocontrol agent against glass-
house noctuid pests? In: The BCPC conference: pests and
diseases, vol 1. Proceedings of an international conference
held at the Brighton Hilton Metropole Hotel, Brighton,
UK, 13–16 November 2000, pp 291–296
Molecular and morphological diagnoses of five species of Trichogramma 33
123

Bolckmans KJF, Teteeroo ANM (2002) Biological pest control
in eggplants in the Netherlands. IOBC/WPRS Bull 25(1):
25–28
Branco MC, Franca FH (1993) Susceptibility of three popu-
lations of Scrobipalpuloides absoluta (Lep.: Gelechiidae)
to cartap. Hortic Bras 11:32–34
Broza M, Sneh B (1994) Bacillus thuringiensis spp. kurstaki as
an effective control agent of lepidopteran pests in tomato
fields in Israel. J Econ Entomol 87(4):923–928
Bueno VHP (2005) Implementation of biological control in
greenhouses in Latin America: how far are we? In: 2nd
International symposium on Biological control of arthro-
pods. USDA Forest Service Publication FHTET-2005-08,
pp 531–537
Cabello T (1986) Plagas de lepido
´pteros en cultivos del Valle
del Guadalquivir. Actas de las VIll Jornadas de la Aso-
ciacio
´n Espan
˜ola de Entomologı
´a, Sevilla, pp 839–848
Cabello T (1988) Especies de noctuidos (Lep.: Noctuidae) de
intere
´s agrı
´cola en la Vega de Granada y su fenologı
´a.
Actas del III Congreso Ibe
´rico de Entomologı
´a, Granada,
pp 925–936
Cabello T (2009) Control biolo
´gico de Noctuidos y otros
Lepido
´pteros. In: Jacas J, Urbaneja A (eds) Control biol-
o
´gico de Plagas. Phytoma, Valencia, Spain, pp 279–306
Cabello T, Gonza
´lez MP, Justicia L, Belda JE (1996) Plagas de
noctuidos (Lep.; Noctuidae) y su fenologı
´a en cultivos en
invernaderos. Informaciones Te
´cnicas 39/96. Direccio
´n
General de Investigacio
´n y Formacio
´n Agraria. Consejerı
´a
de Agricultura y Pesca. Junta de Andalucı
´a, Sevilla
Cabello T, Gallego JR, Ferna
´ndez-Maldonado FJ, Soler A,
Beltra
´n D, Parra A, Vila E (2009a) The damsel bug Nabis
pseudoferus (Hem.: Nabidae) as a new biological control
agent of the South American Tomato Pinworm, Tuta ab-
soluta (Lep.: Gelechiidae), in tomato crops of Spain.
IOBC/WPRS Bull 49:219–223
Cabello T, Gallego JR, Vila E, Soler A, Del Pino M, Carnero A,
Hernandez-Suarez E, Polaszek A (2009b) Biological con-
trol of the South American tomato pinworm Tuta absoluta
(Lep. Gelechiidae), with releases of Trichogramma
achaeae (Hym.: Trichogrammatidae) in tomato green-
houses of Spain. Integrated Control in Protected Crops,
Mediterranean Climate. IOBC/WPRS Bull 49:225–230
Cayrol RA (1972) Famille des Noctuidae. In: Balachowsky AS
(ed) Entomologie applique
´ea
`l’agriculture. Le
´pidopte
`res.
Tome II, vol 2. Masson et Cie, Parı
´s, pp 1.255–1.520
De Clercq P, Merleveden F, Mestadagh I, Vanderdurpel K,
Mahaghegh J, Degheele D (1998) Predation on the tomato
looper Chrysodeixis chalcites (Lep.: Noctuidae) by Po-
disus maculiventris and Podisus nigrispinus (Het.; Pent-
atomidae). J Appl Entomol 122:93–98
del Pino M, Carnero A, Cabello T, Herna
´ndez E (2011) La
lagarta o bicho camello, Chrysodeixis chalcites (Esper,
1789), una plaga emergente en los cultivos de platanera de
Canarias. Phytoma Esp 225:21–24
Desneux N, Wajnberg E, Kris A, Wyckhuys G, Burgio G,
Arpaia S, Narva
´ez-Vasquez CA, Gonza
´lez-Cabrera J,
Catala
´n-Ruescas D, Tabone E, Frandon J, Pizzol J, Poncet
C, Cabello T, Urbaneja A (2010) Biological invasion of
European tomato crops by Tuta absoluta: ecology, geo-
graphic expansion and prospects for biological control.
J Pest Sci 83:197–215
Domingues GR, Oliveira HN, Kasten P, Parra JRP (2003)
Control of the tomato moth Tuta absoluta (Lep.: Gele-
chiidae) by Trichogramma pretiosum released in green-
houses Egg Parasitoid News 15:26
EPPO (2005) Tuta absoluta. OEPP/EPPO Bull 35:434–435
EPPO (2008) First report of Tuta absoluta in Spain. EPPO
Reporting Service no 1 2008-01-01
Faria CA, Torres JB, Fernandez AMV, Farias AMI (2008)
Parasitism of Tuta absoluta in tomato plants by Tricho-
gramma pretiosum Riley in response to host density and
plant structures. Cienc Rural 38:1504–1509
FERA (UK Food & Environment Research Agency) (2010)
FERA confirms the first outbreak in the UK of Tuta
absoluta—the South American tomato moth. http://www.
fera.defra.gov.uk/showNews.cfm?id=402
Ferna
´ndez-Palacios JM, Nicholas JP (1995) Altitudinal pattern
of vegetation variation on Tenerife. J Veg Sci 6:183–190
Freitas FS, Torres JB, Pratissoli D, Fosse-Filho E (1994)
Control of tomato borer Srobipalpuloides absoluta in
periods of population peaks, wirth Trichogramma preti-
osum and cartap. Ceres 41:244–253
Hall TA (1999) BioEdit: a user friendly biological sequence
alignment and analysis program from Windows 95/98/NT.
Nucleic Acids Symp 41:95–98
Harakly FA, Farag SS (1975) Biological studies on the tomato
looper Chrysodeixis chalcytes (Esper) in Egypt. Bull Soc
Entomol Egypte 59:295–299
Kumar GA, Jalali SK, Venkatesan T, Stouthamer R, Niranjana
P, Lalitha Y (2009) Internal transcribed spacer-2 restric-
tion fragment length polymorphism (ITS-2-RFLP) tool
the differentiate some exotic and indigenous trichogram-
matid egg parasitoids in India. Biol Control 49:207–213
Lietti MMM, Botto E, Alzogaray RA (2005) Insecticide
resistance in Argentine populations of Tuta absoluta
(Lep.: Gelechiidae). Neotrop Entomol 34:113–119
Lin NQ (1994) Systematic studies of Chinese Trichogramm-
matidae. Contributions of the Biological Control Research
Institute, Fujian Agricultural University. Special Publi-
cation No 4. Chongqing Publishing House, Chongqing,
China (Chinese with English summary), 362 pp
Lobo Lima Ml, van Harten A (1985) Biological control of
agricultural pests on the Cape Verde Islands. Present sit-
uation and prospects for the future. Revista Investigacao
Agraria. Centro de Estudios Agrarios Ser A 1:3–10
Loginova E (1992) Some new pests of glasshouse crops in
Bulgaria and their control by an IPM programme. Bull
OEPP/EPPO Bull 22:357–361
Messelink G (2002) Biological control of caterpillars with
Cotesia marginiventris (Hym.: Braconidae) in sweet
pepper and tomato. IOBC/WPRS Bull 25(1):181–184
Mills N (2010) Egg parasitoids in biological control and inte-
grated pest management. In: Co
ˆnsoli FL, Parra JRP,
Zucchi RA (eds) Egg parasitoids in agroecosystems with
emphasis on Trichogramma. Progress in Biological Con-
trol, vol 9. Springer, Dordrecht, pp 237–266
Nagaraja H, Nagarkatti S (1970) Three new species of
Trichogramma from India. Entomophaga 14(4):393–400
Nagarkatti S, Nagaraja H (1971) Redescriptions of some
known species of Trichogramma (Hym., Trichogramm-
matidae), showing the importance of the male genitalia as
a diagnostic character. Bull Entomol Res 61:13–31
34 A. Polaszek et al.
123

Noyes JS (1982) Collecting and preserving chalcid wasps
(Hymenoptera: Chalcidoidea). J Nat Hist 16:315–334
Perera S, Molina MJ (2007) Plagas y enfermedades en el
cultivo ecolo
´gico de la platanera. In: Nogueroles C, Lı
´b-
ano J (eds) El cultivo ecolo
´gico de la platanera en
Canarias. Gabinete de Proyectos Agroecolo
´gicos S.L.,
Santa Cruz de Tenerife, pp 70–118
Pinto JD (1999) Systematics of the North American species of
Trichogramma Westwood (Hymenoptera: Trichogramm-
matidae). Mem Entomol Soc Wash 22:1–287
Pintureau B (2008) Les espe
`ces europe
´ennes des tricho-
grammes. Libro Veritas, Cergy-Pontoise, pp 1–95
Pintureau B, Babault M (1988) Syste
´matique des espe
`ces
africaines des genres Trichogramma Westwood et
Trichogrammatoidea Girault (Hym. Trichogrammmati-
dae). Trichogramma and other egg parasites. In: Voegele
´
J, Waage J, van Lenteren JC (eds) Second international
symposium, Guangzhou, China, November 10–15, 1986,
pp 97–120
Polaszek A (2010) Biodiversity and host associations of
Trichogramma in Eurasia, Chapter 9, pp. 237–266. In:
Consoli FL, Parra JRP, Zucchi RA (eds) Egg parasitoids
in agroecosystems with emphasis on Trichogramma.
Progress in Biological Control, vol 9. Springer, New
York, pp 1–479
Roditakis E, Papachristos D, Roditakis NE (2010) Current
status of the tomato leafminer Tuta absoluta in Greece.
EPPO Bull 40(1):163–166
Silva IMMS, Honda J, van Kan F, Hu J, Neto L, Pintureau B,
Stouthamer R (1999) Molecular differentiation of five
Trichogramma species occurring in Portugal. Biol Control
16(2):177–184
Siqueira HAA, Guedes RNC, Picanco MC (2000) Insecticide
resistance in populations of Tuta absoluta (Lep.: Gele-
chiidae). Agric For Entomol 2:147–153
Stouthamer R, Hu J, van Kan FJPM, Platner GR, Pinto JD
(1999) The utility of Internally Transcribed Spacer 2 DNA
sequences of the nuclear ribosomal gene for distinguish-
ing sibling species of Trichogramma (Hymenoptera:
Trichogrammatidae). BioControl 43:421–440
Sumer F, Tuncbilek AS, Oztemiz S, Pintureau B, Rugman-
Jones P, Stouthamer R (2009) A molecular key to the
common species of Trichogramma of the Mediterranean
region. BioControl 54(5):617–624
Thomson L, Rundle BJ, Carew ME, Hoffmann AA (2003)
Identification and characterization of Trichogramma spe-
cies from south-eastern using the internal transcribed
spacer 2 (ITS2) region of the ribosomal gene complex.
Entomol Exp Appl 106:235–240
Urbaneja A, Vercher R, Navarro V, Garcı
´a-Marı
´F, Porcuna JL
(2007) La polilla del tomate, Tuta absoluta. Phytoma Esp
194:16–23
Urbaneja A, Monton H, Molla O, Beitia F (2008) Suitability of
the Tomato Borer Tuta absoluta as prey for Macrolophus
pygmaeus and Nesidocoris tenuis. J Appl Entomol
133:292–296
Uygun N, Ozgur F (1980) Identification of pests of greenhouse
vegetables in the Icel and Adana regions, and the
effects of endosulfan smoke tablets and pirimicarb on
Myzus persicae (Sulz.). Turk Bitk Koruma Derg 4(3):
185–192
Vacante V, Cocuzza G, Pucci C, De Clercq P (1996) Note su
Podisus maculiventris (Say) e P. nigrispinus (Dallas)
(Heteroptera, Pentatomidae). Inf Fitopatol 46:11–16
Veire M van de (1993) First observations in glasshouse sweet
peppers in Belgium and laboratory rearing of the parasitic
wasp Eulophus pennicornis (Hym.: Eulophidae). Entom-
ophaga 38(1):61–62
Villas-Boas GL, Franca FH (1996) Use of the parasitoid
Trichogramma pretiosum for control of Brazilian tomato
pinworm in tomato grown in the greenhouse. Hortic Bras
14:223–225
Vila E, Parra A, Gallego JR, Ferna
´ndez FJ, Cabello T (2010)
Biological control of the South American Tomato Pin-
worm, Tuta absoluta with releases of Trichogramma on
different tomato crop cycles in Spain. In: IXth European
congress of entomology, 22–27 August, Budapest, Hungary
Vos R de, Rutten ALM (1995) Migrating Lepidoptera in 1992
(fifty-third report). Entomol Ber 55(3):37–46
Zandigiacomo P (1990) The principal pests of soyabean in
north-eastern Italy in 1989. Inf Fitopatol 40(8):55–58
Molecular and morphological diagnoses of five species of Trichogramma 35
123
View publication statsView publication stats