rte

JHR 68: 37-48 (2019) JOURNAL OF | *rrievet cvenace oar
doi: |0.3897/jhr.68.30372 RESEARCH ARTICLE ME Hymenoptera

http://jhr.pensoft.net The Inarrational Society of Hymenoptersts, RESEARCH

The attraction of Tremex apicalis (Hymenoptera,
Siricidae, Tremecinae) and its parasitoid [balia japonica
(Hymenoptera, Ibaliidae) to the fungus Cerrena unicolor

Kazumu Kuramitsu', Teruhito Ishihara’, Aki Sugita'’, Thitaree Yooboon*®,
Barry Lustig®, Yuko Matsumori’, Hideo Yamada®, Natsuko Kinoshita!

| Faculty of Life and Environmental Sciences, University of Isukuba, Tennodai 1-1-1, Tsukuba, Ibaraki 305-
8572, Japan 2. School of BioSciences, The University of Melbourne, Melbourne, Victoria 3010, Australia
3 Department of Evolutionary Studies of Biosystems, School of Advanced Sciences, SOKENDAI (The Graduate
University for Advanced Studies), Shonan Village, Hayama, Kanagawa 240-0193, Japan 4 Graduate School of
Life and Environmental Sciences, University of Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki 305-8572, Japan
5 Department of Zoology, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand 6 Cormorant
Group, Pittsburgh, Pennsylvania, 15253, USA 7 College of Life and Environmental Sciences, University of
Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki 305-8572, Japan 8 Tsukuba Experimental Forest, Mountain
Science Center, University of Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki 305- 8577, Japan

Corresponding authors: Kazumu Kuramitsu (kuramitsu.kazumu.ws@alumni.tsukuba.ac.jp);

Natsuko Kinoshita (kinoshita.natsuko.gf@u.tsukuba.ac.jp)

Academic editor: /. Yoder | Received 6 October 2018 | Accepted 8 January 2019 | Published 25 February 2019
http://zoobank. ore/C1 1LESAAB-76C5-4709-813F-D48D5A9D2F1E

Citation: Kuramitsu K, Ishihara T, Sugita A, Yooboon T, Lustig B, Matsumori Y, Yamada H, Kinoshita N (2019) The
attraction of Tremex apicalis (Hymenoptera, Siricidae, Tremecinae) and its parasitoid [balia japonica (Hymenoptera,
Ibaliidae) to the fungus Cerrena unicolor. Journal of Hymenoptera Research 68: 37-48. https://doi.org/10.3897/
jhr.68.30372

Abstract
Woodwasps (Hymenoptera: Siricidae) are saproxylic insects and a common forest pest. Siricid woodwasps
are classified into two subfamilies: Siricinae and Tremecinae. All known symbiotic fungi of Siricinae are in
the genus Amylostereum Boidin while some species of Tremecinae have been observed to have a relation-
ship with the fungus Cerrena unicolor (Bull.) Murrill. Previous studies about the host searching behavior
of woodwasps and their parasitoids have focused primarily on the subfamily Siricinae.

We analyzed the role of C. unicolor volatiles on the host searching behavior of Tremex apicalis Mat-

sumura (Hymenoptera: Siricidae: Tremecinae) and its parasitoid [balia (Tremibalia) japonica Matsumura

Copyright Kazumu Kuramitsu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC
BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

38 Kazumu Kuramitsu et al. / Journal of Hymenoptera Research 68: 37-48 (2019)

(Hymenoptera: Ibaliidae). The results of an olfactory response experiment indicated that the females of 7’
apicalis and its parasitoid find their respective hosts using volatiles from C. unicolor. Using DNA barcode,
we identified basidiocarps on the trees infested with 7’ apicalis. The basidiocarps were all white-rot fungi
that cause sapwood decay, including C. unicolor. Two additional species that we identified belonged to
genera closely related to C. unicolor.

Woodwasp species are known to carry symbiotic fungi in a pair of specialized sacs called mycangia.
Notably we found that mycangia-like structures were absent in the abdomens of 7. apicalis females. To the
best of our knowledge, Xeris spectrum (Linnaeus) (Hymenoptera: Siricidae) is the only reported example
of woodwasp species that do not contain symbiotic fungi in their bodies.

Our results suggested that: (1) 7’ apicalis females search for host wood that is already infected with
sapwood decaying fungus using volatile compounds; (2) T’ apicalis’ female parasitoid also uses volatile

compounds from fungus to locate wood that is infested with its potential host.

Keywords

Woodwasp, horntail, host searching, mycangia, saproxylic insect, Tremibalia, Y-tube

Introduction

Saproxylic insects, like woodwasps in the family Siricidae and their parasitoids, locate
suitable host wood/host insect-infested wood in their environment to increase their
reproductive success (Feldhaar and Schauer 2018; Hilszczariski 2018; Ulyshen and
Sobotnik 2018). Siricidae consists of two subfamilies: Siricinae and Tremecinae. Sir-
icinae infest coniferous trees and Tremecinae infest broad-leaved trees (Morgan 1968;
Schiff et al. 2012). Like most saproxylic insects, woodwasps are unable to digest struc-
tural polysaccharides such as lignocellulosic compounds because they lack the neces-
sary cellulolytic enzymes (Fukuda and Hijii 1997; Slippers et al. 2012).

Many woodwasp species carry fungal symbionts in their mycangia. Symbiotic
fungi are transferred to host wood during oviposition and hatched larvae feed on the
fungus-infected wood. To date, all known symbiotic fungi of Siricinae are in the genus
Amylostereum Boidin. A limited example of Tremecinae woodwasps demonstrates that
this subfamily is associated with fungus Cerrena unicolor (Bull.) Murrill (Stillwell 1967;
Tabata and Abe 1995; Pazoutova and Srutka 2007; Kuramitsu et al. 2016).

Siricinae woodwasps use semiochemicals emitted by trees to locate host wood. For
example, Siricinae species of Sirex, Urocerus and Xeris (Hymenoptera: Siricidae: Sirici-
nae) are attracted to monoterpene hydrocarbons from host pine trees (Sato and Maeto
2006; Matsumoto and Sato 2007; Coyle et al. 2012; Matsumoto and Sato 2012; Er-
bilgin et al. 2017; Kties et al. 2018). European woodwasp, Sirex noctilio Fabricius, is
similarly attracted to the volatiles from its fungal symbiont, Amylostereum areolatum
(Chaillet ex Fr.) Boidin (Fernandez Ajo et al. 2015).

Like S. noctilio, egg-larval or larval endoparasitoids of Siricinae, [balia (Ibalia) spp.
(Hymenoptera: Ibaliidae) locate their hosts using the symbiotic fungi volatiles of their
hosts (Madden 1968; Spradbery 1974; Martinez et al. 2006; Jofré et al. 2016; Kiies
et al. 2018; Table 1). For example, /balia (.) leucospoides, a parasitoid of woodwasp S.
noctilio, is attracted to the volatiles from their fungal symbiont (Martinez et al. 2006;

The attraction of Tremex apicalis... ao

Table |. Relationship of Ibaliid parasitoids and their host.

Ibaliid parasitoids Host Host trees of host Symbiotic fungi of References’
woodwasps woodwasps host woodwasps
Ibalia (Ibalia) spp. Siricinae Coniferous trees Amylostereum spp. 1, 2, 3, 4,5
Ibalia (Tremibalia) spp. "Tremecinae Broad-leaved trees Cerrena unicolor 1, 2, 3, 4, 6, 7, 8

* 1 Nordlander and Liu (1994), 2 Choi et al. (2013), 3 Morgan (1968), 4 Schiff et al. (2012), 5 Tabata et al. (2012),
6 Stillwell (1967), 7 Pazoutova and Srutka (2007), 8 Tabata and Abe (1995).

Pietrantuono et al. 2012). Parasitoid wasps exhibit an antennal palpating and oviposi-
tor probing response to discs of fungus impregnated agar (Spradbery 1974).

Information about the host wood/host insect searching behavior of Tremecinae
and their parasitoids, /balia (Tremibalia) spp., is limited. The information available fo-
cuses on the attraction of Tremex columba (Linnaeus) (Hymenoptera: Siricidae: Trem-
ecinae) to the wood volatile «/8-pinen (Coyle et al. 2012).

C. unicolor is the only known fungal symbiont of Tremecinae based on previous stud-
ies of Tremex spp. (Hymenoptera: Siricidae: Tremecinae) (Stillwell 1967; Tabata and Abe
1995; Pazoutova and Srutka 2007; Table 1). Basidiocarps which had the morphology of
C. unicolor were present on wood infested by Tremex apicalis Matsumura (Kuramitsu et al.
2016). Whether or not C. unicolor is a symbiotic fungi of 7- apicalis is not yet determined.
To clarify the interaction between 7’ apicalis and C. unicolor: (1) we dissected the abdo-
mens of 7. apicalis females to isolate their mycangia, (2) analyzed the role of C. unicolor
volatiles on T’ apicalis’ behavior, (3) identified basidiocarps on T’ apicalis infested trees.

Also, we hypothesized that lbalia (T:) japonica Matsumura, a parasitoid of T’ api-
calis, uses volatiles from C. unicolor to locate trees with potential host woodwasps. To
test this hypothesis, we investigated the role of fungus volatiles on the host searching
behavior of parasitoid /. japonica under laboratory conditions.

Materials and methods

Site of study and host trees

Our field survey and insect collection was conducted at Tsukuba Experimental Forest,
University of Tsukuba (36°07'10"N; 140°05'50"E (DMS), ca. 25 m a.s.l.), Tsukuba, Iba-
raki Prefecture, Honshu, Japan. We found four 7’ apicalis infested trees belonging to differ-
ent families from 2016 to 2018 (Table 2). All 7’ apicalis, parasitoids and basidiocarps used
for dissection, observation and behavioral experiments were collected from these four trees.

Observations on the abdominal organs of female TZ. apicalis

Twenty 7’ apicalis females were collected from the aforementioned trees (Table 2). 77
apicalis females were killed using ethyl acetate. The dissection method was based on

40 Kazumu Kuramitsu et al. / Journal of Hymenoptera Research 68: 37-48 (2019)

Table 2. Host trees from which woodwasps, parasitoids and basidiocarps were collected.

Tree Tree species Diameter at Emergence of T. apicalis/ I. japonica Year of basidiocarp
no. breast height 2016 2017 2018 collection
i Swida macrophylla (Wall.) 40 cm yes / yes yes / no no / no 2016
(Cornales: Cornaceae)
2 Euptelea polyandra Sieb. 19 cm no / no yes / yes yes / yes 2018
et Zucc. (Ranunculales:
Eupteleaceae)
3 Fraxinus spaethiana Lingelsh 23 cm _ yes / yes no/ no no fungi

(Scrophulariales: Oleaceae)
4 Magnolia liliiflora Dest. 44 cm _ yes / yes yes / yes 2018
(Magnoliales: Magnoliaceae)

* This tree was the same tree that Kuramitsu et al. (2016) studied.
** These trees were not observed in 2016.

previous studies (Thomsen and Harding 2011; Li et al. 2015). The abdomen was re-
moved using micro scissors under a stereomicroscope (Leica MZ12). The dorsal plates
were removed from the abdomen. A female Tremex longicollis Konow, which is known
to have the mycangia (Tabata and Abe 1995), was also dissected using the same meth-
od. The 7’ longicollis female was collected in Yokohama, Japan on November 12, 2017.

Extracting DNA from basidiocarps and PCR amplification of fungal ribosomal DNA

Collected basidiocarp surfaces were removed to avoid potential contamination. The
samples were then ground into a fine powder using a pestle, mortar and liquid nitro-
gen. Fungal DNA was extracted from the powdered samples by using DNeasy Plant
Mini Kit (QIAGEN) following the manufacturer’s instructions. Each extracted DNA
sample was used as a PCR template to amplify an Internal Transcribed Spacer (ITS)
region by using KOD FX Neo (Toyobo) following the manufacturer's instructions.
Reactions were performed with 25 yl mixture containing KOD FX Neo, 2xbuffer for
KOD FX Neo, 2 ».M dNTP, 0.3 uM of each primer. Primers used to amplify fungal
ITS region were ITS4 (5’°-TCCTCCGCT TATTGATATGC-3’) and ITS5 (5’-GGAA-
GTAAAAGTCGTAACAAGG-3’) (White et al. 1990). PCR reactions were performed
in the following order: 94 °C for 2 minutes, followed by 40 cycles of 98 °C for 10
seconds, 50 °C for 30 seconds, and 68 °C for 90 seconds. PCR products around 650
baseparis were purified using a QlAquick Gel Extraction Kit (QIAGEN).

Sequencing and molecular identification

Sequence reactions were performed with BigDye Terminator v3.1 (Thermo Fisher Sci-
entific) followed by purification using BigDye Xterminator (Thermo Fisher Scientific).
The Sanger method was applied to determine DNA sequence of ITS region using

The attraction of Tremex apicalis... 4]

Applied Biosystems 3130 (Gene Research Center, University of Tsukuba) and com-
mercial sequencing services (Macrogen Japan and Eurofins Genomics). Fungi species
were identified using the UNITE database (http://unite.ut.ee) (Koljalg et al. 2013).
ITS sequences were deposited to Genbank (accession numbers are: Cerrena unicolor,

MH645754; Daedaleopsis confragosa, MH645755; Trametes hirsuta, MH645756).

Fungal culture disc preparation

Potato Dextrose Agar (PDA) medium (Nissui) was prepared by following the manu-
facturer’s protocol. C. unicolor was obtained from the Genebank Project (National
Agricultural Research Organization). For behavioral experiments, fungal cultures were
inoculated with a PDA medium in a 9-cm petri dish for two weeks at 25 °C.

Olfactory responses of T. apicalis and its parasitoid I. japonica

To obtain newly emerged T’ apicalis and I. japonica, we cut down woodwasp infested
E. polyandra (Table 2, tree no. 2) and M. liliiflora (Table 2, tree no. 4) on November
11 and October 4, 2017, respectively. The wood was stored outside until May 2018.
Newly emerged T apicalis and I. japonica were collected upon emergence. Live adults
of each species was stored in plastic containers (16 x 28 x 17 cm). All insects were al-
lowed to mate and feed on a solution of sugar and water (30% w/w) for 24—48 hours
before behavioral experiments. The olfactory preference of 7’ apicalis and I. japonica
was examined with a Y-tube bioassay (Fig. 1a) in the laboratory (25 °C = 1.1 °C). Arms
of a glass Y-tube olfactometer (common arm 20 cm, arms 20 cm, diameter 3 cm) were
connected to glass jars (17 cm x 12.5 cm) respectively. Each glass jar was connected
to an activated charcoal filter. The Y-tube olfactometer was connected to an electric
vacuum pump (KNE, Germany) through a flow meter. The arm side of the Y-tube ol-
factometer was inclined upward at 5-degrees using a metal stand (Fig. 1b). The airflow
from outside passed through the charcoal, the glass jar and into the olfactometer. The
flow rate of the pump was set at 1.5 L/min.

A single male or female woodwasp or parasitoid was introduced into the start-
ing point of the Y-tube olfactometer. Its behavior was observed for maximum of 15
minutes. We recorded the choice of the wasp if it reached the finish line. If it did not
make a choice within 15 minutes, the trial was recorded as, “no choice.” A total 43
T. apicalis (20 females and 23 males) and 57 I. japonica (25 females and 32 males)
were tested. To avoid bias in the experimental setup, the positions of the two odors
sources were exchanged after testing five woodwasps or parasitoids. Odor sources were
renewed after testing five woodwasps or parasitoids. Using a binomial test, we deter-
mined the preference of both 7’ apicalis and I. japonica between volatiles from fungal
disks and control disks.

42 Kazumu Kuramitsu et al. / Journal of Hymenoptera Research 68: 37-48 (2019)

(a)

Activated charcoal

White-washed
| plastic wall

Y-tube olfactometer

Plastic board

Flow meter

Figure |. Set-up of the Y-tube olfactometer used to test 7’ apicalis and I. japonica attraction to volatiles
from a fungal disc of C. unicolor in a top view (a) and in a side view (b) of the Y-tube olfactometer.

Results

Female T. apicalis abdominal organs

Abdomens of the female 7’ apicalis had ovaries that contained on average 96 eggs (Fig. 2a).
A pair of mucus glands, which looked like large whitish sacs, was highly developed (Fig. 2a).
A single mucus reservoir connected to the basement of the ovipositor was detected under
the mucus glands (Fig 2b). The reservoir contained a sticky transparent fluid.

The attraction of Tremex apicalis... 43

(a) (b)

Figure 2. Abdominal organs of female woodwasps. a Ventral view of the dissected abdomen of T' apicalis.
Typical internal organs, ovaries and mucus glands were easily detected. b Upper left diagonal view of the
dissected abdomen of 7’ apicalis. A mucus reservoir was detected under the mucus glands. ¢ Comparative
view of dissected region of abdomens close to the ovipositor. In 7’ longicollis, the mycangia are recognized as
greyish balls located close to the basement of ovipositor. There were no such sac-like structures in T7 apicalis.

Abbreviations; my: mycangia, mr: mucus reservoir, mg: mucus gland, ov: ovary. Scale bar: 2 mm.

We could not locate mycangia-like structures in the abdomen of the female 7’ api-
calis while other anatomical features were nearly identical to other woodwasps. There
were no visible sac-like structures located behind the base of the ovipositor where 7.
longicollis has clearly identifiable mycangia (Fig. 2c).

44 Kazumu Kuramitsu et al. / Journal of Hymenoptera Research 68: 37-48 (2019)

OControl |&Fungal culture disc

P<0.05
Tremex apicalis female |
(n = 17)
P = 0.774
(n = 12)
P<0.05
Ibalia japonica female |
(n = 21)
P = 0.824
Ibalia japonica male ——si—
(n = 20)
60 40 20 0 20 40 60 80

Choice (%)

Figure 3. Percent choice by 7 apicalis and I. japonica in arms of Y-tube olfactometer with the volatiles

from PDA discs (control) vs. fungal culture discs.

Identification of basidiocarps on woodwasp infested trees

We performed DNA barcoding for basidiocarps found on T’ apicalis infested trees using
ITS region. The results showed that the ITS region from basidiocarps on: (1) S. macrophyl-
la had a 99.67% match with C. unicolor; (2) E. polyandra had a 99.81% match to Dae-
daleopsis confragosa (Bolton) J.Schrét.; and (3) M. Liliiflora was identical to Trametes hirsuta
(Wulfen) Lloyd. All the fungus species we identified belong to the family Polyporaceae.

Olfactory responses of T. apicalis and its parasitoid I. japonica

In the Y-tube bioassay, 85.0% (n = 20) of 7 apicalis females, 52.2% (n = 23) of its
males, 84.0% (n = 25) of L. japonica females and 62.5 % (n = 32) of males chose be-
tween the volatiles from fungal and control disks. Females of 7’ apicalis woodwasps
(76.5 %, n= 17, P< 0.05) and L. japonica (76.2%, n = 21, P< 0.05) preferred vola-
tiles from the fungal disk to the control disks (Fig. 3). In contrast, males of both
LT. apicalis (41.7%, n = 12, P= 0.77) and I. japonica (55.0%, n = 20, P = 0.82) did

not display a statistically significant preference for either fungal or control disks.

Discussion

T. apicalis females without mycangia feed opportunistically on rotten wood

Subfamily Siricinae has a close relationship with the genus Amylostereum (Schiff et
al. 2012; Tabata et al. 2012). For subfamily Tremecinae, C. unicolor is only known

The attraction of Tremex apicalis... 45

symbiotic fungus for many Tremex woodwasps (Stillwell 1967; Tabata and Abe 1995;
Pazoutova and Srutka 2007). Basidiocarps on 7. apicalis infested large-leaf dogwood
tree (Swida macrophylla) were C. unicolor. Notably, we could not identify any apparent
mycangia-like structures inside the female 7’ apicalis (Fig. 2).

Basidiocarps on other 7’ apicalis infested trees were members of the family Polypo-
raceae, inclusive of C. unicolor. These species are white-rot fungi that cause sap wood
decay (Enebak and Blanchette 1989; Stajié et al. 2017; Racko et al. 2018). Our obser-
vations suggest that 7’ apicalis, which lacks identifiable mycangia, inhabits host wood
that is already infected by wood decaying fungi.

Relationship between C. unicolor and T. apicalis

The female 7’ apicalis preference for C. unicolor suggests that it uses volatiles from the fun-
gus to locate suitable host wood. ‘This strategy would be similar to female Xeris spectrum
(Hymenoptera: Siricidae), whose mycangia is also absent and who use the odor from fun-
gi Amylostereum to locate host wood (Fukada and Hijii 1997; Matsumoto and Sato 2012).

While mycangia carrying woodwasps Sirex nitobei Matsumura and Uvocerus
japonicus Smith form specific relationships with a particular Amylostereum fungus, my-
cangia-less X. spectrum can utilize more than one particular species. For example, the
Amylostereum fungi species has a well-documented relationship with Sirex nitobei and
Urocerus japonicas respectively (Fukuda and Hijii 1997).

It is possible that 7! apicalis employs similar strategies that take advantage of their
lack of mycongia. This may provide woodwasps without mycongia with an evolution-
ary advantage that increases their overall chances for reproduction and survival.

Parasitoid strategies

The Siricinae parasitoid 1. leucospoides locates its host using volatile cues from symbiotic
fungi (Martinez et al. 2006; Pietrantuono et al. 2012). In our study, we demonstrated
that /. japonica is attracted to volatile compounds from C. unicolor even though this
fungus is not a symbiont carried in 7’ apicalis. In contrast, 1. japonica’s potential host, 7.
longicollis, has symbiotic relationship with C. unicolor (Tabata and Abe 1995; Watanabe
et al. 2018). In order to locate potential hosts, /balia parasitoids seem to exploit olfacto-
ry cues not only from symbiotic fungi but also fungi species that live outside of the host.

To the best of our knowledge, this is the first report that connects white rot wood de-
caying fungus, Iremecinae woodwasp species and its parasitoid via volatile compounds.

Acknowledgments

We are grateful to the members of staff at the Experimental Forest Station, University
of Tsukuba, Japan for their permission to conduct this study. We are also grateful to

46 Kazumu Kuramitsu et al. / Journal of Hymenoptera Research 68: 37-48 (2019)

Dr. Yooichi Kainoh (University of Tsukuba), Mr. Atsuya Kosaki (Yokohama, Japan)
and Dr. Rikio Matsumoto (Osaka Museum of Natural History) for their ongoing en-
couragement and practical assistance. We are indebted to Dr. Yuho Ando (Forestry and
Forest Products Research Institute), Dr. Kimiyo Matsukura (Tohoku University), and
Dr. Izumi Okane (University of Tsukuba) for their technical expertise on mycology.
We also thank Genebank Project (National Agricultural Research Organization) for
providing a strain of the fungus Cerrena unicolor. This research was supported, in part,
by Tomizawa Jun-ichi & Keiko Fund of Molecular Biology Society of Japan for Young

Scientists and Canon Foundation Grant “Pursuit of Ideal.”

References

Choi WY, Lee JW, Suh KI (2013) Taxonomic review of the family Ibaliidae (Cynip-
oidea: Hymenoptera) from Korea. Entomological Research 43: 135-141. https://doi.
org/10.1111/1748-5967.12015

Coyle DR, Pfammatter JA, Journey AM, Pahs TL, Cervenka VJ, Koch RL (2012) Community
composition and phenology of native Siricidae (Hymenoptera) attracted to semiochemi-
cals in Minnesota. Environtal Entomology 41: 91-97. https://doi.org/10.1603/EN11192

Enebak SA, Blanchette RA (1989) Canker formation and decay in sugar maple and paper birch
infected by Cerrena unicolor. Canadian Journal of Forest Research 19: 225-231. https://
doi.org/10.1139/x89-031

Erbilgin N, Stein JD, Acciavatti RE, Gillette NE, Mori SR, Bischel K, Cale JA, Carvalho CR,
Wood DL (2017) A blend of ethanol and (-)-«-pinene were highly attractive to native
siricid woodwasps (Siricidae, Siricinae) infesting conifers of the Sierra Nevada and the Al-
legheny Mountains. Journal of Chemical Ecology 43: 172-179. https://doi.org/10.1007/
s10886-016-0803-y

Feldhaar H, Schauer B (2018) Dispersal of saproxylic insects. In: Ulyshen MD (Ed.) Saproxylic
insects, zoological monographs 1. Springer, Cham, 515-546. https://doi.org/10.1007/978-
3-319-75937-1_15

Fernandez Ajo AA, Martinez AS, Villacide JM, Corley JC (2015) Behavioural response of the
woodwasp Sirex noctilio to volatile emissions of its fungal symbiont. Journal of Applied
Entomology 139: 654-659. https://doi.org/10.1111/jen.12211

Fukuda H, Hijii N (1997) Reproductive strategy of a woodwasp with no fungal symbionts, Xeris
spectrum (Hymenoptera: Siricidae). Oecologia 112: 551-556. https://doi.org/10.1007/
5004420050344

Hilszczanski J (2018) Ecology, diversity and conservation of saproxylic hymenopteran para-
sitoids. In: Ulyshen MD (Ed.) Saproxylic Insects, Zoological Monographs 1. Springer,
Cham, 193-216. https://doi.org/10.1007/978-3-319-75937-1_6

Jofré N, Pildain MB, Cirigliano AM, Cabrera GM, Corley JC, Martinez AS (2016) Host se-
lection by lbalia leucospoides based on temporal variations of volatiles from the hosts’ fun-
gal symbiont. Journal of Applied Entomology 140: 736-743. https://doi.org/10.1111/
jen.12313

The attraction of Tremex apicalis... 47

Koljalg U, Nilsson RH, Abarenkov K, et al. (2013) Towards a unified paradigm for sequence-
based identification of Fungi. Molecular Ecology 22: 5271-5277. https://doi.org/10.1111/
mec.12481

Kiies U, Khonsuntia W, Subba S, Dornte B (2018) Volatiles in communication of Agaricomy-
cetes. In: Anke T, Schiiffler A (Eds) Physiology and Genetics The Mycota (A Comprehen-
sive Treatise on Fungi as Experimental Systems for Basic and Applied Research), vol 15.
Springer, Cham, 149-212. https://doi.org/10.1007/978-3-319-71740-1_6

Kuramitsu K, Kosaki A, Ishihara T, Yamada H, Watanabe K (2016) Infestation of the wood-
wasp Tremex apicalis Matsumura (Hymenoptera, Siricidae) on the large-leaf dogwood
Swida macrophylla (Wall.) with biological notes on its parasitoid wasps. Journal of Hyme-
noptera Research 52: 71—79. https://doi.org/10.3897/jhr.52.10060

Li D, Shi J, Lu M, Ren L, Zhen C, Luo Y (2015) Detection and identification of the invasive
Sirex noctilio (Hymenoptera: Siricidae) fungal symbiont, Amylostereum areolatum (Russu-
lales: Amylostereaceae), in China and the stimulating effect of insect venom on laccase
production by A. areolatum YQLO3. Journal of Economic Entomology 108: 1136-1147.
https://doi.org/10.1093/jee/tov072

Madden JL (1968) Behavioural responses of parasites to the symbiotic fungus associated with
Sirex noctilio F. Nature 218: 189-190. https://doi.org/10.1038/218189a0

Martinez AS, Fernandez-Arhex V, Corley JC (2006) Chemical information from the fungus Am-
ylostereum areolatum and host-foraging behaviour in the parasitoid [balia leucospoides. Phys-
iological Entomology 31: 336-340. https://doi.org/10.1111/j.1365-3032.2006.00523.x

Matsumoto T, Sato S (2007) An olfactometer testing the olfactory response of the Japanese
horntail Urocerus japonicus to volatiles. Journal of the Japanese Forest Society 89: 135-137.
https://doi.org/10.4005/jjfs.89.135 [In Japanese with English summary]

Matsumoto T, Sato S (2012) Differential responses to «-pinene of two horntail wasps, Urocerus
antennatus and Xeris spectrum (Hymenoptera: Siricidae). Bulletin of the Forestry and Forest
Products Research Institute 11: 51-55.

Morgan D (1968) Bionomics of Siricidae. Annual Review of Entomology 13: 239-256.
https://doi.org/10.1146/annurev.en.13.010168.001323

Nordlander G, Liu Z (1994) Review of the family Ibaliidae (Hymenoptera: Cynipoidea) with
keys to genera and species of the World. Insect Systematics & Evolution 25: 377-392.
https://doi.org/10.1163/187631294X00153

Pazoutova S, Srutka P (2007) Symbiotic relationship between Cerrena unicolor and the horntail
Tremex fuscicornis recorded in the Czech Republic. Czech Mycology 59: 83-90.

Pietrantuono AL, Fernandez-Arhex V, Jofré N, Corley JC (2012) Food and host searching
decisions made by Jbalia leucospoides (Hymenoptera: Ibaliidae), a parasitoid of Sirex
noctilio (Hymenoptera:Siricidae). Journal of Insect Behavior 25: 320-327. https://doi.
org/10.1007/s10905-011-9301-9

Ratko V, Misikova O, Hlavaé P, Dedkova V (2018). Can bark stripping cause red heartwood
formation in beech stems? iForest 11: 251-258. https://doi.org/10.3832/ifor2147-011

Sato S, Maeto K (2006) Attraction of female Japanese horntail Urocerus japonicus (Hymenop-
tera: Siricidae) to a-pinene. Applied Entomology and Zoology 41: 317-323. https://doi.
org/10.1303/aez.2006.317

48 Kazumu Kuramitsu et al. / Journal of Hymenoptera Research 68: 37-48 (2019)

Schiff NM, Goulet H, Smith DR, Boudreault C, Wilson AD, SchefHler BE (2012) Siricidae
(Hymenoptera: Symphyta: Siricoidea) of the Western Hemisphere. Canadian Journal of
Arthropod Identification 21: 1-305. https://doi.org/10.3752/cjai.2012.21

Slippers B, de Groot P, Wingfield MJ (2012) The Sivex woodwasp and its fungal symbiont:
research and management of a worldwide invasive pest. Springer, New York. https://doi.
org/10.1007/978-94-007-1960-6

Spradbery J (1974) The responses of /balia species (Hymenoptera: Ibaliidae) to the fungal sym-
bionts of siricid woodwasp host. Journal of Entomology Series A, General Entomology 48:
217-222. https://doi.org/10.1111/j.1365-3032.1974.tb00058.x

Staji¢ M, Cilerdzié J, Gali¢ M, Ivanovié Z, Vukojevié J (2017) Lignocellulose degradation by
Daedaleopsis confragosa and D. tricolor. BioResources 12: 7195-7204.

Stillwell MA (1967) The pigeon Tremex, Tremex columba (Hymenoptera: Siricidae), in New
Brunswick. The Canadian Entomologist 99: 685-689. https://doi.org/10.4039/Ent99685-7

Tabata M, Abe Y (1995) Cerrena unicolor isolated from the mycangia of a horntail, Tremex
longicollis, in Kochi Prefecture, Japan. Mycoscience 36: 447—450. https://doi.org/10.1007/
BF02268630

Tabata M, Miyata H, Maeto K (2012) Siricid woodwasps and their fungal symbionts in Asia,
specifically those occurring in Japan. In: Slippers B, de Groot P, Wingfield MJ (Eds) The
Sirex Woodwasp and its Fungal Symbiont: Research and Management of a Worldwide In-
vasive Pest. Springer, New York, 95-102. https://doi.org/10.1007/978-94-007-1960-6_7

Thomsen IM, Harding S (2011) Fungal symbionts of siricid woodwasps: Isolation techniques
and identification. Forest Pathology 41: 325-333. https://doi.org/10.1111/j.1439-
0329.2010.00677.x

Ulyshen MD, Sobotnik J (2018) An introduction to the diversity, ecology, and conservation
of saproxylic insects. In: Ulyshen MD (Ed.) Saproxylic Insects, Zoological Monographs 1.
Springer, Cham, 1—47. https://doi.org/10.1007/978-3-319-75937-1

Watanabe K, Ito M, Fujie S, Shimizu S (2018) Information station of Parasitoid wasps. http://
himebati.jimdo.com/ [Accessed 17 July 2018]

White TJ, Bruns TD, Lee SB, Taylor JW (1990) Amplification and direct sequencing of fungal
ribosomal RNA Genes for phylogenetics. In: Innis MA, Gelfand HD, Sninsky JJ, White
TJ (Eds) PCR Protocols: A Guide to Methods and Applications. Academic Press, New
York, 315-322. https://doi.org/10.1016/B978-0-12-372180-8.50042-1