Dicyphus (Dicyphus) bolivari Lindberg, 1934

Dicyphus (Dicyphus) bolivari Lindberg, 1934 View in CoL

Dicyphus bolivari Lindberg, 1934: 12 View in CoL

= Dicyphus (Dicyphus) bolivari atlanticus Wagner, 1951: 29 View in CoL (syn. SAN- CHEZ & CASSIS 2018: 39). Confirmed synonymy.

= Dicyphus (Dicyphus) maroccanus Wagner, 1951: 19 View in CoL (syn. SANCHEZ & CASSIS 2018: 39). Confirmed synonymy.

= Dicyphus (Dicyphus) tamaninii Wagner, 1951: 16 View in CoL . New synonymy.

Type material examined. Dicyphus bolivari Lindberg, 1934 . HOLOTY- PE: SPAIN: J (brachypterous): ‘[S:a Morena. St:a Helena 4 ― 8 4 26 Lindberg] / [Coll. Lindberg] / [Spec. typ. Dicyphus bolivari Lindberg ] / [MUS ZOOL HELSINKI loan nr HE 06-44.’ ( MZHF)]. Specimen not dissected, the left paramere was visible in lateral view ( Figs 1A–B View Fig ) and we have opted not to take the risk of damaging it. – PARATYPES: CANARY ISLANDS: ♀ (macropterous) [GZ50300], Spain: Canary Islands, Tenerife, Santa Ursula 18.V.1947 Lindberg, Hakan leg. ( MZHF); J (macropterous) [GZ50301], Spain:Canary Islands,Tenerife,Santa Ursula 18.V.1947 Lindberg, Hakan leg. ( MZHF); ♀ (macropterous) [GZ50309], Spain: Canary Islands, Tenerife, Santa Cruz 4.IV.1949 Lindberg, Hakan leg. / Photographied 2022 Pekka Maninen ( MZHF); ♀ (macropterous) [GZ50313], Spain: Canary Islands, Tenerife, Santa Cruz 4.IV.1949 Lindberg, Hakan leg. ( MZHF); ♀ (macropterous) [GZ50314], Spain: Canary Islands, Tenerife, Santa Cruz 4.IV.1949 Lindberg, Hakan leg. ( MZHF); J (macropterous, dissected): Spain: Canary Islands, Tenerife, Santa Cruz 4.IV.1949 Lindberg, Hakan leg. ( Figs 1E–G View Fig ) ( MZHF); ♀ (macropterous, dissected) [GZ50316], Spain: Canary Islands, Tenerife, Santa Cruz 14.I.1949 Lindberg, Hakan leg. ( Figs 1H, J View Fig ) ( MZHF).

Dicyphus bolivari atlanticus Wagner, 1951 . PARATYPE: CANARY ISLANDS: ♀ (macropterous) [GZ50324], Spain: Canary Islands, Tenerife, Santa Cruz 4.IV.1949 Lindberg, Hakan leg. / Photographied 2022 Pekka Maninen. Paratype D. bolivari atlanticus E.Wagn / coll. Lindberg. ( MZHF).

Dicyphus tamaninii Wagner, 1951 . PARATYPES: CROATIA: ♀ (macropterous, dissected): [Dalmatien, Split 13.5.43 Novak leg. Hyoscyamus niger L] / [Paratypus Dicyphus tamaninii n.sp. E.Wagner det.Paratypoid Dicyphus tamaninii n.sp.] / [ZMH 838481] ( Figs 1I, K View Fig ) ( ZMUH); ♀ (macropterous, dissected), [Palagruža D. Novak 19.5.49 Hyoscyamus niger L] / [Paratypoid Dicyphus tamaninii n.sp. E. Wagner det.] / [ZMH 838485] ( ZMUH); J (brachypterous), [O Sušac D., 20.6.49, Novak leg. Hyoscyamus niger L] / [Paratypoid Dicyphus tamaninii n.sp. E. Wagner det.] / [ZMH 838488] ( ZMUH). Not dissected, the left paramere was visible in lateral view ( Figs 1C–D View Fig ) and we have opted not to take the risk of damaging it. ♀ (macropterous), [Dalmatien Split 19.5.43 Novak leg. Hyoscyamus niger L] / [Paratypus Dicyphus tamaninii n.sp. E.Wagner det. Paratypoid Dicyphus tamaninii n.sp.] / [ZMH] / [ZMH 838489] ( ZMUH).

Additional material identified as D. bolivari or D. tamaninii . More than 700 specimens of both sexes from the following collections were studied: AMPF, BABN, CBGP-INRAE, JSMF, MLGU, MNHN, MTDU and RMNH, from the following locations: Northern Cyprus (new record: Famagouste, 25 V 2007, AMPF), England, France including Corsica (departments: Aisne, Alpes-Maritimes, Aude, Bouches-du-Rhône, Charente, Cher, Côtes d’Armor, Drôme, Finistère, Haute-Garonne, Gironde, Hérault, Haute-Loire, Loire-Atlantique, Lozère, Manche, Marne, Orne, Pyrénées-Orientales, Paris, Sarthe,Tarn, Var,Vaucluse), Germany, Greece including Crete, Channel Islands: Guernsey (new record: Moulin Huet, 3.viii.2020, BABN), Lebanon, Monaco (new record: Glacis du Palais, 10/ 18.vii.2010, AMPF), Morocco, the Netherlands (from 52 different localities), Sardinia (new record: Sassari, road 292 near Villanova Monteleone, 1.vi.2001, AMPF), Spain, and Asian Turkey (new record: Burdur, road from Köseler to Kargi, bridge across the Aksu Cayi, 7.vii.1999, AMPF). Fresh material was collected from 2005 to 2022 and preserved in 96% ethanol for DNA analyses. All specimens sequenced are listed in Table 1.

Diagnosis. Adults. Males and females not very different. Macropterous and brachypterous forms known in both sexes. Habitus ( Fig. 2B View Fig ) similar to most Dicyphus species of the subgenus Dicyphus s. str., with no real distinguishing characters. Identification requires dissection of the male or female genitalia.

Size. Males: brachypterous 2.8–3.2 mm, macropterous 3.8–4.3 mm; females: brachypterous 2.9–3.5 mm; macropterous 4.0– 4.4 mm.

Habitus usually pale, macropterous sometimes with reddish tinge. Head marked by two broken dark lines, eyes brownish. Antennal segment I bicoloured, base and apex with reddish-brown annulations, II basally lightly stained with faded brown, apex more or less dark brown, III and IV greyish to dark. Pronotum: underside of mesothorax sometimes black. Legs pale with short dark pilosity, hind tibiae with long dark spines, widely spaced, last tarsal segment black. Hemelytron pale or iridescent with semi-erect pale brown pilosity, two dark spots anterior to cuneal fracture, apex of cuneus black. Male genitalia: left paramere apophysis elongate (245 to 367 μm from outer margin of base to tip of shaft); shaft clearly demarcated and weakly sinuate ( Figs 2A, C View Fig ); endosoma with 2 to 13 small sclerites ( Fig. 2D View Fig ). Female genitalia: genital chamber characterised by cordiform shape, sometimes simply rounded, with edge of the sac folded into very characteristic accordion shape. Oviducts are swollen at base, well separated and generally directed downward ( Figs 1J–K View Fig ).

Morphological variability. External characters. We have studied more than 700 specimens identified as D. bolivari or D. tamaninii and sometimes both depending on the specialist responsible for the identification, of which 103 males and 55 females were dissected. We concur with SANCHEZ & CASSIS (2018) that the body size and external characters of D. bolivari and D. tamaninii overlap strongly and we did not find any morphological characters such as size, proportions of different appendages, segments, head, pronotum etc., nor colouration, wing polymorphism that would justify a separation into two or more taxa.

Male genitalia. SANCHEZ & CASSIS (2018) continue their diagnosis of D. tamininii versus D. bolivari by “species separation is based on differences in the male genitalia, especially the size of the left paramere and the number of endosomal lobal sclerites”. In fact, the shape of the left paramere does not vary very much between the two species. The differences reported by WAGNER (1951, 1974) and WAGNER & WEBER (1964) can be interpreted as different orientations of the paramere or, in the best case, as intra-specific variability (e.g. Figs 2A and 2C View Fig ). SANCHEZ & CASSIS (2018) measured the apophysis from “outer margin of the base to tip of shaft” and gave the following measurement ranges to distinguish the two species: 440–450 μm for D. bolivari and 530–600 μm for D. tamaninii . We estimated these measurements on the holotype of D. bolivari (a brachypterous male not dissected) and the paratype ZMH838488 (also a brachypterous male) and found 228 μm for the holotype of bolivari and 312 μm for the paratype of tamaninii which is a larger specimen ( Figs 1A–D View Fig ). We also measured the shaft of apophysis of 20 males identified as D. bolivari / tamaninii . It varies between 245 and 367 μm. Obviously, SANCHEZ & CASSIS (2018) gave measurements that do not correspond to what they have defined in their material and methods ( SANCHEZ & CASSIS 2018: 343: fig. 9f) and used in their key. Another character given by these authors is the number of sclerites of the endosoma: 2–5 for D. tamaninii ; 8–12 (rarely 6–7) for D. bolivari . These sclerites are easy to observe ( Fig. 2D View Fig ) but much more difficult to count, especially when the penis is not inflated. We managed to evaluate the number of endosomal sclerites in 35 males and found that the number of sclerites varied from 4 to 13. We noticed that we could not verify that a low number of sclerites (2–5) was associated with a long apophysis (for D. tamaninii ) and a high number of sclerites (8-12) was associated with a short apophysis (for D. bolivari ). The apophysis of the male with 4 sclerites measured 275 μm, of that with 13 sclerites 347 μm. Under these conditions it is not possible to distinguish the two species, nor to use SANCHEZ & CAS- SIS’ s (2018) key. There is an intraspecific variation in the male genitalia but none of the criteria used by the authors allow the delimitation of two or more taxa.

Female genitalia. The female genitalia of the subgenus Dicyphus have never been studied. They will be described and illustrated in another paper to be published. We have dissected one paratype of D. bolivari (GZ50316) and two of D. tamaninii (ZMH838481 and ZMH838485) as well as 55 other females. The genital chamber is illustrated for D. bolivari paratype GZ50316 ( Fig. 1J View Fig ) and for D. tamaninii paratype ZMH838481 ( Fig. 1K View Fig ). The female genitalia of these two putative taxa are very homogeneous and of a single type. This is particularly true for the two paratypes (compare Figs 1J View Fig to 1K). This genital chamber is characterised by a cordiform shape, sometimes simply rounded, but above all by the margin of the sac, which is folded into a very characteristic accordion shape. The two oviducts are swollen at the base, well separated and generally directed downwards. This type of vaginal sac is very similar to those of D. tumidifrons Ribes, 1997 and D. escalerae , the closest relatives, but allows to separate these taxa from most other species in the subgenus Dicyphus (study in press). Again, this character cannot be used to separate D. bolivari and D. tamaninii .

Differential diagnosis. Dicyphus bolivari has the left paramere with a long apophysis and an endosoma with several small sclerites. It shares these characters with two other species, D. rubicundus and D. lindbergi ( Fig. 2 View Fig ). Dicyphus rubicundus can be easily separated from other species by the shape of its left paramere where the apex of the apophysis is continuous with the shaft without a clear demarcation unlike D. bolivari ( Fig. 2I View Fig compared to Figs 2C and 2F View Fig ). It can also be differenciated by its barcode sequence. The vaginal sac of the D. bolivari female is very similar to that of D. rubicundus . The case of D. lindbergi is less obvious. SANCHEZ & CASSIS (2018) commentated on D. lindbergi : ‘ Dicyphus lindbergi and D. bolivari are very similar in shape and colour (fig. 33), and the former species can only be confidently separated by the apophysis of the left paramere being shorter (figs 9C–F, 10E) and the fewer small endosomal lobal sclerites (N = 5 cf. N = 6–12) (figs 12C, 14C, D)’. We saw previously that these characters (size of the apophysis of the left paramere and the number of endosoma sclerites) were not relevant to differentiate D. bolivari from D. tamaninii because of their rather high variability. We were only able to examine two males of D. lindbergi but unfortunately no females. Furthermore, neither COI nor Cytb have been sequenced in this species to date. The male we have dissected from Cyprus ( Figs 2E–G View Fig ) has a 248 μm shaft of the apophysis and 7 endosomal sclerites which is in the range of D. bolivari . Therefore, the key provided by SANCHEZ & CASSIS (2018) does not allow us to differentiate this species from D. bolivari .

Molecular results. We produced 33 sequences of the COI standard barcode marker from specimens identified morphologicaly as D. bolivari or D. tamaninii from the following countries: England, France (Bretagne, Corse, Pays-de-la-Loire, Centre-Val-de-Loire, Grand-Est, Occitanie), Germany, Guernsey, the Netherlands, Spain and for the analysis we added the sequences published by SANCHEZ & CASSIS (2018) under the name D. bolivari from Spain and the Canary Islands. All specimens and their sequences are listed in Table 1.

SANCHEZ et al. (2006) produced a Cytb fraction of 381 bp from 5 specimens of Dicyphus identified as D. tamaninii . We constructed a phylogenetic tree ( Fig. 5 View Fig ) using their sequences and 20 sequences from our dataset ( Table 1). The Cytb sequences obtained from our specimens are exactly the same as those identified as D. tamaninii by SANCHEZ et al. (2006). The COI sequences of our 20 specimens are the same as those identified as D. bolivari by SANCHEZ & CASSIS (2018).

We also generated the complete mitochondrial genome for a specimen of D. bolivari / tamaninii (NCBI accession number: PP746700). The sequences identified as D. bolivari for COI by SANCHEZ & CASSIS (2018) and D. tamaninii for Cytb by SANCHEZ et al. (2006) are associated in the mitochondria of a single specimen.

Host plants. Dicyphus bolivari and D. tamaninii as defined by previous authors have a wide range of host plants, especially Solanaceae andAsteraceae, and at least two common host plant genera, i.e. Hyoscyamus and Solanum . For a list of the known host plant species see Table 4.

Distribution. Europe: Belgium ( AUKEMA 2020), Bosnia and Hercegovina ( PROTIć 1998), Channel Islands: Guernsey (new record), Croatia ( SANCHEZ & CASSIS 2018),

France including Corsica ( WAGNER & WEBER 1964), Germany ( SIMON 2020), Great Britain ( TELFER 2015), Greece ( SANCHEZ & CASSIS 2018), including Crete ( HEISS et al. 1993), Italy ( WAGNER 1951) including Sardinia (new record), Luxembourg ( AUKEMA 2020), Malta ( CARAPEZZA & MIFSUD 2015), Monaco (new record), the Netherlands ( AUKEMA 2020), Spain ( LINDBERG 1934). Asia: Cyprus (new record), Iran (ABD- RABOU & GHAHARI 2006), Israel ( LINNAVUORI 1961), Lebanon (MATOCQ & AZARD 2023), Asian part of Turkey (new record). North Africa: Canary Islands ( WAGNER 1951), Morocco ( WAGNER 1951, as D. maroccanus ), Tunisia ( WAGNER 1951).