Corresponding author: Vladimir A. Lukhtanov (lukhtanov@mail.ru)
Academic editor: V. Kuznetsova
Specimens with intermediate morphology are often considered to be the result of ongoing interspecific hybridization; however, this conclusion is difficult to prove without analysis of chromosomal and/or molecular markers. In the butterfly genus
Lukhtanov VA (2017) A new species of
Butterflies of the genus
Recent progress in improving our knowledge of relationships in
One of the most serious problems of the
While analyzing specimens of the genus
In an effort to analyze the origin of these unusual Israeli specimens and to determine their taxonomic status, their karyotype and morphology were studied and compared to those of
Specimens examined are deposited in the Zoological Institute of the Russian Academy of Sciences, St. Petersburg, Russia and in the McGuire Center for
Localities from where specimens of the
For genitalia preparation, abdomens removed from adults were soaked in hot (90°C) 10% KOH for 3–10 min. Then they were transferred to water, the genitalia were carefully extracted and examined under a stereo-microscope using a pair of preparation needles or a needle and a watchmaker’s tweezer. Once cleansed of all unwanted elements they were transferred and stored in tubes with glycerine. Cleansed genitalia armatures were handled, studied and photographed while immersed in glycerine, free from pressure due to mounting, and therefore free from the ensuing distortion. Genitalia photographs were taken with a Leica M205C binocular microscope equipped with a Leica DFC495 digital camera, and processed using the Leica Application Suite, version 4.5.0 software.
The terminology of genitalia structures follows
Butterfly photographs were taken with a Nikon D810 digital camera equipped with Nikon AF-S Micro Nikkor 105 mm lens.
Standard
88 samples were processed at the Canadian Centre for DNA Barcoding (
The barcode analysis involved 96
Sequences were aligned using the BioEdit software (
I used two criteria to evaluate the level of DNA barcode divergence between taxa and haplogroups. First, I calculated the number of fixed DNA substitutions, i.e. the number of invariable differences in the studied
Karyotypes were obtained from fresh adult males and processed as previously described (
(Fig.
26 males and 10 females collected on Mt. Hermon, Israel.
Four males with codes
Four males with codes
One male with codes
Ten males with codes
Two females with codes 25458 E06, BPALB149-16; 25458_E08 BPALB151-16. Israel, Mt. Hermon,
All paratypes are deposited in the Zoological Institute of the Russian Academy of Science (St. Petersburg).
(Fig.
Upperside: ground color orange-red; the wing markings are small and delicate when compared to those in
Underside: forewing ground color orange-red except for the apical part which is yellowish. Black markings delicate, reduced as compared with those of the upperside of the wing. Hindwing ground colour yellowish-white with two orange-red fascias. The red-orange submarginal fascia shows segmentation as the yellowish-white ground color spreads along the nervures. The orange-red macules of this fascia are bordered by back lunules from the outer side. From the inner side these macules are edged by black scales and additionally bordered by black lunules, giving the appearance that the proximal border of the submarginal fascia is doubly edged. Fringe white, checkered by black dots.
(Figs
The genus
Karyotype in male meiosis of
The same chromosome number (
Thus, no fixed karyotype difference is known to exist between
In
Male genitalia of
In
Lateral view of the inner face of right valva (left panel) and distal process (right panel). The angle between the valval dorsum and the distal process (left panel), and the keel of the distal process (right panel), are indicated by arrows.
In
Lateral view of the left side of aedeagus and (left panel), and lateral view of the right side of tegumen (right panel). The post-zonal dorso-lateral ridge (left panel) and lateral sclerotized element (right panel) are indicated by arrows.
The
The Bayesian tree of studied
The analysis recovered the
The first lineage (haplogroup P1) includes a huge range of
Fragment of the Bayesian tree of the studied
The haplogroup P1 includes also a female sample 17966_F12 possessing intermediate morphological characters between
The second lineage (haplogroup P2) is represented by three specimens from north Lebanon originally identified as
The third lineage (haplogroup P3) includes samples from NE Iran (
The forth lineage (haplogroup A), one of the most diverged lineages, is represented by samples from Mt. Hermon (
The fifth lineage (haplogroup H) includes samples of
Butterfly wing pattern and male genitalia morphology, as well as DNA barcodes certaintly indicate that
Points where
Three main vegetation belts have been described from Mt. Hermon: (i) evergreen Mediterranean maquis (300–1250 m); (ii) xero-montane open forest (1250–1850 m) and (iii) subalpine mountain steppe, or ‘‘Tragacanthic belt’’ (1850–2814 m) (
Habitat of
The name
Habitat of
Habitat of
To estimate the age of the haplogroup A (and the Israeli lineage as a whole) I used two calibration points: a lower rate of 1.5% uncorrected pairwise distance per million years estimated using a variety of invertebrates (
At the same time, the hypothesis that
Preimaginal stages of
The identity of the taxon described under the name
The same can be said about wing pattern. In fact, Belter was the first author who described two types of hindwing underside in
After Belter, the name
I should note that the identity of butterflies in these publications has never been clear, except for the monograph by Oorshot and Coutsis (2014) since at least three different groups of populations close to
The Gordian knot of this taxonomic and nomenclatural uncertainty was cut by Oorshot and Coutsis (2014) through a careful analysis of genitalia morphology, checking of all taxonomically important publications, studies of type material and subsequent designation of the lectotype of
The lectotype of
I should also note that, despite the valid lectotype designation resulting in this synonymy, the name
In this situation I see no other way than following the latest comprehensive revision (Oorshot and Coutsis 2014) that established the synonymy:
The
The identity and taxonomic status of the
I thank Dubi Benyamini, Vlad Dincă, Oleg Kosterin and Niklas Wahlberg who provided valuable comments on the paper. Asya Novikova (the Hebrew University of Jerusalem) and Elena Pazhenkova (Zoological Institute, St. Petersburg) helped with field studies in Israel. Elena Pazhenkova and Niklas Wahlberg donated their unpublished sequences: castaM8 and castaM9 (EP) and NW43-09 and NW43-10 (NW).The financial support for all molecular, chromosomal and morphological studies was provided by the grant N 14-14-00541 from the Russian Science Foundation to the Zoological Institute of the Russian Academy of Sciences. The travels to Israel and field studies of V. Lukhtanov were supported by RFBR grants 15-04-01581 and 15-29-02533. The work was partially performed using equipment of the ‘Chromas’ Core Facility, the Centre for Molecular and Cell Technologies and the Department of Entomology of St. Petersburg State University. A part of this equipment was purchased with support of the St. Petersburg University grant 1.40.490.2017.
List of
Taxon | Field ID | GenBank | Locality | Reference | |
---|---|---|---|---|---|
|
BPAL2759-15 |
|
Israel Hermon | This study | |
|
BPAL2236-13 |
|
Israel Hermon |
|
|
|
BPAL2235-13 |
|
Israel Hermon |
|
|
|
BPAL2234-13 |
|
Israel Hermon | This study | |
|
|
|
|
|
|
|
BPALB127-16 |
|
Israel Hermon | This study | |
|
BPALB126-16 |
|
Israel Hermon | This study | |
|
BPALB129-16 |
|
Israel Hermon | This study | |
|
BPALB133-16 |
|
Israel Hermon | This study | |
|
BPALB131-16 |
|
Israel Hermon | This study | |
|
BPALB125-16 |
|
Israel Hermon | This study | |
|
BPALB130-16 |
|
Israel Hermon | This study | |
|
BPALB149-16 |
|
Israel Hermon | This study | |
|
BPALB132-16 |
|
Israel Hermon | This study | |
|
BPAL3193-16 |
|
Israel Hermon | This study | |
|
BPAL3194-16 |
|
Israel Hermon | This study | |
|
BPAL3257-16 |
|
Israel Hermon | This study | |
|
BPAL3361-16 |
|
Israel Hermon | This study | |
|
BPAL3360-16 |
|
Israel Hermon | This study | |
|
BPAL3359-16 |
|
Israel Hermon | This study | |
|
BPALB138-16 |
|
Israel Hermon | This study | |
|
BPAL3192-16 |
|
Israel Hermon | This study | |
|
BPALB128-16 |
|
Israel Hermon | This study | |
|
BPALB151-16 |
|
Israel Hermon | This study | |
|
BPAL3191-16 |
|
Israel Hermon | This study | |
|
BPAL164-10 | RPVL-00164 |
|
Iran Fars | This study |
|
BPAL2302-14 |
|
Iran Fars | This study | |
|
BPAL2303-14 |
|
Iran Fars | This study | |
|
BPAL163-10 | RPVL-00163 |
|
Iran Fars | This study |
|
BPAL2307-14 |
|
Iran Lorestan | This study | |
|
BPAL2306-14 |
|
Iran Lorestan | This study | |
|
BPAL162-10 | RPVL-00162 |
|
Iran Zarde-Kuh Mts | This study |
|
BPAL2304-14 |
|
Iran Esfahan | This study | |
|
BPAL2305-14 |
|
Iran Esfahan | This study | |
|
NW85-3 |
|
Iran Fars Kuh-e Sorkh | This study | |
|
M8 |
|
Iran, Fereidunshahr | This study | |
|
M9 |
|
Iran, Fereidunshahr | This study | |
|
NW140-12 |
|
Iran Hamadan Alvand |
|
|
|
BPAL2354-14 |
|
Syria W Damascus |
|
|
|
BPAL2353-14 |
|
Syria W Damascus |
|
|
|
BPALB134-16 |
|
Israel Hermon 1800m |
|
|
|
BPALB150-16 |
|
Israel Hermon 1800m |
|
|
|
BPAL2704-14 |
|
Israel Hermon 2000m | This study | |
|
BPAL2686-14 |
|
Israel Zafririm 300m | This study | |
|
BPAL2857-15 |
|
Jordan | This study | |
|
BPAL2886-15 |
|
Israel Kfar-Adumim | This study | |
|
BPAL2885-15 |
|
Israel Kfar-Adumim | This study | |
|
BPAL2860-15 |
|
Jordan | This study | |
|
BPAL2898-15 |
|
Jsrael Kfar-Adumim | This study | |
|
BPAL3124-15 |
|
Israel Jerusalem |
|
|
|
BPAL2586-14 |
|
Israel Zafririm 300m | This study | |
|
BPAL2585-14 |
|
Israel Zafririm | This study | |
|
BPALB143-16 |
|
Israel Hermon 1730m |
|
|
|
BPALB142-16 |
|
Israel Hermon 1730m |
|
|
|
BPALB139-16 |
|
Israel Hermon 1730m |
|
|
|
BPALB137-16 |
|
Israel Hermon 1730m |
|
|
|
BPALB135-16 |
|
Israel Hermon 1730m |
|
|
|
BPALB136-16 |
|
Hermon 1730m |
|
|
|
BPAL2718-14 |
|
Israel Hermon 1650m |
|
|
|
BPAL2482-14 |
|
Afghanistan | This study | |
|
BPAL2469-14 |
|
Afghanistan | This study | |
NW34-10 |
|
Lebanon Bsharree |
|
||
NW43-09 |
|
Lebanon Laglong Mohafazat Jbail | This study | ||
NW43-10 |
|
Lebanon Laglong Mohafazat Jbail | This study | ||
|
BPAL2352-14 |
|
Iran Khorasan 2400m | This study | |
|
BPAL2351-14 |
|
Iran Khorasan 2400m | This study | |
|
BPAL2948-15 |
|
Iran Shahrud 2100m | This study | |
|
BPAL2959-15 |
|
Iran Shahrud 2100m | This study | |
|
BPAL2480-14 |
|
Iran Fars 2200m | This study | |
|
BPAL2481-14 |
|
Iran Fars 2200m | This study | |
|
BPAL2293-14 |
|
Iran Fars 2200m | This study | |
|
BPAL2295-14 |
|
Iran Esfahan 2000m | This study | |
|
BPAL2294-14 |
|
Iran Chahar Mahal-e Bakhtiari 2000m | This study | |
|
BPAL2982-15 |
|
Russia Daghestan 125m | This study | |
|
BPAL1704-12 |
|
Russia Daghestan 1750m | This study | |
|
BPAL2983-15 |
|
Russia Daghestan 125m | This study | |
|
BPAL1696-12 |
|
Russia Daghestan 2250m | This study | |
|
BPAL2977-15 |
|
Azerbaijan Talysh 1650m | This study | |
|
BPAL2984-15 |
|
Azerbaijan Talysh 1990m | This study | |
|
BPAL1689-12 |
|
Azerbaijan 600m | This study | |
|
BPAL2975-15 |
|
Azerbaijan Talysh 1650m | This study | |
|
BPAL2976-15 |
|
Azerbaijan Talysh 1650m | This study | |
|
BPAL2980-15 |
|
Azerbaijan Altyagach 1100m | This study | |
|
BPAL2979-15 |
|
Azerbaijan Altyagach 1100m | This study | |
|
BPAL2349-14 |
|
Iran Tehran 2000m | This study | |
|
NW120-11 |
|
Iran Ardabil 2600–2800 m |
|
|
BPAL2542-14 |
|
Armenia Megri 1800m | This study | ||
|
BPAL3098-15 |
|
Israel Hermon 2000m | This study | |
|
BPAL3125-15 |
|
Israel Jerusalem | This study | |
|
BPAL3122-15 |
|
Israel Jerusalem | This study | |
|
BPAL3121-15 |
|
Israel Jerusalem | This study | |
|
BPAL3116-15 |
|
Israel Jerusalem | This study | |
|
BPAL3115-15 |
|
Israel Jerusalem | This study | |
|
BPAL2858-15 |
|
Jordan | This study | |
|
BPAL2859-15 |
|
Jordan | This study | |
|
BPALB112-16 |
|
Israel Hermon 1450m | This study |