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ZOOTAXA

ISSN 1175-5326 (print edition)

ISSN 1175-5334 (online edition)Copyright © 2013 Magnolia Press

Zootaxa 3737 (2): 167–183

www.mapress.com/zootaxa/Article

http://dx.doi.org/10.11646/zootaxa.3737.2.5

http://zoobank.org/urn:lsid:zoobank.org:pub:A831D153-B117-4A22-AA4C-EDBAEE1FC31F

Systematical studies on the species of the subgenus Bombus (Thoracobombus)

(Hymenoptera: Apidae, Bombus Latreille) in Turkey

N.P. BARKAN* & A.M. AYTEKİNHacettepe University Faculty of Science Department of Biology Morphometry Laboratory Beytepe Campus 06800 Ankara, Turkey

*Corresponding Author. E-mail: pinarbarkan@gmail.com

Abstract

The aim of this study was to analyze the 12 species which belong to the subgenus Bombus (Thoracobombus) by identifying, collating and testing the applicability of geometrics morphometrics for distinguishing the species. This was carried out on 133 females and 42 males which were collected from various localities in Turkey. After digitizing landmarks on the right fore wings, 2-dimensional Cartesian coordinates were calculated and by Procrustes analysis the coordinates were standardized and

superimposed. Principal Components Analysis (PCA) and Canonical Variates Analysis (CVA) were performed to show the distribution of all species. Then, deformations which appeared in thin-plate splines were observed. Mean values of all the specimens were calculated and Sequential, Agglomerative, Hierarchical, and Nesting clustering method (SAHN) was performed with these data to obtain the dissimilarity trees. It can be concluded that all species were found to have consistently

different wing shapes from each other. In females, the species B. armeniacus, B. mesomelas and B. pomorum which resemble each other, were also found to be similar based on their wing morphometry. Both in females and males, the subspecies B.sylvarum citrinofasciatus and B. sylvarum daghestanicus and the species B. humilis and B. laesus exposed high similarity in wing morphometry. In males, results showed that the species B. armeniacus and B. mesomelas and the species B. humilis and B.zonatus have very similar wing shape.

Keywords: Bumble bees, Systematics, Geometric morphometrics, Principal Components Analysis, Canonical Variates Analysis

Introduction

Bumble bees have always been of interest to biologists’ as they are large, colorful and can be found in temperate regions where biologists are mostly active (Williams 1994). They are widely distributed especially in alpine, subalpine, temperate and arctic zones of the Palaearctic, Nearctic, Oriental and Neotropic regions of the world (Williams 1994, Cameron and Williams 2003, Hines 2008). Bumble bees are considered efficient pollinators as they are active foragers even at low temperatures where honey bee activity is limited (Free, 1993). As they have a crucial role in the pollination of many native and cultivated plant species, bumble bees are being commercially produced and are used in greenhouses (Free 1993, Hines et al. 2006).

It is estimated that there are over 250 species of Bombus (Williams et al. 2008) and with 47 species, Turkey is considered to be one of the countries with the highest species richness in the West-Palaearctic region (Aytekin et al. 2007, http://zoologie.umh.ac.be/hymenoptera/page.asp?id=103). In order to ease the relationships between the species of bumble bees in Bombus, a commonly used system based entirely on morphology was adopted a century ago (e.g., Dalla Torre 1880, Radoszkowski 1884, Robertson 1903, Vogt 1911, Skorikov 1914, 1923, Krüger 1920, Richards 1968). According to this classification, Bombus (Thoracobombus) was considered a subgenus of the genus Megabombus (Aytekin 2002). However, this subgeneric system of bumble bees was found to be rather complicated. This has generated a need to simplify the previously adopted system. According to a newly proposed subgeneric classification, it is suggested that B. (Thoracobombus) should be studied as one of the 15 main subgenera. As a consequence of this reduction, B. (Tricornibombus), B. (Exilobombus), B. (Fervidobombus), B.

Accepted by C. Rasmussen: 31 Oct. 2013; published: 19 Nov. 2013 167

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(Rhodobombus), B. (Laesobombus), B. (Eversmannibombus), B. (Mucidobombus) are included in B.(Thoracobombus) (Williams et al. 2008). Thus, by the addition of B. laesus from B. (Laesobombus), B. persicus

from B. (Eversmannibombus) and B. armeniacus and B. pomorum from B. (Rhodobombus), together with B.pascuorum, B. ruderarius, B. humilis, B. mlokosievitzii, B. sylvarum, B. zonatus, B. laesus, B. persicus, B.armeniacus, B. mesomelas, B. pomorum, B. velox, B. muscorum and B. brodmanni, B. (Thoracobombus) is characterized by 14 species in the West-Palaearctic.

Traditionally, bumble bees were identified only by morphology and especially the male genitalia was and is still very important for taxonomists (Dalla Torre 1880, Radoszkowski 1884, Robertson 1903, Vogt 1911, Skorikov 1914, 1923, Krüger 1920, Pittioni, 1937, 1938, 1939, Richards 1968, Reinig 1967, 1968, 1971, 1973, 1974, Alford 1975, Özbek 1983, Rasmont 1983, Ito 1985, Pawlikowski 1996, 1999, 2001, Aytekin and Çağatay 2003). They were also identified phylogenetically (Williams 1985, Williams 1994, Cameron & Williams 2003, Hines et al. 2006, Cameron et al. 2007) and by using classical and geometric morphometrics techniques (Ito 1987; Aytekin 2002, Aytekin and Çağatay 2003, De Meulemeester et al. 2009, Wappler et al. 2012). Later on, some other methods were engaged such as biochemical techniques (Pekkarinen and Teräs 1977, Pamilo et al. 1978, Pekkarinen 1979, Pekkarinen et al. 1979, Pekkarinen and Teräs 1993, Pamilo et al. 1987, Pamilo et al. 1997, Aytekin and Çağatay 2004) and molecular techniques which were conducted to enlighten the phylogeny of Bombus (Koulianos and Schmid-Hempel 2000, Cameron and Williams 2003, Kawakita et al. 2003, 2004, Hines et al. 2006, Cameron et al. 2007). Lately, very useful sexual pheromone analyses are being carried out on males (Urbanová et al. 2002, Terzo et al. 2003, Rasmont et al. 2005, Terzo et al. 2007). In Bombus, color patterns may show similarity between different species or can be highly variable within a single species. This makes classifications based on hair color unreliable (Terzo et al. 2007, Williams 2007). To overcome the difficulties in identification and in separating species, it was necessary to use alternative methods. Geometric morphometrics is one of them.

This very same problem appears in B. (Thoracobombus) which is a large subgenus represented by 14 species in Turkey. There are several unsolved problems in the taxonomy of B. (Thoracobombus) with respect to species status (Rasmont 1983, Aytekin and Çağatay 2003, Aytekin and Çağatay 2004). Geometric morphometrics may play a crucial role in determining the problems within the subgenera. For studies concerning bumble bees, geometric morphometrics is considered not only as an economical tool, but also an advantageous one as it gives results quickly and it is applicable even on old museum materials (Aytekin et al. 2007). Geometric morphometrics techniques have been conducted on Bombus impatiens by Klingenberg et al. (2001) as a tool for allometric studies but the first taxonomical attempt was driven by Aytekin et al. (2007) on the subgenus B. (Sibiricobombus) Vogt (1911). This technique was also used by De Meulemeester et al. (2009) on Bombus (Bombus) and by Wappler et al. (2012) on fossil bumble bee species so far. The aim of this study is to analyze the 12 species in B. (Thoracobombus) by identifying, collating and testing the applicability of geometric morphometrics methods on them.

Material and methods

The material consisted of bumble bee specimens from around the following cities in Turkey: Ankara, Eskişehir, Kırşehir, Nevşehir, Kayseri, Afyonkarahisar, Burdur, Isparta, Antalya, Karaman, Adana, Mersin, Niğde, Erzurum, Erzincan, Mersin, Trabzon, Artvin, Kastamonu, Rize, Giresun, Kars and Van. The specimens were collected between 1995 and 2009 and are kept in the Biology Department, Morphometry Laboratory of Hacettepe University, Turkey.

Material examined:

Bombus (Thoracobombus) pascuorum (Scopoli, 1763): 08-VIII-2001 Çatum creek, 2 km to Andon/ Rize (380 m) Prunella vulgaris L., 1 ♀ Leg. M. Aytekin; 08-08-2001 Güneyce-Trabzon Road/ Rize (140m) Trifolium

pratense L., 1 ♀ Leg. M. Aytekin; 07-08-2001 7 km to Uzungöl/ Trabzon (410 m) Trifolium pratense L., 4 ♀ Leg. M. Aytekin; 09-08-2001 Anzer Plateau/ Rize (1510 m) Trifolium pratense L., 1 ♂ Leg. M. Aytekin; 16-08-2001 Korucular Village-Murgul/ Artvin (727 m) Trifolium pratense L., 1 ♀ Leg. M. Aytekin; 17-08-2001 Camili Plateau/ Artvin (1672 m) Prunella vulgaris L. 1 ♀ Leg. M. Aytekin; 10-08-2005 Mısırlı Village/ Artvin 2 ♀ Leg. M. Aytekin; 16-08-2007 Zigana Pass, Limni Lake/ Giresun (2080 m) Cirsium echinus (Bieb.) Hand Mazz., 3 ♀

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Leg. F.Dikmen; 04-08-2005 Çoruh River proximity/ Artvin-Şavşat 2 ♀; 07-08-2005 Kafkasör Plateau/ Artvin 1 ♀Leg. F. Dikmen; 13-08-2005 Ayder Plateau/ Rize 2 ♀; 14-08-2005 Fırtına creek/ Rize 1 ♀ Leg. F. Dikmen; 16-08-2001 Korucular Village- Murgul/ Artvin (850 m) Cirsium osseticum (Adams) Petrak 1 ♀ Leg. M. Aytekin; 27-07-2004 Soğuksu National Park-Kızılcahamam/ Ankara 3 ♂ Leg. M. Aytekin; 03-08-2004 Soğuksu National Park-Kızılcahamam/ Ankara 1 ♂ Leg. F. Dikmen; 16-08-2007 Eğrisu Plateau/ Artvin Centaurea sp. 1 ♂ Leg. F. Dikmen; 12-08-2005 Oce Village-Ardeşen/ Rize 1 ♂ Leg. F. Dikmen.

Bombus (Thoracobombus) zonatus (Smith, 1854): 16-08-2006, Şuhut- Uzunpınar Village / Afyonkarahisar (1250 m), 3 ♀, 3 ♂ Leg. Y. Güler; 05-08-1995 Field Crops Central Research Institute, Haymana/ Ankara (1000 m) Consolida orientalis, 1 ♀ Leg. M. Aytekin; 08-08-1995 Beytepe Campus, Yenimahalle/ Ankara (1000m) 1 ♂ Leg. M. Aytekin; 23-07-1995 Bala-Ankara Road/ Ankara (1295 m) Ononis spinosa, 1 ♀ Leg. M. Aytekin; 23-08-1999 Beytepe Campus, Yenimahalle/ Ankara (980 m) 1 ♀, 5 ♂ Leg. M. Aytekin; 24-07-1997 Hacıbektaş-Gülşehir Road/ Nevşehir (1290 m) Morrubium parviflorum oligodon, 1 ♀ Leg. M. Aytekin; 24-07-1997 Hacıbektaş-Gülşehir Road/ Nevşehir (1250 m) Anchusa leptophylla, 1 ♀ Leg. M. Aytekin; 20-07-1997 Beypazarı/ Ankara, 2 ♀ Leg. M. Aytekin; 24-07-1997 Avanos- Kayseri Road, Mahmatlar/ Kayseri (1110 m) Salvia virgata, 2 ♀ Leg. M. Aytekin; 25-07-1997 Kırşehir-Aksaray road, Kesikköprü/ Kırşehir (800 m) Centaurea solstitialis, 2 ♀ Leg. M. Aytekin; 03-08-1997 Polatlı-Mihalıcçık Road, İğdecik/ Eskişehir (1100 m) Echium italicum, 1 ♀ Leg. M. Aytekin; 27-08-1999 Yukarıçavundur-Avcıova Road, Çubuk/ Ankara (1300 m) 1 ♂ Leg. M. Aytekin; 02-07-2009 5 km to Pınarlar Village, Tufanbeyli/ Adana (1451 m) Echium italicum L., 2 ♀ Leg. N. P. Barkan; 05-08-2009 Karabayır Plateau, Korkuteli-Finike Road/ Antalya.( 1691 m) 1 ♀ Leg. N.P. Barkan; 17-07-2009 Avcıpınar-Yeniköy Road/ Adana (1549 m) 2 ♀ Leg. N. P. Barkan.

Bombus (Thoracobombus) mlokosievitzii Rad., 1877: 10-08-2005 Mısırlı Village/ Artvin 3 ♀.Bombus (Thoracobombus) sylvarum (L., 1761): 23-07-1995 Elmadağ-Bala Road, Elmadağ/ Ankara (940 m)

Ononis spinosa, 4 ♀ Leg. M. Aytekin; 22-07-1995 Yukarıçanlı-Sipahiler Crossroad, Kızılcahamam/ Ankara (1370 m) Salvia virgata, 1 ♀ Leg. M. Aytekin; 22-08-1995 Yukarıçanlı-Sipahiler Crossroad, Kızılcahamam/ Ankara (1370 m) Cousinia spp., 2 ♀ Leg. M. Aytekin; 12-08-1995 Elmadağ-Bala Road, Elmadağ/ Ankara (1295 m) Helianthus annuus, 1 ♀ Leg. M. Aytekin; 12-08-1995 Elmadağ-Bala Road, Elmadağ/ Ankara (1000 m) Echium

italicum, 1 ♀ Leg. M. Aytekin; 25-08-2007 Sultandağı- Yeşilçiftlik/ Afyonkarahisar 1 ♀ Leg. Y. Güler; 02-09-1999 Işık Mountain-Kızılcahamam/ Ankara (2000 m) Cousinia sp., 1 ♂ Leg. M. Aytekin ; 20-08-1999 Beynam Atatürk Forest Road, Bala/ Ankara (1191 m) 1 ♂ Leg. M. Aytekin; 13- 08-2001 Ayder Plateau National Park/ Rize Prunella

vulgaris L. (1800 m) 1 ♀ M. Aytekin; 11-08-2009 4km to Dokuzdolananlar Pass/ Kayseri Onopordum

polycephalum Boiss. 5 ♀ Leg. N. P. Barkan; 18-08-2009 15 km to Çamardı/ Niğde Onopordum sibthorpianum

Boiss. and Heldr. 2 ♀ Leg. N. P. Barkan; 18-08-2009 15 km to Çamardı/ Niğde Alkanna orientalis (L.) Boiss. var.orientalis 1 ♀ Leg. N. P. Barkan; 16-08-2007 Zigana Pass, Limni Lake/ Giresun (2080 m.) Cirsium echinus Hand.-Mazz 1 ♀ Leg. F. Dikmen; 11-08-2009 4km to Dokuzdolananlar Pass / Kayseri Onopordum polycephalum Boiss. 3 ♂ Leg. N. P. Barkan; 18-08-2009 15 km to Çamardı/ Niğde Onopordum sibthorpianum Boiss. and Heldr. 1 ♂ Leg. N. P. Barkan; 25-08-2008 Nemrut Crater Lake/ Van (2262 m) 1 ♂ Leg. Ç. Özenirler.

Bombus (Thoracobombus) humilis Illiger, 1806: 03-08-2001, Kızılcahamam/ Ankara 1 ♀ Leg. Y. Güler; 26-06-2008, Pınarbaşı/ Kastamonu, 2 ♀ Leg. F. Dikmen; 07-06-2006, Şuhut-Başören Village/ Afyankarahisar (1624 m) 1 ♀ Leg. M. Karataş; 20-08-2008, Kolçekmezdağı Pass/ Erzincan (2140 m) 2 ♂ Leg. F. Dikmen.

Bombus (Thoracobombus) ruderarius (Fab., 1843): 08-08-2009 Alidağ- Talaş/ Kayseri (1862 m) Anchusa

sp., 1 ♀ Leg. N. P. Barkan; 09-08-2009 Aksu Plateau/ Kayseri (1890 m) Anchusa sp. 4 ♀ Leg. N. P. Barkan; 04-08-2009 Söbüce Village Plateau/ Burdur-Antalya (1840 m) Onopordum sp. 6 ♀ Leg. N. P. Barkan; 08-08-2009 Alidağ-Talaş/ Kayseri (1862 m) 2 ♂ Leg. N. P. Barkan; 09-08-2009 Aksu Plateau Road/ Kayseri (1890 m) 2 ♂ Leg. N. P. Barkan; 11-08-2009 Söbüce Village Plateau/ Burdur-Antalya Onopordum sibthorpianum Boiss. and Heldr. 6 ♀ Leg. N. P. Barkan; 5-07-2006 3 km after Kızılören-Soğucak Village/ Afyonkarahisar (1578 m) 2 ♀ Leg. Y. Güler.

Bombus (Thoracobombus) pomorum (Panzer, 1805): 04-08-2009, Gönen Mountain Road/ Isparta, 1 ♀ Leg. N. P. Barkan; 19-08-2009, Üçkapılı Village/ Niğde, Onopordum sp., 2 ♀ Leg. N. P. Barkan.

Bombus (Thoracobombus) mesomelas (Gersteacker, 1869): 02-09-1999 Işık Mountain- Kızılcahamam/ Ankara Cousinia sp. (2000 m) 8 ♂ Leg. M. Aytekin; 27-08-1999 Yukarıçavundur-Avcıova Road, Çubuk/ Ankara (1300 m) 1 ♂ Leg. M. Aytekin; 20-08-1999 Beynam Atatürk Forest Road- Bala/ Ankara (1191 m) 1 ♂ Leg. M. Aytekin; 20-08-2008 Kolçekmez Mountain Pass/ Erzincan (2140 m) 1 ♂ Leg. F. Dikmen; 04-08-2009 Söbüce

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Village Plateau/ Antalya (1840 m) Onopordum sibthorpianum Boiss. and Heldr., 1 ♂ Leg. N. P. Barkan; 17-08-2008 Ovacık Üç kilise, Rizekent proximity/ Erzurum, Centaurea sp., 2 ♀ Leg. F. Dikmen.

Bombus (Thoracobombus) armeniacus (Rad.), 1877 : 23-07-1995, Elmadağ-Bala Road, 6 km, north of Karacahasan, Elmadağ/ Ankara (1295 m) Ononis spinosa 1 ♀ Leg. M. Aytekin; 05-07-2006 Kızılören- Soğucak Village/ Afyonkarahisar (1540 m) 2 ♀ Leg. Y. Güler; 23-07-2009 5-6 km from Anamur-Ermenek Road (1833 m.) Phlomis armeniaca Wild., 1 ♀ Leg. N. P. Barkan; 21-07-2009 Çal location- Mut/ Mersin (1769 m) 1 ♀ Leg. N. P. Barkan; 17-08-2008 Ovacık üç kilise, Rizekent proximity/ Erzurum Centaurea sp. 2 ♀ Leg. F. Dikmen; 08-08-2009 Alidağ-Talaş/ Kayseri Anchusa leptophylla Roemer and Schultes subsp. leptophylla(1862 m.) 2 ♀ Leg. N. P. Barkan; 11-08-2009 4 km. to Dokuzdolananlar Pass/ Kayseri Onopordum polycephalum Boiss. (1705 m) 1 ♀ Leg. N. P. Barkan; 21-VII-2009 Şıh Fountain-Mut/ Mersin Onopordum sp. (1729 m) 1 ♀; 18-VIII-2009 Bolkar Mountain / Niğde Asteraceae, 2 ♀ Leg. F. Dikmen; 16-07-2008 Sandıklı-Çakmaktepe Pass/ Afyonkarahisar (1884 m) 1 ♀ Leg. Y. Güler; 05-07-2006 Kızılören-Soğucak Village/ Afyonkarahisar (1540 m) 1 ♀ Leg. Y. Güler; 04-08-2009 Söbüce Village Plateau/ Antalya Onopordum sp. (1840 m) 1 ♀ Leg. N. P. Barkan; 03-08-2003 Akşehir-Isparta Road Anhcusa sp. (1555 m) 1 ♀; 23-07-2009 27 km to Taşkent/ Karaman Boraginaceae (1867 m)1 ♀ Leg. N. P. Barkan; 23-07-2009 27 km to Taşkent/ Karaman Onopordum sp.(1867 m) 1 ♀ Leg. Ç. Özenirler; 17-07-2009 Avcıpınar- Yeniköy Road/ Adana (1549 m) Salvia virgata Jacq., 1 ♀ Leg. N. P. Barkan; 02-09-1999, Işık Mountain- Kızılcahamam/ Ankara (2000 m) Cousinia sp., 1 ♂.

Bombus (Thoracobombus) laesus (Morawitz, 1875): 17-VII- 2008 Çay-Çayıyazı Village/ Afyonkarahisar (1014 m) 1 ♀ Leg. N. P. Barkan; 04-06-2009 Şıh Fountain-Mut/ Mersin (1745 m) Alkanna orientalis (L.) Boiss. var. orientalis, 2 ♀ Leg. N. P. Barkan; 21-07-2009 Şıh Fountain- Mut/ Mersin (1729 m) Onopordum sibthorpianum

Boiss. and Heldr. 2 ♀ Leg. N. P. Barkan; 21-07-2009 Şıh Fountain- Mut/ Mersin (1729 m) 2 ♀ Leg. N. P. Barkan; 21-07-2009 Çal location- Mut/ Mersin (1881 m) 4 ♀ Leg. N. P. Barkan; 23-07-2009 5-6 km from Anamur-Ermenek (1833 m) 1 ♀ Leg. N. P. Barkan; 17-07-2009 Avcıpınar-Yeniköy Road/ Adana (1549 m) 2 ♀ Leg. N. P. Barkan; Çıldır Lake Road, Taşbaşı Village/ Kars (1982 m) Centaurea sp. 1 ♀ Leg. F. Dikmen.

Bombus (Thoracobombus) persicus Rad., 1881 : 22-06-2006 Şuhut-Kumalar Mountain/ Afyonkarahisar (2192 m) 1 ♀ Leg. Y. Güler; 02-07-2009 Tufanbeyli-Devli Road, Hanyeri Village/ Kayseri (1880 m) 1 ♀ Leg. N. P. Barkan; 11-08-2009 4km to Dokuzdolananlar Pass/ Kayseri (1705 m) Onopordum polycephalum Boiss. 2 ♀ Leg. N. P. Barkan; 17-08-2008 Ovacık üç kilise, Rizekent proximity/ Erzurum Centaurea sp. 1 ♀ Leg. F. Dikmen; 20-08-2008 Kolçekmezdağı Pass/ Erzincan (2140 m) 1 ♀ Leg. F. Dikmen.

Bombus (Thoracobombus) velox (Skorikov, 1914) : 15-08-2007 Çıldır Lake Road, Taşbaşı Köyü/ Kars (1982 m) Centaurea pterocaula Trautr., 3 ♀ Leg. F. Dikmen.

Geometric morphometrics analysis

This analysis was performed on 133 specimens of which 12 species were female and 42 specimens of which 7 species were male. While conducting PCA and/ or CVA, 2 subspecies of B. sylvarum, B. sylvarum citrinofasciatus

and B. sylvarum daghestanicus, were regarded as 2 separate groups. For this reason, 13 groups for the females and 8 groups for the males were adopted.

For geometric morphometrics analysis, the right fore wings of 175 specimens which belong to the species B. pascuorum, B. ruderarius, B. humilis, B. mlokosievitzii, B. sylvarum, B. zonatus, B. laesus, B. persicus, B. armeniacus, B. mesomelas, B. pomorum, B. velox were removed from their mesothorax using forceps and fixed onto slides using Entellane (Merck, KgaA, Darmstadt, Germany). No dye was applied to the wings because of clear venation. All slides were photographed using a Leica MZ-7.5 stereoscopic zoom dissection microscope integrated with a DC-300 digital camera system. After the digitization procedure, specific codes were given to the photographs which were then archived. The photographs were first organized and made compatible to the TPS series by using TPSUTIL 1.44 (Rohlf 2009). Secondly, 20 landmarks were digitized in accordance with Aytekin et al. (2007) (Figure 1) (TPSDIG 2.12; Rohlf 2008) and 2-dimensional Cartesian coordinates were obtained. Before starting the analysis, TPSSMALL 1.20 (Rohlf 2003) and TPSSUPER 1.14 (Rohlf 2004) were conducted. TPSSMALL 1.20 was used to determine whether the amount of variation in shape in a data set was small enough to permit statistical analyses to be performed in the linear tangent space approximate to Kendall's shape space that is non-linear. The average configuration is also often referred to as a "consensus" configuration and it serves as the "reference" configuration (that defines the point of tangency between shape space and the approximating tangent space) in the computation of the thin-plate splines. 2 options are provided to control how the specimens are scaled

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after they are aligned to the consensus configuration. The default is to make each specimen to have a centroid size of one. TPSSUPER 1.14 was used to compute the orthogonal least-squares Procrustes average configuration of landmarks using the generalized orthogonal least-squares procedures described in Rohlf and Slice (1990). It is also often referred to as a “consensus configuration and it serves as the “reference configuration” in the computation of the thin-plate splines.

FIGURE 1. Right forewing of a bumble bee wing with 20 landmarks.

The coordinates were then standardized by carrying out Generalized Procrustes Analysis (GPA) using Integrated Morphometrics Package (IMP) (Zelditch et al. 2007) CoordGen6f. By doing so, the differences between landmark configurations (scale, ordination and orientation) were eliminated. The data was then examined according to its relative warps in TPSRELW 1.45 (Rohlf 2007). The distribution of structural shape differences were shown on the first two principal components (PC) in PCAGen6p, IMP. In CVAGen6n, IMP canonical variates analysis was conducted and the distribution of individuals on the first two canonical variates (CV) was obtained. By conducting Multivariate Analysis of Variance (MANOVA) the statistical significance of the distribution was analyzed and detection was made to see if there were any differences between groups or not. The data which was standardized via GPA rotation in Morpheus (Slice 2007) was then examined using thin plate spline in the same program. The differences between every species’ consensus configurations from the general average reference were summarized using graphics. Thus, variation among groups was observed by determining the landmarks which caused deformation the most. The average values obtained from the same program were then used to construct a dissimilarity tree using SAHN in the Numerical Taxonomy and Multivariate Analysis System (NTSYS) 2.10 Software (Rohlf 2000).

Results

Females: 133 female specimens which belong to B. armeniacus, B. humilis, B. laesus, B. mesomelas, B. mlokosievitzii, B. pascuorum, B. persicus, B. pomorum, B. ruderarius, B. sylvarum, B. velox and B. zonatus were analyzed. After placing the landmarks in TPSDIG.12 (Rohlf 2008), the amount of variation in the data necessary to conduct thin plate spline analysis was tested in TPSSMALL 1.20 (Rohlf 2003). The amount of variation was found to be enough to show similarity among species.

Later on, the adequacy of the number of landmarks used was tested in TPSSUPER 1.14 (Rohlf 2004). After receiving favorable results from these 2 programs Procrustes was performed to determine the distribution of the landmarks and to detect if there was any deviation in distribution (Figure 2).

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FIGURE 2. Consensus configuration of the landmarks in females after Procrustes superimposition.

FIGURE 3. Distribution of female individuals along the first two principal components. Horizontal axis represents PC1,

vertical axis represents PC2 and each dot represents one individual.

PCA: The subspecies B. sylvarum citrinofasciatus and B. sylvarum daghestanicus were regarded as 2 separate groups for the grouping necessary to perform PCA and CVA. Thus, 13 groups were adopted. In PCAGen6p, IMP

(Zelditch et al. 2007) the distribution of individuals along the 1st and 2nd (PC1 and PC2), on the 2nd and 3rd (PC2 and

PC3) and on the 1st and 3rd (PC1 and PC3) principal components were examined (Figure 3–5). Relative warps: The relative warps were determined in TPSRELW (Rohlf 2007). According to the results, the

landmarks having the highest relative scores were found to be the 15th and 16th, while the landmark having the

lowest relative scores was found to be the 1st. Results concerning the value of variance showed that the values S²= 0.00011262, S²= 0.00011132 and S²= 0.00010971 referring to landmarks number 13, 17 and 19 respectively

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showed that these landmarks had the highest value of variance, while the value S²= 0.0002081 representing the landmark number 14 showed that this landmark had the lowest value of variance.

FIGURE 4. Distribution of female individuals along the 1st and 3rd principal components. Horizontal axis represents

PC1, vertical axis represents PC3 and each dot represents one individual.

FIGURE 5. Distribution of female individuals along the 2nd and 3rd principal components. Horizontal axis represents

PC2, vertical axis represents PC3 and each dot represents one individual.

Deformation grids: In the Morpheus program, the variation of the fore wings between the species was shown on deformation grids and thin plate spline analysis was conducted. The deformations were amplified 2-fold to observe the differences between species with ease. Compared with the others, the deformation grids of B. sylvarum

citrinofasciatus and B. sylvarum daghestanicus showed low variability. To indicate this, the deformation grid of only these subspecies is included in the context of the study and it is shown in the figure below (Figure 6).

CVA: CVAGen6p, IMP analysis was conducted to determine if there is any significant difference between the groups. MANOVA was conducted in the same program and according to the values obtained it was concluded that all 13 groups were significantly different from each other (Table 1).

CANOVAR was conducted in CVAGen6n, IMP and the distribution of individuals on the 1st two canonical variants was determined (Figure 7).

Dissimilarity tree: The mean values were obtained from the tps values by using the Morpheus software. These

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mean values were then grouped in NTYSYS using SAHN tree. A dissimilarity tree was then constructed using unweight pair group method with arithmetic mean (UPGMA) (Figure 8).

TABLE 1. MANOVA table of 13 groups for females.

FIGURE 6. Deformation grid of female individuals for B. sylvarum citrinofasciatus and B. sylvarum daghestanicus. The

deformation grid shows low variability between the subspecies B. sylvarum citrinofasciatus and B. sylvarum

daghestanicus (exaggeration was made twice for a better visualization).

Males: 42 male specimens which belong to B. armeniacus, B. humilis, B. mesomelas, B. pascuorum, B.ruderarius, B. sylvarum and B. zonatus were analyzed. After placing the landmarks in TPSDIG 2.12 (Rohlf 2008), the amount of variation in the data necessary to conduct thin plate spline analysis was tested in TPSSMALL 1.20 (Rohlf 2003). The amount of variation was found to be enough to show similarity among species.Later on, the adequacy of the number of landmarks that were used was tested in TPSSUPER 1.14 (Rohlf 2004). After receiving favorable results from these 2 programs Procrustes was performed to determine the distribution of the landmarks and to detect if there was any deviation in distribution (Figure 9).

PCA: The subspecies B. sylvarum citrinofasciatus and B. sylvarum daghestanicus were taken as 2 separate groups for the grouping necessary to perform PCA and CVA. Thus, eight groups were adopted.

In PCAGen6p, IMP (Zelditch et al. 2007) the distribution of individuals along the 1st and 2nd (PC1 and PC2), on

the 2nd and 3rd (PC2 and PC3) and on the 1st and 3rd (PC1 and PC3) principal components were examined (Figure 10-12).

Relative warps: The relative warps were determined in TPSRELW (Rohlf 2007). According to the results the landmarks having the highest relative scores were found to be number 15 and 16, while the landmark having the lowest relative scores was found to be number 1 similar to the results of the female species. Results concerning the value of variance showed that the values S²= 0.00012474 and S²= 0.00010392 referring to landmarks number 13 and 17 respectively showed that these landmarks had the highest value of variance, while the value S²= 0.0001913 representing the landmark number 7 showed that this landmark had the lowest value of variance.

Lambda chi2 P value

Axis 1 0.0000 1292.4837 p<2.22045e-016

Axis 2 0.0001 1032.8196 p<2.22045e-016

Axis 3 0.0004 831.7744 P<2.22045e-016

Axis 4 0.0023 653.1081 P<2.22045e-016

Axis 5 0.0117 477.7408 P=1.33227e-015

Axis 6 0.0348 360.8599 P=2.97709e-009

Axis 7 0.0873 262.0934 P=6.23927e-005

Axis 8 0.1872 180.1171 P=0.0253347

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FIGURE 7. The distribution of groups on the first two canonical variates. Horizontal axis represents CV1, vertical axis

represents CV2 and each dot represents one individual.

FIGURE 8. The UPGMA tree which shows the comparison of the 13 taxa.

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Deformation grids: In the Morpheus program, the variation of the fore wings between the species was shown on deformation grids and thin plate spline analysis was conducted. The deformations were amplified 2-fold to observe the differences between species with ease. As in females, the deformation grid of only B. sylvarum

citrinofasciatus and B. sylvarum daghestanicus are shown in the figure below (Figure 13).CVA: CVAGen6p, IMP analysis was conducted to decide whether or not there is a significant difference

between the groups. MANOVA was conducted in the same program and according to the values obtained it was concluded that all 8 groups were significantly different from each other (Table 2).

FIGURE 9. Consensus configuration of the landmarks in males after Procrustes superimposition.

FIGURE 10. Distribution of male individuals along the first two principal components. Horizontal axis represents PC1,

vertical axis represents PC2 and each dot represents one individual.

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CANOVAR was conducted in CVAGen6n, IMP and the distribution of individuals on the first two canonical variants was determined (Figure 14).

Dissimilarity tree: The mean values were obtained from the tps values by using the Morpheus software. These mean values were then grouped in NTYSYS using SAHN Clustering Method based on Euclidian distance. Then a dissimilarity tree was constructed using unweight pair group method with arithmetic mean (UPGMA) (Figure 15).

FIGURE 11. Distribution of male individuals along the 1st and 3rd principal components. Horizontal axis represents PC1,

vertical axis represents PC3 and each dot represents one individual.

FIGURE 12. Distribution of male individuals along the 2nd and 3rd principal components. Horizontal axis represents PC2,

vertical axis represents PC3 and each dot represents one individual.

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FIGURE 13. Deformation grid of male individuals for B. sylvarum citrinofasciatus and B. sylvarum daghestanicus. The

deformation grid shows low variability between the subspecies B. sylvarum citrinofasciatus and B. sylvarum

daghestanicus (exaggeration was made twice for a better visualization).

TABLE 2. MANOVA table of 8 groups for males.

FIGURE 14. The distribution of groups on the first two canonical variates. Horizontal axis represents CV1, vertical axis

represents CV2 and each dot represents one individual.

Lambda chi2 P value

Axis 1 0.0000 1114.1613 p<2.22045e-016

Axis 2 0.0000 413.8609 p<2.22045e-016

Axis 3 0.0000 251.8005 P=6.08378e-009

Axis 4 0.0002 163.0219 P=5.39671e-005

Axis 5 0.0100 89.7915 P=0.0184377

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FIGURE 15. The UPGMA tree which shows the comparison of the 8 taxa.

Discussion

In this study, geometric morphometrics techniques have been used to analyze the taxonomical relationships between 12 species of B. (Thoracobombus). By using landmark based geometric morphometrics, wing shapes were identified and the analysis of the differences and similarities among these species determined the systematical relationships within B. (Thoracobombus). Most of the findings were found to be compatible with those of Aytekin (2002). The fact that color patterns seen in the species of Bombus show similarity or can be highly variable within species is also seen in B. (Thoracobombus) which creates problems in the systematics of the subgenus (Rasmont 1983, Aytekin and Çağatay 2003, Aytekin and Çağatay 2004, Williams 2007). To overcome such difficulties, geometric morphometrics was adopted to solve problems within B. (Thoracobombus) in this study.

According to MANOVA results conducted in IMP, all groups were found to be distinct and different from each other showing that geometric morphometrics is applicable in identifying the species within B. (Thoracobombus). A closer look on the PCA results in both females and males showed that some species were clustered together and had similar wing shape morphology. According to the PCA results in females, B. armeniacus, B. mesomelas and B.pomorum were found to appear close to each other in distribution in PC1 and PC2; and so were B. sylvarum

citrinofasciatus, B. sylvarum daghestanicus, B. zonatus and B. laesus. Individuals from B. mesomelas were found to be close with those of B. mlokosievitzii in PC1 and PC3. In CVA, the following species were found to be similar to each other: B. humilis and B. laesus; B. armeniacus, B. mesomelas, B. pomorum and B. velox; B. sylvarum

citrinofasciatus and B. sylvarum daghestanicus. The UPGMA tree showed similar results with that of CVA. According to the PCA results in males, individuals which belong to the species B. mesomelas and B. armeniacus

were found to appear close to each other in distribution both in PC1 and PC2, and PC1 and PC3, which shows that they are similar in wing shape. Individuals, which belong to B. ruderarius, B. humilis, B. zonatus, B. sylvarum

citrinofasciatus and B. sylvarum daghestanicus, were intertwined with each other. In CVA, similar with the results

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in females, it was found out that B. sylvarum citrinofasciatus and B. sylvarum daghestanicus coincided with each other. In the UPGMA tree, B. armeniacus and B. mesomelas; B. humilis and B. zonatus; and finally B. sylvarum

citrinofasciatus and B. sylvarum daghestanicus were clustered together. B. humilis and B. zonatus had also formed a cluster in a previous study by Aytekin and Çağatay (2004) where electrophoretic analysis was performed. The species B. armeniacus, B. mesomelas and B. pomorum which resemble each other in showing similar color patterns, were also found to be similar based on their wing morphometry in females. The same cluster was seen in the Bayesian tree based on five different gene sequences (Cameron et al. 2007). The species B. humilis and B.laesus exposed similar wing morphometry. In males, the species B. armeniacus and B. mesomelas and the species B. humilis and B. zonatus showed similar wing morphometry. Both in females and males, the subspecies B.sylvarum citrinofasciatus and B. sylvarum daghestanicus also showed similar wing morphometry. But to make a decision on their status, a comprehensive study including all subspecies of this species is needed.

In summary, this study revealed promising results for the use of geometric morphometrics in the analyses of B. (Thoracobombus). All species formed distinct groups concluding that geometric morphometrics is capable of separating and identifying these species. According to the PCA and CVA graphs, species clustering together were mostly close species with similar morphology. Further studies which would integrate geometric morphometrics with molecular methods would help enlighten the relationships within the subgenus.

Acknowledgments

This study was part of the requirements for the MSc degree in Hacettepe University and was supported by Hacettepe University Research Foundation Project No: 0701601016 (10-10-2007/10-10-2010). We are grateful to Dr. Claus Rasmussen and Dr. Torsten Wappler for comments and suggestions that helped to improve this article. We would like to thank Dr. Pierre Rasmont for letting us examine his diverse bumble bee collection in Mons, Belgium and Dr. Fatih Dikmen, Çiğdem Özenirler and Kürşat Koyuncu for their great support during field trips. We would also like to thank Candan McSweeney for editing the English.

References

Alford, D.V. (1975) Bumblebees. Davis-Poynter, London, 352 pp.Aytekin, A.M. (2002) İç Anadolu Bölgesi Apidae (Hymenoptera) Familyası Üzerinde Sistematik Araştırmalar ile Sorunlu

Gruplarda Morfometrik ve Biyokimyasal Yöntemlerin Uygulanması. Hacettepe Üniversitesi Fen Bilimleri Enstitüsü, Ankara, 152 pp.

Aytekin, A.M. & Çağatay, N. (2003) Systematical Studies on Megabombus (Apidae: Hymenoptera) Species in Central Anatolia. Turkish Journal of Zoology, 27, 195–204.

Aytekin, A.M. & Çağatay, N. (2004) Allozyme Variability In Some Central Anatolian Bumble Bee (Bombus, Apidae, Hymenoptera) Species. Mellifera, 4 (8), 34–37.

Aytekin, A.M., Terzo, M., Rasmont, P. & Çağatay, N. (2007) Landmark based geometric morphometric analysis of wing shape in Sibiricobombus Vogt (Hymenoptera: Apidae: Bombus Latreille). Annales de la Société Entomologique de France (Nouvelle série), 43 (1), 95–102. http://dx.doi.org/10.1080/00379271.2007.10697499

Cameron, S.A. & Williams, P.H. (2003) Phylogeny of bumble bees in the New World subgenus Fervidobombus (Hymenoptera: Apidae): congruence of molecular and morphological data. Molecular Phylogenetics and Evolution, 28, 552–563. http://dx.doi.org/10.1016/S1055-7903(03)00056-3

Cameron, S.A., Hines, H.M. & Williams, P.H. (2007) A comprehensive phylogeny of the bumble bees (Bombus). Biological Journal of the Linnean Society, 91, 161–188. http://dx.doi.org/10.1111/j.1095-8312.2007.00784.x

Dalla Torre, K.W. (1880) Unsere Hummel- (Bombus) Arten. Der Naturhistoriker, 2, 40–41.De Meulemeester, T., Aytekin, A.M., Valterova, I. & Rasmont, P. (2009) Landmark based geometric morphometrics analysis of

wing shape as a tool for Bombus s.str. taxonomy (Hymenoptera: Apidae). 6th Symposium National de Morphometrie et Evolution des Formes, Montpellier, May 27–28, France, 49 pp.

Dryden, I.L. & Mardia, K.V. (1998) Statistical Shape Analysis. John Wiley and Sons, New York, 347 pp.Free, J.B. (1993) Insect Pollination of Crops, 2nd Edition. Academic Press, London, UK, 684 pp.Heinrich, B. (1979) Bumblebee Economics. Cambridge, MA, Harvard University Press, 245 pp.

BARKAN & AYTEKİN180 · Zootaxa 3737 (2) © 2013 Magnolia Press

TERMS OF USE This pdf is provided by Magnolia Press for private/research use. Commercial sale or deposition in a public library or website is prohibited.

Hines, H.M., Cameron, S.A. & Williams, P.H. (2006) Molecular phylogeny of the bumble bee subgenus Pyrobombus(Hymenoptera: Apidae: Bombus) with insights into gene utility for lower-level analysis. Invertebrate Systematics, 20, 289–303. http://dx.doi.org/10.1071/is05028

Hines, H.M. (2008) Historical biogeography, divergence times, and diversification patterns of bumble bees (Hymenoptera: Apidae: Bombus). Systematic Biology, 57, 58–75.

Ito, M. (1985) Supraspesific Classification of Bumblebees Based on Characters of Male Genitalia. Zoological Section. The Institute of Low Temperature Science Publ. Hokkaido University, Japan, 143 pp.

Ito, M. (1987) Geographic Variation of an East Asian Bumblebee Bombus diversus in some Morphometric Characters (Hymenoptera, Apidae). Kontyu, Tokyo, 55 (2), 188–201.

Kawakita, A., Sota, T., Ascher, J.S., Ito, M., Tanaka, H. & Kato, M. (2003) Evolution and phylogenetic utility of alignment gaps within intron sequences of three nuclear genes in bumble bees (Bombus). Molecular Biology and Evolution, 20, 87–92. http://dx.doi.org/10.1093/molbev/msg007

Kawakita, A., Sota, T., Ito, M., Ascher, J.S., Tanaka, H., Kato, M. & Roubik, D.W. (2004) Phylogeny, historical biogeography, and character evolution in bumble bees (Bombus: Apidae) based on simultaneous analysis of three nuclear gene sequences. Molecular Phylogenetics and Evolution, 31, 799–804. http://dx.doi.org/10.1016/j.ympev.2004.04.001

Klingenberg, C.P., Badyaev, A.V., Sowry, S.M. & Beckwith, N.J. (2001) Inferring developmental modularity from morphological integration: analysis of individual variation and asymmetry in bumblebee wings. American Naturalists, 157, 11–23. http://dx.doi.org/10.1086/317002

Koulianos, S. & Schmid-Hempel, P. (2000) Phylogenetic relationships among bumblebees (Bombus, Latreille) inferred from mitochondrial Cytochrome b and Cytochrome Oxidase I sequences. Molecular Phylogenetics and Evolution, 14, 335–341. http://dx.doi.org/10.1006/mpev.1999.0621

Krüger, E. (1920) Beiträge zur Systematik und Morphologie der mittel-europäischen Hummeln. Zoologische Jahrbücher (Systematik, Ökologie und Geographie der Tiere), 42, 289–464.

Özbek, H. (1983) Doğu Anadolu’nun bazı yörelerinde Bombinae (Hym.: Apoidea, Bombidae) türleri üzerinde taksonomik ve bazı biyolojik çalışmalar. Atatürk Üniversitesi Basimevi, 70 pp.

Pamilo, P., Varvio-Aho, S. & Pekkarinen, A. (1978) Low Enzyme Gene Variability in Hymenoptera as a Consequence of Haplodiploidy. Hereditas, 88, 93–99. http://dx.doi.org/10.1111/j.1601-5223.1978.tb01607.x

Pamilo, P., Pekkarinen, A. & Varvio, S. (1987) Clustering of Bumblebee Subgenera based on Interspecific Genetic Relationships (Hymenoptera, Apidae: Bombus and Psithyrus). Annales Zoologici Fennici, 24, 19–27.

Pamilo, P., Tengö, J., Rasmont, P., Pirhonen, K., Pekkarinen, A. & Kaarnama, E. (1997) Pheromonal and enzyme genetic characteristics of the Bombus lucorum species complex in Northern Europe. Entomologica Fennica, 7, 187–194.

Pawlikowski, T. (1996) Keys for the Identification of Polish Insects, Bees. Apidae. Subfamily: Apinae. Turpress. Polskie Towarzystwo Entomologiczne, 148 Serii, 1–56.

Pawlikowski, T. (1999) A Field Guide to Identification of Bumblebees (Hymenoptera: Apidae: Bombini) in Poland. Wydawnictwo Uniwersytetu Mikolaja Kopernika. Torun, 30 pp.

Pawlikowski, T. (2001) Computer Checklist of Apoidea (Hymenoptera) in Poland Part 1 (Excluding Bibliography to Distribution). Acta Universtatis Nicolai Copernici Biologia, 55 (107) 39-66.

Pekkarinen, A. & Teräs, I. (1977) Suomen Kimalaisista Ja Loiskimalaisista. Luonnon Tutkija, 81, 1–24.Pekkarinen, A. (1979) Morphometric, Colour and Enzyme Variation in Bumblebees (Hymenoptera, Apidae, Bombus) in

Fennoscandia and Denmark. Acta Zoologica Fennica, 158, 1–60.Pekkarinen, A., Varvio-Aho, S. & Pamilo, P. (1979) Evolutionary Relationships in Northern European Bombus and Psithyrus

species (Hymenoptera, Apidae) Studied on the Basis of Allozymes. Annales entomologici fennici, 45 (3), 77–80.Pekkarinen, A. & Teräs, I. (1993) Zoogeography of Bombus and Psithyrus in Northwestern Europe (Hymenoptera, Apidae).

Annales Zoologici Fennici, 30, 187–208.Pittioni, B. (1937) Die Hummelfauna des Kalsbachtales in Ost-Tirol. Ein Beitrag zur Ökologie und Systematik der Hummeln

Mitteleuropas. Festschrift zum 60. Geburtstag von Professor Dr. Embrik Strand, 3, 64–115. Pittioni, B. (1938) Die Hummeln und Schmarotzerhummeln der Balkan-Halbinsel. Die Hummeln und Schmarotzerhummeln

der Balkan-Halbinsel. II. Mitteilungen aus den Königlichen naturwissenschaftlichen Instituten in Sofia, 11, 12–69.Pittioni, B. (1939) Die Hummeln und Schmarotzerhummeln der Balkan-Halbinsel. II. Mitteilungen aus den Königlichen

naturwissenschaftlichen Instituten in Sofia, 12, 49–115.Radoszkowski, O. (1884) Révision des armures copulatrices des mâles du genre Bombus. Byulletin' Moskovskogo obshchestva

ispytatelei prirody, 59, 51–92.Rasmont, P. (1983) Catalogue Commenté des Bourdons de la région Ouest-Paléarctique (Hymenoptera, Apoidea, Apidae).

Notes Fauniques de Gembloux, 7, 71 pp.Rasmont, P., Terzo, M., Aytekin, M.A., Hines, H., Urbanova, K., Cahlikova, L. & Valterova, I. (2005) Pheromone analysis of

the bumble bee subgenus Sibiricobombus Vogt suggests Bombus niveatus Kriechbaumer and Bombus vorticosusGerstaecker are conspecific (Hymenoptera, Apidae, Bombus). Apidologie, 36, 571–584.

Zootaxa 3737 (2) © 2013 Magnolia Press · 181SUBGENUS THORACOBOMBUS (APIDAE, BOMBUS) IN TURKEY

TERMS OF USE This pdf is provided by Magnolia Press for private/research use. Commercial sale or deposition in a public library or website is prohibited.

Rasmont, P.,Aytekin, A.M., Kaftanoğlu, O. & Flagothier, D. (2009) Atlas Hymenoptera. Available from: http://zoologie.umh.ac.be/hymenoptera/page.asp?id=103 (Accessed 1 April 2013)

Reinig, W.F. (1967) Zur Kenntnis der Hummelfaunen einiger Gebirge West-Kleinasiens (Hymenoptera, Apidae). Nachrichtenblatt der Bayerischen Entomologen, 9 (10), 81–91.

Reinig, W.F. (1968) Über die Hummeln und Schmarotzerhummeln Nordwest-Anatoliens (Hymenoptera, Apidae). Nachrichtenblatt der Bayerischen Entomologen, 6, 101–112.

Reinig, W.F. (1971) Zur Faunistik und Zoogeographie des Vorderen Orients 3. Beitrag zur Kenntnis der Hummeln und Schmarotzerhummeln Anatoliens (Hymenoptera, Apidae). Veröffentlichungen der Zoologischen Staatssammlung München, 15, 139–165.

Reinig, W.F. (1973) Faunistische und Zoogeographische Studien in Kleinasien 4. Beitrag zur Kenntnis der Anatolischen Hummeln (Bombus Latr., 1802) und Schmarotzerhummeln (Psithyrus Lepeletier., 1832; Hymenoptera, Apidae). Mitteilungen der Münchner Entomologischen Gesellschaft, 69, 111–133.

Reinig, W.F. (1974) Faunistische und Zoogeographische Studien in Kleinasien 5. Auf Hummelfang im Taurus (Bombus Latr., 1802 et Psithyrus Lep., 1832; Hymenoptera, Apidae). Sonderabdruck aus dem Nachrichtenblatt der Bayerischen Entomologen, 23 (4), 67–80.

Richards, O.W. (1968) The Subgeneric Divisions of the Genus Bombus Latreille (Hymenoptera: Apidae). Bulletin of the British Museum (Natural History) (Entomology), 22, 210–276.

Robertson, C. (1903) Synopsis of Megachilidae and Bombinae. Transactions of the American Entomological Society, 29, 163–178.

Rohlf, F.J. & Slice, D.E. (1990) Extensions of the Procrustes method for the optimal superimposition of landmarks. Systematic Zoology, 39, 40–59. http://dx.doi.org/10.2307/2992207

Rohlf, F.J. (2000) Ntsys-Pc Version 2.1. Numerical taxonomy and multivariate analysis system. Exeter Software. Available from: http://life.bio.sunysb.edu/morph/ (Accessed 10 August 2013)

Rohlf, F.J. (2003) TpsSMALL1.20. Department of Ecology and Evolution, State University of New York at Stony Brook. Available from: http://life.bio.sunysb.edu/morph/ (Accessed 10 August 2013)

Rohlf, F.J. (2004) TpsSUPER1.14. Department of Ecology and Evolution, State University of New York at Stony Brook. Available from: http://life.bio.sunysb.edu/morph/ (Accessed 10 August 2013)

Rohlf, F.J. (2007) TpsRELW1.45. Department of Ecology and Evolution, State University of New York at Stony Brook. Available from: http://life.bio.sunysb.edu/morph/ (Accessed 10 August 2013)

Rohlf, F.J. (2008) TpsDIG2.12. Department of Ecology and Evolution, State University of New York at Stony Brook. Available from: http://life.bio.sunysb.edu/morph/ (Accessed 10 August 2013)

Rohlf, F.J. (2009) TpsUTIL1.44. Department of Ecology and Evolution, State University of New York at Stony Brook. Available from: http://life.bio.sunysb.edu/morph/ (Accessed 10 August 2013)

Skorikov, A.S. (1914) Les formes nouvelles des bourdons (Hymenoptera, Bombidae) VI. Russkoe éntomologicheskoe Obozrênie, 14, 119–129.

Skorikov, A.S. (1923) Palaearctic bumble bees. Part I. General biology (including zoogeography). Izvestiya Severnoi oblastnoi stantsii zashchity rastenii ot vreditelei, 4, 1–160.

Slice, D.E. (2007) Morpheus. Software for morphometric Research. Department of Ecology and Evolution, State University of New York, Stony Brook, New York. Available from: http://life.bio.sunysb.edu/morph/ (Accessed 10 August 2013)

Terzo, M., Valterova, I., Urbanova, K. & Rasmont, P. (2003) De la nécessité de redécrire les phéromones sexuelles des mâles de bourdons (Hymenoptera: Apidae, Bombini) publiées avant 1996 pour leur utilisation en analyse phylogénétique. Phytoprotection, 84, 39–49. http://dx.doi.org/10.7202/007806ar

Terzo, M., Valterova, I. & Rasmont, P. (2007) Atypical secretions of the male cephalic labial glands in Bumblebees: The case of Bombus (Rhodobombus) mesomelas GERSTAECKER (Hymenoptera, Apidae). Chemistry & Biodiversity, 4 (7), 1466–1471. http://dx.doi.org/10.1002/cbdv.200790124

Urbanová, K., Valterová, I., Rasmont, P. & Terzo, M. (2002) Znaèkovací feromony: slovení sekretu labiální vlázy samcù Bombus magnus (Marking pheromones of bumblebees: composition of the labial gland secretion of males of Bombus magnus). Chemické listy, 96, 918–919.

Vogt, O. (1911) Studien über das Artproblem. 2. Mitteilung. Über das Variieren der Hummeln. 2. Teil. (Schluss). Sitzungsberichte der Gesellschaft naturforschender Freunde zu Berlin, 31–74.

Wappler, T., De Meulemeester, T., Aytekin, A.M., Michez, D., Engel, M. (2012) Geometric morphometric analysis of a new Miocene bumble bee from the Randeck Maar of southwestern Germany. Systematic Entomology, 37, 784–792. http://dx.doi.org/10.1111/j.1365-3113.2012.00642.x

Williams, P.H. (1985) A preliminary cladistic investigation of relationships among the bumble bees (Hymenoptera, Apidae). Systematic Entomology, 10, 239–255. http://dx.doi.org/10.1111/j.1365-3113.1985.tb00529.x

Williams, P.H. (1994) Phylogenetic relationships among bumble bees (Bombus Latr.): a reappraisal of morphological evidence. Systematic Entomology, 19, 327–344. http://dx.doi.org/10.1111/j.1365-3113.1994.tb00594.x

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Williams, P.H. (2007) The distribution of bumblebee colour patterns world-wide: possible significance for thermoregulation, crypsis, and warning mimicry. Biological Journal of the Linnean Society, 92, 97–118. http://dx.doi.org/10.1111/j.1095-8312.2007.00878.x

Williams, P.H., Cameron, S.A., Hines, H.M., Cederberg, B. & Rasmont, P. (2008) A simplified subgeneric classification of the bumblebees (genus Bombus). Apidologie, 39, 46–74.

Williams, P.H., Tang, Y., Yao, J. & Cameron, S.A. (2009) The bumblebees of Sichuan (Hymenoptera: Apidae, Bombini). Systematics and Biodiversity, 7 (2), 101–190. http://dx.doi.org/10.1017/s1477200008002843

Zeldtich, M.L., Swiderski, H.D., Sheets, H.D. & Fink, W.L. (2007) IMP. Integrated Morphometrics Package. Available from: http://life.bio.sunysb.edu/morph/ (Accessed 10 August 2013)

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