ACTA BOT. CROAT. 76 (2), 2017 191 Acta Bot. Croat. 76 (2), 191–195, 2017 CODEN: ABCRA 25 DOI: 10.1515/botcro-2017-0003 ISSN 0365-0588 eISSN 1847-8476 Short communication Bouché’s star of Bethlehem, Ornithogalum boucheanum (Kunth) Asch. (Hyacinthaceae), a new species in flora of Croatia Dragica Purger1*, Sanja Kovačić2, János Csiky1 1 Department of Ecology, Institute of Biology, Faculty of Sciences, University of Pécs, Ifjúság útja 6, H-7624 Pécs, Hungary. 2 Botanical Garden, Department of Biology, Faculty of Science, University of Zagreb, Trg Marka Marulića 9A, 10000, Zagreb, Croatia. Abstract – Populations of Bouché’s star of Bethlehem (Ornithogalum boucheanum (Kunth) Asch., fam. Hy- acinthaceae) were recorded on Bansko Hill (Baranja, Croatia) in 2007. Since this species has not been previ- ously confirmed in Croatia, it should be treated as a new taxon in the country and included in the Flora Cro- atica Database. In this paper we present a short morphological description of O. boucheanum and diagnostic morphological characters for differentiation from the related O. nutans L. We suggested O. boucheanum be evaluated as a critically endangered (CR) species of the Croatian flora, considering the small number of indi- viduals and the small extension of its population. The recording of its populations on the edge of the loess cliff in Bansko Hill a part of which belongs to the Important Plant Area and Natura 2000, confirms the sig- nificance of this unique habitat in preserving rare and endangered plants. Key words: Bansko Hill, Baranja, loess, Myogalum boucheanum, Ornithogalum nutans, threatened plants * Corresponding author, e-mail: dragica@gamma.ttk.pte.hu Introduction Bouché’s star of Bethlehem (Ornithogalum boucheanum (Kunth) Asch.; Österr. Bot. Zeitschr. (1866) 16: 192), (Syn.: Myogalum boucheanum Kunth, Honorius boucheanus (Kunth) Holub, Ornithogalum nutans L. subsp. boucheanum (Asch.) Hayek; Liliales, Hyacinthaceae), is a lowland-colline plant, an Eastern-sub-Mediterranean-Pannonic (Pontic) floristic element (Soó 1973). The species is distributed in Central and Southeastern Europe, where it is considered native, while in North and Western Europe it is treated as adventive (Zahariadi 1980). The species is distributed in the Cauca- sus, around the Black Sea, through Anatolia and the Bal- kans to the eastern part of the Danube valley and the Car- pathian Basin (Soó 1973). According to Jávorka (1925) it does not occur in Croatia. During a survey of the loess flora and vegetation on Bansko Hill in Baranja (NE Croatia) we found two small populations of Ornithogalum bouchea- num. This species is not included in the recent Croatian bo- tanical literature (cf. Domac 2002; Nikolić 2000; Nikolić and Topić 2005) or in the national database of the Croatian flora (Nikolić 2016a,b). There are no data about this species in the papers dealing with the flora of Bansko Hill (Sturc 1988) and Baranja (Panjković 1990, Zahirović 2000). The occurrence of a related species, the Nodding star of Bethle- hem (Ornithogalum nutans L.) is reported from the north- ern part of Croatia (Rauš and Šegulja 1983) and several Adriatic islands (Nikolić 2016a). The native range of this species is smaller than that of the related O. boucheanum and it comprises Anatolian and Balkan peninsulas. In most of Europe it is considered to be an alien species, or else it is of unknown status (Euro+Med 2006–). Ornithogalum boucheanum and O. nutans are two spe- cies of the subgenus Myogalum (Link) Baker in Europe spe cimens of which are difficult to separate on the basis of mor phological characters, especially after they have bloomed (Zahariadi 1980). Since some authors of the last century (e.g. Hayek 1933) considered O. boucheanum a subspecies of O. nutans (O. nutans L. subsp. boucheanum (Asch.) Ha- yek), there is a possibility, that these two taxa were not properly distinguished when collected in the last century. Therefore our aim was to check the specimens in Herbari- um Croaticum (ZA) and Herbarium in Budapest (BP) and to clarify the status of these two related taxa in Croatia. In this paper we present a short morphological descrip- tion of O. boucheanum and diagnostic morphological char- acters for differentiation from the related O. nutans. PURGER D., KOVAČIĆ S., CSIKY J. 192 ACTA BOT. CROAT. 76 (2), 2017 Material and methods For determination of plant species we used the recent Croatian (Domac 2002) and Hungarian (Király 2009) hand- books. The description of the species is given based on characteristics of the collected specimens and according to related literature (e.g. Speta 2008, Meriç et al. 2011). The distribution map of O. boucheanum in Croatia (Fig. 1) was compiled according to the Central European Flora Mapping System (Nikolić et al. 1998). Geocoding of site was made by a GPS device. For classification of O. boucheanum into IUCN red list categories we used criteria after Nikolić (2005) and IUCN (2012) amended for Croatia in 2014. Ex- amples of plants have been collected (on April 16, 2008) and a voucher specimen was deposited in Herbarium Cro- aticum (ZA). Study area Bansko Hill (Fig. 1.) is a 21 km long elevation (243 m a.s.l.) extending in a NE–SW direction, and consists of loess sediments. Its southeastern part is a sheer, 25–58 m high loess cliff, facing the floodplain of the River Danube (Purger and Csiky 2008). Results and discussion During a survey of loess flora and vegetation on Bansko Hill (NE Croatia) from Zmajevac to Batina (0178/4 Central European Flora Mapping quadrate) two small populations of O. boucheanum were recorded (Fig. 1.), at the very bor- der of the Natura 2000 site HR2001309 “Dunav North of Kopački rit” (Anonymous 2013a). WGS 84 coordinates, altitude, population size and brief habi- tat descriptions: 1. N 45°49’11.10”, E 18°49’40.14”, 100 m a.s.l., small pop- ulation of fewer than 10 individuals, on the edge of the pla- teau, in a secondary xerophilous forest with Fraxinus or- nus, Acer campestre, Robinia pseudacacia in the tree layer, Crataegus monogyna, Prunus spinosa, Viburnum lantana in the shrub layer and Anthriscus cerefolium, Ballota nigra, Bromus sterilis, Chelidonium majus, Erodium ciconium, Ga lium aparine, Geranium rotundifolium, Stellaria media and Veronica hederifolia in the herb layer (leg./det.: J. Csiky, D. Purger, 26.04.2007, Zmajevac); 2. N 45°49’13.99”, E 18°49’42.39”, 100 m a.s.l., small po- pu lation of fewer than 100 individuals, in a secondary xe- rophilous forest with Robinia pseudacacia, Fraxinus ornus in the tree layer, Crataegus monogyna, Prunus spinosa, Eu- onymus europaeus, Ulmus minor in the shrub layer, An- thriscus cerefolium, Artemisia campestris, Arum orientale, Ballota nigra, Bromus sterilis, Carduus acanthoides, Cheli- donium majus, Securigera varia, Erodium ciconium, Eu- phorbia cyparissias, E. helioscopia, Fumaria schleicheri, Galium aparine, G. mollugo, Geranium pusillum, Geum urbanum, Lamium purpureum, Melandrium album, Musca- ri neglectum, Onopordon acanthium, Sisymbrium orientale, Stellaria media, Veronica hederifolia, Vicia narbonensis subsp. serratifolia, V. pannonica, Viola cf. odorata in the herb layer (leg./det.: J. Csiky, D. Purger, 16.04.2008., Zma- jevac [ZA]). The populations on Bansko Hill occurred in the shrubs and transitional zone of grassland and forest in the very border of the loess plateau. These are the only known and proved localities of O. boucheanum in Croatia. Since this species has not been previously identified in the country, it should be treated as a new taxon of the vascular flora of Croatia and included in the Flora Croatica Database (Niko- lić 2016a). We also checked data of similar, related species O. nu- tans to clarify its status in the country. In the old botanical literature occurrence of this species (“between Vukovar and Sotin”) is mentioned (Strein ap. Schlosser and Vukotinović Fig. 1. Distribution of Ornithogalum boucheanum (*) on Bansko Hill (Croatia). A grid square measures approximately 6500 (latitudinal) ×5500 (longitudinal) metres. Abbreviations: Ba – Batina; BM – Beli Manastir; Br – Branjina; BV – Branjin Vrh; Dr – Draž; Ga – Gajić; Ka – Kamenac; Kr – Karanac; Ko – Kotlina; KV – Kneževi Vinogradi; Pd – Po- dolje; Pp – Popovac; Su – Suza; Zm – Zmajevac (drawn by J. Csiky and T. Nikolić). ORNITHOGALUM BOUCHEANUM THE NEW SPECIES IN FLORA OF CROATIA ACTA BOT. CROAT. 76 (2), 2017 193 Fig. 2. Ornithogalum boucheanum near Zmajevac on Bansko Hill: a) flowers and b) fruits (Photo: J. Csiky and D. Purger). 1869). The specimen of Star of Bethlehem collected by Pavich in the 19th century near Sava River (Slavonia), iden- tified as O. nutans is deposited in the Herbarium Croaticum (ZA). It was the only record of this species reported from the continental part of Croatia (Nikolić 2016a). After check- ing this specimen in ZA, we found that it was Ornithoga- lum umbellatum s.l. misidentified as O nutans. Probably upon these old data the Nodding star of Bethlehem (O. nu- tans) is listed in the Flora of Slavonia and Baranja (Rauš and Šegulja 1983), though without any localities or recent findings. Accordingly, we can conclude, that the distribu- tion of O. nutans in Croatia is restricted to the Mediterra- nean part of the country (Nikolić 2016a). There are three specimens in the Herbarium in Budapest which are sup- posed to be O. boucheanum and are said to originate from Croatia. On these sheets data about locality, habitat and date of collecting are missing or ambiguous, therefore upon that material we could not confirm the occurrence of O. boucheanum in the Mediterranean part of Croatia. Accord- ing to the data of its distribution in Hungary, it seems that this species becomes rare towards the south-west (towards Croatia) and it is already rare in Baranya County (Bartha and Király 2015). As the handbook for the identification of plants in Croa- tia (Domac 2002) does not contain O. boucheanum, here we provide a description of this species and additional in- formation for its determination. According to our own knowledge and literature data (Zahariadi 1980, Hrouda 2002, Mesterházy and Király 2009, Meriç et al. 2011), we summarised the morphological and anatomical features of O. boucheanum and the closely related O. nutans, which are very similar, but differ in several characters (Tab. 1). Ornithogalum boucheanum is a 20–60 cm high plant. Bulb ovoid-globose, 30×40 mm, tunica light brown. Scape erect, cylindrical, light green. Leaves (3–6) bright green, as long as the scape or shorter, 1.0–1.5 cm wide, 20–40 cm long, margin entire, edges are parallel and gradually taper- ing to acute apex, at flowering time withered. The adaxial face of the leaf has white median line. Raceme corymbose, dense, 10–20 flowered. Flowers scentless. Bract 22–25 mm long, lanceolate-acuminate, light brown and membrane-like structured. Pedicel 5–10 mm, 10–13 mm in fruit. Perianth segments 18–25 (–30) mm long, 2–3.5 mm wide, lanceo- late, milky-white with green fascia inside and outside (Fig. 2a). Anthers 2–3 (–3.5) mm, white, light yellow, filament 4–5 (–6) mm, lanceolate, wide and winged, ending with two lateral teeth. Stigma spatulate. Ovary ovoid-elliptic (green), as long as style (white). Capsule ovoid (Fig. 2b) 18×12 mm, erect. Seeds numerous, black, subglobose to globose, black and reticulate with wrinkled, undulate walls. According to the results of Bednorz and Czarna (2008) seeds of O. boucheanum and O. nutans, two closely related and morphologically very similar species, are practically undistinguishable. Although the species are similar in re- gard to the scape anatomy, their leaf anatomy is different: mesophyll in O. boucheanum is thicker, differentiates as palisade parenchyma which is present on both the adaxial and abaxial side and spongy parenchyma, which contains large lacuna (Meriç et al. 2011). Ornithogalum is a taxo- nomically difficult genus; its morphology is poorly corre- lated with the variation in chromosome number and karyo- type. The chromosome numbers of O. boucheanum have been determined as 2n=56 (basic chromosome number x= 8), while the chromosome numbers of O. nutans are deter- mined as 2n=14 and 2n=35 (basic chromosome number x= 7) (Dalgıc and Ozhatay 1997, Dalgıc et al. 2009). O. boucheanum rarely occurs in Southern Transdanubia (Hungary), in grasslands (Purger 2008) and spring weed vegetation in vineyards (Pál 2007); in this part of the coun- Tab. 1. Diagnostic characters of Ornithogalum boucheanum and O. nutans. Characters O. boucheanum O. nutans Raceme (5)10–15(20) flowered (3)6–10(12) flowered Perianth segments (length) 20–35 mm 15–20 mm Perianth segments (colour) green and white inner side, dark green stripe on outer side white inner side, greenish stripe on outer side Ovary (length) 4–7 mm 3–5 mm Style (length) 5–7 mm <5 mm Toothed filaments 6 3 Seed 2–2.5 mm, broadly ovoid-orbicular 1.5–2 mm, ovoid-orbicular Leaf anatomy mesophyll equifacial (1000 μm) mesophyll unifacial (700 μm) Pollen grains (P – polar axis, E – equatorial axis) P = 70.84±0.35 μm E = 50.27±1.22 μm P = 64.02±0.15 μm E = 42.01±0.20 μm PURGER D., KOVAČIĆ S., CSIKY J. 194 ACTA BOT. CROAT. 76 (2), 2017 try it is considered to be a strongly endangered plant (Pál 2005). In Hungary it grows in clay (loamy) soils on loess and in humic sandy soils and can be found in different habi- tats, mostly in forest clearings, sparse oak forests, on open slopes, in valleys along streams, in lowland-colline areas with altitudinal range up to 500 m (Soó 1973). According to Borhidi (1995) it is a natural weed, a plant of disturbed hab- itats which belongs to the vegetation of class Artemisietea. In Serbia it occurs only in the Pannonian part (Vojvodina province): the Deliblato sandy area, the vicinity of Kikinda (Diklić 1975), Riđica, Sombor, Bački Monoštor (Obradović et al. 1981); Fruška gora mountains: Irig (Butorac 1992). Focus on the Carpathian Basin: O. boucheanum is a sparse but not endangered species in Hungary (Király 2007), rare and critically endangered in Austria (Speta 2008). In Slovakia this species is endangered (EN) accord- ing to Feráková et al. (2001), however recently Eliáš et al. (2015) considered it as least concern (LC). In Serbia it is a statutorily protected but not threatened species (Stevanović 1999). So far, Ornithogalum boucheanum is known in Croatia from only the two (very close) localities presented here, its estimated ‘area of occupancy’ being less than 10 km2 (crite- rion B1a and B2a) and population number of the species is estimated to fewer than 1000 mature individuals (criterion C), therefore it could be evaluated as a critically endan- gered (CR) species of the Croatian flora (Nikolić 2005; IUCN 2012). Accordingly, we also propose that this species should be included among the statutorily strictly protected species of Croatian flora (acc. to Anonymous 2013b). Since loess cliff habitats are very rare and sporadically distributed in NE Croatia, a part of this area is considered an IPA (Important Plant Area) site and belongs to the Natu- ra 2000 network. 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