A contribution to the vascular flora of Bulgaria: new species record and chorological update

: The present paper discusses new data on the distribution of 15 plant taxa, native or alien to the vascular flora of Bulgaria. Allium urusakiorum is reported to Bulgaria for the first time. Five of the listed taxa, Thelypteris palustris, Isolepis supina, Cladium mariscus, Carex punctata, and Avena eriantha are of conservation significance at regional scale. The current invasive status of the alien to the Bulgarian vascular flora Mollugo verticillata is discussed. Additional nomenclature notes and comments on some neglected herbarium records distributed in the indexed Bulgarian herbaria were included to the text.


Introduction
Floristic studies give an important information for plant biodiversity and are directly related to the protection of rare and threatened plant species, as well as their associated communities.In this regard, the Herbarium as an institution plays an important role for storing, managing, and distributing references for the chorology of plants of a certain geographic region.The Herbarium at the National Museum of Natural History at the Bulgarian Academy of Sciences, registered in the Index Herbariorum under the acronym BNHM is currently involved in the Distributed System of Scientific Collections -Bulgaria project (DiSSCo-BG; https://dissco-bg.eu/en�).The vascular plant collection of the herbarium BNHM is registered in the Global Registry of Scientific Collections with the code BG-NMNHS-BOT: https://scientificcollections.gbif.org/�.As part of the project up to now about 2000 herbarium exsiccates have been digitised through Specify 7 database management tool (https://www.specifysoftware.org/�).Among them, about 300 exsiccates concern recently collected specimens by the author from the territory of Bulgaria and Greece.In the present paper new regional chorologi-cal information, regarding 15 plant taxa is extracted from the corresponding collection.

Materials and methods
The new chorological information have been collected during botanical trips taken in the vegetation seasons of 2023.The identity of the plant species was keyed out through Stoyanov et al. (2021), apart from A. urusakiorum being identified via Koçyiğit at al. (2016).The nomenclature follows POWO (2023), except for Isolepis supina, presented as in Kuzmanov & Kozhuharov (1964).Herbarium sheets from all of the listed species had been deposited to the BNHM herbarium.Duplicates for some of the taxa had been deposited also in SO and SOM herbaria (herbarium acronyms follows Thiers, 2023).All of the cited specimens were digitised as part of DiSSCo-BG project.The plant names are ordered as in multivolume edition of Flora Europaea (Tutin et al., 1964(Tutin et al., -1980)).The species distribution is presented according accepted floristic regionalisation (Jordanov, 1966).The habitat notes were based on author's personal observations.The invasive status of the species is given according to Richardson & al. (2000).All the images (Figs.[1][2][3][4] presented in the text were taken by the author during his field trips.

List of plants
1. Thelypteris palustris Schott (Thelypteridaceae) (Fig. 1) Voucher specimens: 1) The Black Sea Coast (North): Novo Oryahovo Village, Dolni Chiflik Municipality, Varna Province, 0 m, 42.98189°N, 27.89025°E, coll. date: 8 Oct 2022;leg. & det. G. Kunev (BNHM-000000001178); 2) The Danubian Plain: Tarnene Village, Pleven Province, in the marshes near former Oil Refinery 'Plama',68 m,43.39187°N,24.49965°E,coll.The species is rare in Bulgaria and thus of conservation significance.It is listed as 'Vulnerable' in the national Red List (Ivanova, 2009), and included in Annex 3 of the Bulgarian Biodiversity Act.Its distribution in the country has been recently reevaluated (Tzonev et al., 2022;Natcheva et al., 2023).The localities 1 and 2 in the header data of the present report had been already mentioned by Tzonev et. al (2022) and here additional reference to the herbarium records are given.The locality 3 is new for the respective taxon and its first for the floristic region of Black Sea Coast (South).Three subpopulations were recognised from a distance, looking from the road at the East margin of the Alepu Marsh, as they appear as large rusty coloured spots densely colonised by the species.The microsite population area varies between 200-500 m 2 .The floristic composition includes Solanum dulcamara L., Cyperus odoratus L., Typha angustifo-  River, 80 m, 41.428250°N, 23.314180°E, coll. date: 30 Sep 2023, leg. & det. G. Kunev (BNHM-000000001885).Kunev (2019) reported this alien weed for the first time for Bulgarian flora following the find of only two specimens at the gravel bars of Struma River.The invasive status of the species in the country at that time was evaluated as 'casual alien' in the sense of Richardson et al. (2000).However, this particular section of the river was revisited each year since its report with attempts to collected additional data on population trends of the species.Furthermore, new localities were reported from Bulgaria and Greece (Kunev, 2020;Giannakis et al., 2022).The current report adds new localities at Struma River with numerous individuals unevenly spread over the gravel bars.This summarised information shows expansion of the species and increase in its population density in the region.Therefore, it could be concluded that there is persistent seed bank already established giving rise to many self-supported micro-populations, so the invasive status of the species should be reassess as 'naturalised alien'.
A new locality and first collection of the species from the Black Sea Coast (South) floristic region (Stoyanov et al., 2021) The taxon was reported to Bulgarian flora under the combination Stellaria media subsp.cupaniana (Jord.& Fourr.)Nyman (Cheschmedzhiev, 1988).It was first collected at Borovets, a mountain resort at Rila Mts, at approximately 1200-1300 m.However, the material at SOM-144911 (16 Sep 1984, leg. & det. I. Cheschmedzhiev) (incorrectly cited in Cheschmedzhiev, 1988 as 144901), is certainly referable to S. aquatica (L.) Scop.since its flowers bear five styles (vs.three in S. media subsp.cupaniana).
Under the combination Stellaria cupaniana (Jord.& Fourr.)Bég., it has been reported also for the territory of 'Rila Monastery' Nature Park and 'Rilomanastirska Gora' Reserve (Tsoneva & Peev, 2003;Tonkov et al., 2006), although not supported with herbarium collections in any of the national herbaria.In Bulgaria, up to now it has been known only from Rila Mts.The current report gives information for a new locality and a new floristic region for the species in the country.Two microsites composed of not more than 20 ind. each were observed at ruderal place along a dirt road.This is a hardly noticeable dwarf annual species, seldom collected in Bulgaria.It is characteristic species for Isoëto-Nanojuncetea communities, an ephemeral cyperaceous vegetation developed on silt depositions of temporary flooded stream sides and pond shores (Mucina et al., 2016) The species has been reported from Osogovo Mts (Ančev, 1982), thought without representative material deposited in any of the national herbaria.Probably, this was the reason for its omission in the latest floristic summaries from Bulgaria (Assyov & Petrova, 2012;Stoyanov et al., 2021).The abovecited material confirms the species for Osogovo Mts.The observed population consisted of about 30 individuals over 10 m 2 at the lowest point of a dry ravine, at its side slopes, under mixed beech-pine forest.
A. urusakiorum belongs to the sub-group of white-yellowish flowered members of the A. saxatile group.It was first reported from Strandzha Mts in European Turkey as A. saxatile M. Bieb.(Özhatay et al., 2012).However, further reevaluation proved the species as new to science (Koçyiğit at al., 2016).It is clearly separated from the above-mentioned taxa by its white-yellowish tepals, yellow anthers, which do not change colour over maturation, and longer filaments (Fig. 2a-b).
White-yellowish forms of A. saxatile were reported from Bulgarian parts of Strandzha Mts by Jordanoff (1934Jordanoff ( , 1935)).His materials were collected approximately 20 km from locus classicus of A. urusakiorum in Turkey and deposited at SO-11910 (15 Jul 1935, leg. & det. D. Jordanoff), -11911 (13 Aug 1933, leg. & det. D. Jordanoff).In August 2023, some of the localities reported by Jordanoff were revisited by the present author for observations and new samples to be made.The collected material were compared following the morphological description and the identification key in Koçyiğit at al. (2016), leading to the conclusion that the Bulgarian samples are undistinguishable from those described from Turkey, thus the name A. urusakiorum must be applied to the Bulgarian populations (sub A. saxatile) know from Strandzha Mts.
It must be mentioned that the tepals of A. urusakiorum collected in Bulgaria turned pinkish during the course of preservation, therefore their typical colour should be observed on fresh material.The species is seldom collected in the country, as it is confirmed by the low number of herbarium records kept at the SO, SOM and SOA herbaria.For this reason, it was included in the first edition of the Red Data Book of Bulgaria (Markova, 1984), as 'Endangered', and it is currently listed in the Annex 2a of the Biodiversity Act of Bulgaria.Most of the species records were documented before 1941 (Markova, 1984), with a single recent collection from the old riverbeds of Osam River at Balgarene Village, Pleven Province (Tzonev et al., 2010).Distribution notes on four new localities are here provided.The populations of I. supina at the reservoir at Staro Zhelezare Village, at the W shore of Brezevo Reservoir, and Maglene Reservoir consist of several tens to more than a hundred of individuals, while at the shores of Arkata Reservoir only one specimen has been noticed.Following the present and the historical records (Markova, 1984;Tzonev et al., 2010;Stoyanov et al., 2021) the species is now documented from seven floristic regions of Bulgaria: The Danubian Plain, North-Eastern Bulgaria, Sredna Gora (East), The Thracian Lowland, Tundzha Hilly Country, Vitosha Region, and The Rhodopes (East).The vertical limits of the species in the country is also significantly extended and exceeds 700 m at Arkata Reservoir.The species is widespread in the country, however no materials from Verila Mts has been collected up to now (Assyov & Petrova, 2012;Stoyanov et al., 2021) The species has been reported to Bulgaria for the first time from the region of Cape Kaliakra, on the Black Sea coast by Delipavlov (1999).Since it has been known from this single region, the species was evaluated as 'Criticaly Endangered' for the national Red Data Book (Tzonev, 2015a) and included in Annex 2a of the Bulgarian Biodiversity Act.
A single population of several hundreds of individuals were observed at Likana locality, over open, shallow, skeletal substrates, on slopes of low inclination and SE exposition.This represents the first observation of the species from the region of East Rhodope Mts (Stoyanov et al., 2021).The species is closely related with Avena clauda Durieu, the latter being reported from Belopolyane Village (Velchev & Petrova, 2011), very close to the here reported locality of A. eriantha.The latter can be distinguished by the spike with 2(-3) florets (vs.3(-5) in A. clauda) and the type of dispersal unit, being the spike (rachilla disarticulates at the base), but the floret in A. clauda (rachilla disarticulates between florets) (Delipavlov, 1999).
In the present report, the two taxa are considered at specific level.However, the taxonomic significance of the above-mentioned discrimination characters were not universally accepted.Some authors prefer merging A. eriantha and A. clauda, and treating them as different subunits of a single species, since both often co-occur across their range, interbreed, and produce viable offspring (Ladizinsky, 2012).The species is scattered and of conservation significance in Bulgaria being included in Annex 2a of the Bulgarian Biodiversity Act and in the Red Data Book of Bulgaria with the status of 'Endangered' (Stoeva, 2015).Additionally, the fens with Cladium mariscus are ascribed to the habitat 7210* of the Habitats Directive, and as an extremely rare communities in Bulgaria (Tzonev, 2015b) were included in Annex 1 of the Bulgarian Biodiversity Act.Therefore, this new locality at Marikostinovo is of great importance for the protection of the species in the country.
The present population has been observed in 2020 by Zhivko Barzov (an advanced nonprofessional naturalist), who shared his observation by a personal communication with the author.At the reported locality, the species forms dense patches at the source of the mineral spring, with monodominant stands covering at least 200 m 2 .The total area occupied by the species is most probably larger however, it was not fully assessed during current field study due to the restricted access to some of the zones of the balneological centre.The species is documented mostly from East Bulgaria (Valev, 1964).Here, a new locality and first  (Hájek et al., 2005;Hájek et al., 2007;Tzonev et al., 2013;Stoyanov et al., 2021).An additional collection of the species from Slavyanka Mts -SOM-151060 (9 Aug 1991, leg.I. Pashaliev, sub C. helodes Link., rev.: P. Jiménez-Mejias) was apparently neglected in the main floristic sources from Bulgaria, and as far as it is known was not reported elsewhere.
At the new locality at East Rhodopes Mts, the species occupies sides of temporary streamlet along with Anthoxanthum odoratum L., Ranunculus sardous, Trifolium nigrescens Viv., Juncus articulatus, Cynosurus cristatus L., Mentha aquatica, Persicaria hydropiper, Eleocharis uniglumis (Link) Schult., and Isolepis setacea (L.) R.Br.The population is particularly vulnerable since it is composed of less than hundred individuals and it is subject of grazing.This is rare sedge in Bulgaria, since it was reported from only few localities.It was included in the first edition of the Red Data Book of Bulgaria as 'Rare' (Meshinev, 1984), and currently it is listed in Annex 2a of the Bulgarian Biodiversity Act.Up to now, observations of the species has been made at the following regions of the country: Balkan Range (Central) -SOM-161680 (2 Jul 2004, leg. & det. M. Hajek et al.); Rhodopes Mts (Central) -SOM-79598

Introduction
Genus Theophilea (Cerambycidae) with type species T. cylindricollis Pic, 1895, was described by Pic in 1895 on the base of material from Armenia and subsequently it was reported from South European Russia, Ukraine and Hungary.After a comparative study of a large number of specimens Hladil (1988) considered that the specimens from Armenia are different from those collected in Hungary and described a new species T. subcylindricollis Hladil, 1988.Now the range of type species is limited to Armenia, Georgia, Turkey and Iran (Hoskovec et al., 2023).Currently, the genus Theophilea includes only these two species.
In 2023, a single specimen of T. subcylindricollis was observed and collected in a private yard in Vinarovo Village, Vidin Province.This note reports a second finding of the species in Bulgaria.

Methods
The specimen was photographed ex situ with an EOS 1200D (Canon) digital camera.The collected specimen was deposited in the collection of National Museum of Natural History in Sofia, Bulgaria.
Morphological notes: Theophilea subcylindricollis is apparently similar to Calamobius filum (Rossi, 1790), but it can be distinguished from the latter by the presence of long dark hairs on the ventral side of the antenna, the metallic luster of the elytra and the absence of a furrow on the median tibia (Bense, 1995;Dascǎlu, 2005).
Notes on biology: The larvae develop in the stems of grasses (Poaceae), e.g.Elymus repens, Poa angustifolia and Dactylis glomerata (Pil & Perić, 2012).The adults are active from April to July (Hoskovec et al., 2023) and are weak fliers (Pil & Perić, 2012).The habitats of the species are herbaceous communities in the flooded areas of the forest-steppe region, real steppes and loess steppes (Pil & Perić, 2012).
Notes on distribution: According to Zamoroka (2017) the original range of T. subcylindricollis was restricted to Pannonia and North Black Sea Region, but recently it is expanding westward, northward and eastward.According to Hoskovec et al. (2023) T. subcylindricollis is known from the Czech Republic, Hungary, North Macedonia, Moldova, Romania, Russia, Serbia, Slovakia, Ukraine and Kazakhstan.The species is also reported from Austria (Wiesbauer, New record of Theophilea subcylindricollis Hladil, 1988 in Bulgaria 2015), Bulgaria (Siering & Beier, 2019) and most recently from Albania (Kovács & Mesaroš, 2021).The species is rare and strictly protected in Serbia (Pil & Perić, 2012).
Both localities where T. subcylindricollis was recorded in Bulgaria are from the Danubian Plain, Northern Bulgaria, but are far away from each other.In Bulgaria the species was collected for the first time on 6 May 2017 around Ivanovski Manastir, Rusenski Lom River Valley, district of Ruse, Northeastern Bulgaria, and was subsequently reported by Siering & Beier (2019).The new locality in Vinarovo Village, Vidin Province is by far the northernmost in the country.Also there is a record from Gamzigradska Banja near Zaječar, Eastern Serbia (Popović et al., 2013), which is close to the border with Bulgaria and not far away from the new locality.
As the species was recently discovered in Bulgaria, further research is needed to establish its exact distribution in the country.It is possible that T. subcylindricollis can be found in other localities, most likely in Northern Bulgaria.

Introduction
According to the summarising paper of Kirçakci & Kabalak (2021) the total number of Turkish jewel beetle fauna is 421 species, which belong to 37 genera and 6 subfamilies.From a zoogeographic point of view, this fauna is most similar to the Greek and Bulgarian ones (Kirçakci & Kabalak, 2021).Later Volkovitsh et al. (2023) reported one species and one subspecies that are new for the Turkish fauna, and Tonğa & Sakalian (2023) added another new species for this country.
The aim of this study is to report new data on the distribution of 58 species and subspecies of jewel beetles in the European and Asiatic parts of Türkiye.A short key, which illustrates the main morphological characters of Agrilus (Quercuagrilus) graminis Kiesenwetter, 1857 and A. (Q.) hastulifer (Ratzeburg, 1837), which allows their better identification, is additionally provided.

Material and methods
The specimens were collected by traditional entomological methods such as: collecting the specimens from flowers and bushes by entomological net; shaking and beating of tree branches and crowns and collecting with Malaise traps during the period April-July 2007 and 2008.
The SEM photomicrographs were obtained using a JEOL GSM-5510, operating at 10 kV, the images were produced with a secondary electron detector at high vacuum.The magnification used to take each photomicrograph is shown in the corresponding photo.
The names of tribes, genera, subgenera, species and subspecies are listed in alphabetical order.The studied specimens are kept in the collection of the first author, Vladimir Sakalian (Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences), unless otherwise stated.
The Buprestidae fauna in Türkiye stands out as one of the richest and diverse within the Palearctic region.The unique zoogeographical structure of the country is shaped by a confluence of European, Mediterranean, and Middle Asiatic faunal elements.Although this fauna has been extensively studied in Türkiye, the continuous potential for the discovery and description of new taxa remains promising.
In the present work we report several new localities of the species in SE Bulgaria, as well as a new data on the host plants of the species.

Methods
The material for the present study was collected by the authors in the period 2018 -2023 from Eastern Rhodopes Mts, Maritsa River Valley and Strandzha Mts (Fig. 1).Adult beetles were hand collected from the host plants.The pictures were taken by using digital camera Olympus SP-820UZ (Fig. 1C), Canon PowerShot SX420 IS (Fig. 1A, B, D-F, Fig. 2) and a combination of Canon EOS 2000D digital camera, PRO-CA Camera Adapter, and a microscope Olympus SZ61 (Figs 3-6).The collected specimens are preserved in the Zoological Collection of Sofia University, Faculty of Biology (BFUS).riventris from Bistrets (Georgiev et al., 2005) was not found in the vicinity of this village during the present study, as well as nowhere else in the areas visited.
According Holzschuh (1984) and Bense (1995), the longitudinal rows of ocher (yellowish-brown) hairs on the elytra in both sexes, median tooth-like processes on the abdominal sternites 1 and 2 in males as well as the regularly sharpened apex of penis are among the main diagnostic features of P. serriventris (Figs 3, 4, 5A, 6A).In some examined specimens of P. serriventris, however, the longitudinal rows on the elytra are less pronounced to almost invisible.On the other hand, such rows are well developed in some of the P. tigrina specimens in the populations of the Western Stara Planina Mts (Gradinarov, 2016;Gradinarov & Petrova, 2019), as well as in populations of P. tigrina in Romania (Crișan et al., 2017).Among the males of P. tigrina from the locality of the species in the region of Beledie han (Gradinarov & Petrova, 2021) specimens with welldeveloped processes on the abdominal sternites 1 and 2 can also be found (Fig. 5B).Thus, the shape of the apex of penis is the most useful character for distinguishing P. serriventris (Fig. 6A, B) and P. tigrina (Fig. 6C, D).The pygidium in males of P. serriventris is significantly wider distally than in P. tigrina, which can also be used in distinguishing the two species (Holzschuh, 1984).
Roots of A. procera with live Cerambycidae larvae were collected on 9 September 2021, and April  16, 2023, from the locality near Lyubimets and that near Kotlari Vill., respectively.In both cases, single specimens of Phytoecia (Opsilia) coerulescens (Scopoli, 1763) (Cerambycidae: Lamiinae) have emerged from the roots after laboratory rearing.The last species is polyphagous mostly in Boraginaceae (Sama, 2002) and has been reported together with P. tigrina on A. barrelieri by earlier authors (Kovács, 2005;Crișan et al., 2017).
In our study, adults of P. serriventris were found on the host plants in the period of the second half of May to the beginning of June.Our data is consistent with the known period of activity of the adults of this species (Holzschuh, 1984;Georgiev et al.;2005, Fig. 5. Abdominal sternites in males of Pilemia spp.from Bulgaria.A -P. serriventris, Kotlari Vill., 27.v.2018;B -P. tigrina, Beledie Han Settlement, 20.v.2020.Scale bar: 1 mm.Hoskovec et al., 2023).Adults of P. serriventris appear to become active later in spring than those of P. tigrina, which in Bulgaria can be found on food plants already in April (Gradinarov 2016;Gradinarov & Petrova, 2019;2021).The late appearance of the adults of P. serriventris is probably related with the late flowering of the host plant A. procera, in comparison to A. barrelieri.
During the survey we found the species at an altitude of about 50 to 170 m. a.s.l., on alluvial banks of rivers from the Aegean drainage basin (Maritsa Riv., Arda Riv., Biserska Reka Riv.) and from the Black Sea drainage basin (Sredetska Reka Riv.).Near Lyubimets and near the Prohod Vill., it was also found among roadside vegetation at a greater distance (up to about 120 m) from the river.Both habitat types belong to the linear habitats (with linear strips of vegetation) sensu Bennett (2003).The second species of the subgenus Pilemia in Bulgaria -P.tigrina, has been recorded from linear habitats as well (Gradinarov & Petrova, 2021).In such habitats natural vegetation may have remained intact and they can serve as corridors for the dispersal of plant and animal species (Bennett, 2003).The presence of P. serriventris along the rivers Arda and Maritsa is also possible in the neighboring areas of Greece and the European part of Türkiye.

9.
Juncus capitatus Weigel (Juncaceae) Voucher specimen: The Mesta Valley: Ilinden Village, Hadzhidimovo Municipality, on the moist shores of a small reservoir, 630 m, 41.460340°N, 23.830110°E, coll.date: 24 Jul 2023, lLeg.& det.G. Kunev (BNHM-000000001880).New location and new floristic region for this dwarf annual species (Goranova et al., 2013), observed on sandy alluvium at the shores of a small pond.The communities in which the species participates could be referred as Isoëto-Nanojuncetea vegetation type.It was observed along the shores of the pond in four microsites with several tens of individuals.10.Avena eriantha Durieu (Poaceae) Voucher specimen: The Rhodopes (East): Likana Protected area, Ivaylovgrad Municipality, in forest