﻿Review of Orchidaceae of the northern part of Kazakhstan

﻿Abstract We present a review of Orchidaceae Juss. of the northern part of Kazakhstan, within the steppe, forest-steppe and semi-desert habitats of the country (Pavlodar, northern Kazakhstan, Kostanay, Akmola, Aktobe, West Kazakhstan, partially Karaganda and East Kazakhstan regions). The investigation is based on herbarium materials, literature data and field observations. We examined material from the following herbarium collections: LE, MW, TK, MHA, SVER, KUZ, ALTB, AA, NUR, KG, KSPI, NS, NSK, MOSP, ORIS, PPIU, totalling 288 herbarium specimens. The paper presents data in the form of revision, focusing on orchids of the northern part of Kazakhstan. It is accompanied by maps indicating localities, notes on habitat preferences, phenology and conservation status. A total of 25 species of 16 genera were recorded, of which eight are included in the Red Book of Kazakhstan (2014). According to our data, we propose to enlarge the number of protected orchids by adding the following nine species: Corallorhizatrifida, Epipactisatrorubens, Gymnadeniaconopsea, Hammarbyapaludosa, Herminiummonorchis, Liparisloeselii, Malaxismonophyllos, Neottiacamtschatea and Spiranthesaustralis. The most widespread species in the studied region are Dactylorhizaincarnata, D.umbrosa and Epipactispalustris. The rarest species (single locality only) are Epipactisatrorubens, E.helleborine, Epipogiumaphyllum, Hammarbyapaludosa and Herminiummonorchis.


Introduction
Orchids are one of the largest families in the world, numbering, according to various data, from 28,000 to 30,500 species (Chase 2005;Chase et al. 2015;Christenhusz and Byng 2016;Hassler 2023). Due to human encroachment and climate change, as well as other factors, many orchid species are at risk of extinction (Fay 2018;Zizka et al. 2021). Eight  The diversity of Orchidaceae Juss. in Kazakhstan is low due to the prevalence of an arid climate with a rather harsh temperature regime in the cold period. According to the last vascular plant list of Kazakhstan by Abdulina (1999), there are 31 species of orchids from 18 genera in Kazakhstan. However, several recent additions prove that the diversity of orchids in Kazakhstan is insufficiently studied. The following taxa were discovered in Kazakhstan for the first time since 1999: Cypripedium × ventricosum Sw. (Kotuhov et al. 2009(Kotuhov et al. , 2018, Epipactis atrorubens (Hoffm.) Besser (Perezhogin et al. 2015), Hammarbya paludosa (L.) Kuntze, Neottia cordata (L.) Rich. (Kubentayev et al. 2021).
In recent years, the study of orchids of Kazakh Altai, which accounts for 22 species, has received particular attention (Danilova et al. 2020;Sumbembayev et al. 2020aSumbembayev et al. , b, 2021Sumbembayev et al. , 2022Sumbembayev et al. , 2023, but orchids are still poorly studied in the northern part of Kazakhstan. Orchid diversity in the neighbouring countries of Kazakhstan is variable. According to the latest data, there are 1,449 species in China (Zhang et al. 2015), 135 species in Russia (Efimov 2020), 26 species in Mongolia (Baasanmunkh et al. 2021), 10 species in Kyrgyzstan (Lazkov and Sultanova 2014) and nine species in Uzbekistan (Schreder 1941). Khapugin (2020), based on the synthesis of published data on the global distribution of orchids within designated conservation areas, noted the insufficient study of orchids in central and northern Asia as a whole.
Taxonomical and geographical data about orchids presented by Abdulina (1999) and earlier sources are largely outdated. Therefore, we undertook the task of preparing a new, detailed revision of this family for the flora of Kazakhstan. Taking into account that orchid family is notable for numerous rare and protected species, we have provided a detailed revision that includes lists of localities. These lists can subsequently be directly used in documents aimed at establishing the protection of the Kazakhstan flora.
The purpose of this study was to clarify the species diversity and distribution of orchids in the vast territory of the northern part of Kazakhstan, based on the revision of herbarium materials, data from literature and field observations.

Material and method
Kazakhstan is located in the centre of Eurasia and the current ranking by area is ninth in the world with 2,724,900 km 2 . The territory of Kazakhstan is ecologically diverse, there are important zonal boundaries, including one separating the cold-temperate and temperate territories of northern Eurasia from the warm-temperate and hot-temperate territories of the Ancient Mediterranean (Abdulina 1999).
The presented contribution covers the major part of the country with the exception of the mountainous areas and desert areas, which are very different from the rest of the country and it is necessary to review them separately. In the article, the distribution of separate taxa is given according to both floristic and administrative principles. The studied area includes eight of 14 administrative regions (Fig. 1): Pavlodar, North Kazakhstan, Kostanay, Akmola, Aktobe, West Kazakhstan and partially Karaganda and East Kazakhstan regions). The administrative division of Kazakhstan that succeeded in 2021 is being pursued. Floristic subdivision of the territory follows Pavlov (1956). According to the latter classification, the studied area includes the following floristic regions (further abbreviated "FR"): Aktyubinsk, Bukeev, Emba, Eastern Upland, Irtysh, Karkaraly, Kokchetav, Mugodzhary, Priaral, Prikaspiy, Semipalatinsk Pinery, Syrt, Tobol-Ishim, Turgay, Ulutau and Western Upland (Fig. 1).
The nomenclature of each taxon mostly follows "Plants of the World Online" (POWO 2022).
The conservation status of each species follows the Red Book of Kazakhstan (2014), which assumes three categories of rarity: I -a very rare, critically endangered species; II -a very rare species; III -a rare species with a shrinking range.
Distribution maps of individual species were prepared using ArcMap. Dubious localities (with "question-mark" in the text) are included on the maps as well.

Results and discussion
According to our data, 25 species of orchids from 16 genera are recorded in the northern part of Kazakhstan. Eight species are listed in the Red Data Book of Kazakhstan, of which four species are classified under the II category and four species under the III category.
The larger number of species in Kokchetav, Tobol-Ishim and Irtysh floristic regions is due to the presence of the more variable spectrum of habitats for orchids, including pine, deciduous and mixed forests, river valleys, sphagnum swamps, flood meadows etc. The Mugodzhary FR, which is also relatively rich in orchids (8 species), is located in the semi-desert zone of Kazakhstan; however, the Urkach and Ber-Chugur places ("place" stands here for the Russian word "urochishche", which is used for various vernacular toponyms and also for the names of the former settlements) are located here, which include extensive lowlands with birch-aspen forests and sphagnum swamps, a very rare type of habitat in Kazakhstan. The Urkach place is considered to be a unique remnant of fragments of northern vegetation that retreated to the north during dry interglacial times and are evidence of the former vegetation of the Mugodzhar Mountains (Rusanov 1948).
Emba FR and Prikaspiy FR, where only one species of orchids (Orchis militaris L.) was found, as well as Turgay FR and Priaral FR, where orchids were not found at all, represent desert and semi-desert zones of Kazakhstan, with high soil salinity. The small number of orchids in Ulutau FR (also one species, Dactylorhiza incarnata (L.) Soó (Figs 3A, 5B)), in our opinion, is due to the poor knowledge of this region. The distribution of the studied species by administrative regions is the following: Akmola -12 species, Pavlodar -13 species, Karaganda -10 species, Kostanay -10 species, East Kazakhstan -8 species, North Kazakhstan -7 species, West Kazakhstan -3 species and Aktobe -8 species (Table 1).
Currently, eight species of orchids growing in the northern part of Kazakhstan are included in the Red Book of Kazakhstan (2014): Cypripedium calceolus L.    Based on our research, we discovered a single herbarium specimen from the Mugodzhary FR. In our assessment, it appears to be Dactylorhiza ochroleuca (Wüstnei ex Boll) Holub. However, this finding requires confirmation, as there is a possibility of confusion with hypochromic variants of Dactylorhiza incarnata. Dactylorhiza sibirica Efimov (Figs 3C, 5D) is reported for the first time for the northern part of Kazakhstan. Many taxa are reported for the first time for particular floristic and administrative regions of the country.
Dactylorhiza majalis (Rchb.) P.F. Hunt & Summerh and D. russowii (Klinge) Holub, reported earlier for the studied region (Pavlov 1956;Aipeisova 2012Aipeisova , 2013, are excluded from the flora of Kazakhstan as erroneous determinations. More recently, Sumbembayev et al. (2023) reported Dactylorhiza × kerneri (Soó) Soó (= D. fuchsii × D. incarnata) for the flora of Kazakhstan, based on herbarium collections stored in LE. We believe that those specimens can be rather safely determined as Dactylorhiza sibirica, a hybridogenous species originating according to the same hybrid formula. Habitat and ecology. Forest swamps, stream valleys, lakesides, sphagnum swamps and wet birch forests.

Corallorhiza trifida
Phenology. Flowering in May-Jun; fruiting in Jul-Aug. Conservation status. Not protected. We recommend to include this species in the next edition of the Red Book of Kazakhstan.
Notes. Corallorhiza trifida is reported for the Kokshetau State National Nature Park for the first time. Habitat and ecology. Birch and birch-pine forests, forest stream valleys, forest swamps and forest lake shores.
Phenology. Flowering in Jun; fruiting in Jul-Aug. Conservation status. It is included in the Red Book of Kazakhstan (category III). It is a rare and endangered species protected in the "Burabay" State National Nature Park, "Sogrov" State Nature Reserve and "Floodplain of the Irtysh River" State Nature Reserve.
Notes. Some populations of Cypripedium calceolus are located near Shchuchinsk and the village of Burabay, in areas with high recreational activity. These populations require special attention and protection due to the low number of plants in the populations, which can be attributed to the significant anthropogenic impact in these areas (Sultangazina et al. 2014;.  (Pugachev 1994), ?Mendykara District (Pugachev 1994). Irtysh: Pavlodar Region [without detailed locality] (Kusnetsov and Pavlov 1958).

Cypripedium guttatum
Habitat and ecology. Wet birch forests. Phenology. Flowering in Jun; fruiting in Jul-Aug. Conservation status. This rare species is included in the Red Book of Kazakhstan (category II) and is protected within the territory of two State Nature Reserves: "Sogrov" and "Floodplain of the Irtysh River".
Notes. The report of Cypripedium guttatum for the Kostanay Region is doubtful since we have not found herbarium collections from these areas, including the herbarium of Kostanay Pedagogical University (KSPI), where the Pugachev collections are stored.  (Semenov 1928;Kusnetsov and Pavlov 1958;Gorchakovskiy 1987 Habitat and ecology. Birch forests and valleys of forest streams. Phenology. Flowering in Jun; fruiting in Jul-Aug. Conservation status. This very rare species is included in the Red Book of Kazakhstan (category II). It is protected within the "Sogrov" and "Floodplain of the Irtysh River" State Nature Reserves.

Cypripedium macranthos
Notes. According to recent reports (Sultangazina et al. 2014;) and our field studies, there is currently no confirmation of the presence of Cypripedium macranthos within the territory of Kokchetav FR.
One specimen of C. ×ventricosum (C. calceolus × C. macranthos) hybrid was found: "Tobol-Ishim: North Kazakhstan Region: Kyzylzhar District: on the right shore of the Ishim River, near Krasnoyarka, 17 Jun 1968, Tsykareva s.n. (AA!)". This species occurs in areas where parent species co-occur, forming transitional populations with intermediate morphology (Averyanov 1999;Knyazev et al. 2000;Filippov and Andronova 2011;Andronova et al. 2017). This hybrid is reported for the first time in the studied region; Previously it was only reported in the Katon-Karagai District of the East Kazakhstan region in Kazakhstan (Kotuhov et al. 2009(Kotuhov et al. , 2018. Habitat and ecology. Moist pine and birch forests, along the shores of forest streams and lakes, forest swamps.

Dactylorhiza fuchsii (Druce) Soó
Phenology. Flowering in Jun-Jul; fruiting in Aug-Sep. Conservation status. It is included in the Red Book of Kazakhstan (category II). It is protected on the territories of the "Naurzum" State Nature Reserve; "Kokshetau" State National Nature Park, "Burabay" State National Nature Park, Karkaraly State National Nature Park; Botanical nature monument "Planting of birch and pine forests near Borovskoye Lake".
Notes. During the revision of KUZ herbarium materials, we identified two specimens of D. fuchsii, that were previously incorrectly determined as Dactylorhiza russowii. Based on these incorrectly misidentified herbarium specimens, D. russowii was previously reported for the Kokchetav FR (near Burabay, Habitat and ecology. Wet meadows, flooded saline meadows, valleys of rivers, streams, lakeshores. Phenology. Flowering in Jun-Jul; fruiting in Aug-Sep. Conservation status. Not protected. Notes. Dactylorhiza incarnata is the most common orchid species in the studied region, found in almost all areas. However, for Turgay FR, it was recently reported by mistake. The corresponding localities (Sumbembayev et al. 2023) actually refer to the Tobol-Ishim FR. There are currently no known orchid species that occur in Turgay FR. Existing reports of D. traunsteineri (Saut. ex Rchb.) Soó for Kazakhstan are most likely based on misidentified D. incarnata. Therefore, the report of D. traunsteineri for Urkach Plateaeu in Mugodzhary (Kusnetsov and Pavlov 1958) is possibly based on a herbarium specimen in LE with incomplete label (Alexandri Lehmann Reliquiae botanicae, Orchis angustifolia, [det.] Al. Bunge), which was probably collected near the end of May 1842 (Bunge 1847).
During the revision of MHA herbarium materials, we found a herbarium specimen from the Mugodzhary FR (Aktobe Region: Mugalzhar District: the southern tip of the Mugodzhar Mountains, west of Algabas railway station, 21-22 Jun 1987, Skvortsov s.n. (MHA!)). In our opinion, this specimen corresponds to Dactylorhiza ochroleuca. The identification is based on information provided on the herbarium label ("pale, pink-fawn flowers"). However, it is possible that these plants belong to hypochromic variants of D. incarnata. Exact determination is possible only through allozyme analysis   Notes. Dactylorhiza maculata is often hardly distinguishable from D. fuchsii. When they co-occur, they form populations that include plants with intermediate morphology, indicating possible hybridisation. We consider that D. maculata is generally a European species, with only isolated occurrences in Asia, particularly in the western part of Siberia and in Kazakhstan. Determining the exact eastern distribution limit of this species is challenging due to its similarity with D. fuchsii in this region, where their ranges overlap.

Habitat and ecology. Lowlands amongst birch trees, damp saline meadows and floodplains of rivers.
Phenology. Flowering in Jun-Jul; fruiting in Aug-Sep. Conservation status. Not protected. Notes. Dactylorhiza salina is reported here for the first time for the Tobol-Ishim FR and Kostanay Region. D. salina was recently erroneously reported for the Turgay FR (Sumbembayev et al. 2023); in fact, the corresponding localities refer to the Tobol-Ishim FR. There are currently no orchid species known to occur in the Turgay FR. D. salina is hardly distinguishable from D. umbrosa. Notes. Dactylorhiza sibirica is reported for the studied region for the first time. This allopolyploid species was described relatively recently, in 2016, with diploid Siberian D. fuchsii and D. incarnata as its presumable parental taxa (Efimov et al. 2016). In eastern Kazakhstan, the species was earlier incorrectly determined as Dactylorhiza baltica (Klinge) Nevski or Dactylorhiza × kerneri (Danilova et al. 2020;Sumbembayev et al. 2023). Habitat and ecology. Valleys of rivers and streams, along the damp edges of birch and aspen forests, through swamps, salt marshes, wet meadows, in the lowlands amongst birch thickets.

Dactylorhiza sibirica Efimov
Phenology. Flowering in Jun-Jul; fruiting in Aug-Sep. Conservation status. Not protected. Notes. Aipeisova (2012Aipeisova ( , 2013 reported D. majalis. for Mugodzhary (near Ber-Chogur and Mount Boktybai). Although corresponding herbarium specimens were not located, we believe that this report is an obvious mistake. It is more likely that the plants were D. umbrosa, as we found herbarium materials collected from the same place (near Ber-Chogur, 10 Jun 1927, Rusanov s.n (AA!)). We consider D. umbrosa and D. salina to be closely-related species and determining plants with certainty can sometimes be challenging. Habitat and ecology. In mixed and deciduous shady forests, at the forest edges. Phenology. Flowering in Jun-Jul; fruiting in Aug-Sep. Conservation status. Not protected. It requires protection at the regional level. Notes. We did not find any herbarium collections of E. helleborine from the studied region and the existing literature reports require confirmation. However, this species is known to be common in mountainous areas of southern and eastern Kazakhstan (Kuznetsov and Pavlov 1958), which are not included in the current revision. Habitat and ecology. Marshy meadows, river valleys, in wet forests. Phenology. Flowering in Jun-Jul; fruiting in Aug-Sep. Conservation status. This species is included in the Red Book of Kazakhstan (category III). It is protected in the following territories: "Naurzum" State Nature Reserve, "Semey Ormany" State Nature Reserve, "Burabay" State National Nature Park, "Karkaraly" State National Nature Park, as well as the State Nature Reserves of "Floodplain of the Irtysh River", "Orkash", "Kokzhide-Kumzhargan" and "Budarinskiy". It is also protected in the natural monument "Birch and pine plantations forests near Borovskoye Lake". Epipactis palustris is one of the most widely distributed orchids in Kazakhstan. Currently, there is a need to reconsider the necessity of state protection for this species. Habitat and ecology. Swampy pine forests, swamps. Phenology. Flowering in Jul-Aug; fruiting in Sep-Oct. Conservation status. This species is included in the Red Book of Kazakhstan (category II) as a rare species found in small numbers within a limited area. In the studied region, it is preserved in the "Karkaraly Biological Reserve". Habitat and ecology. Sedge and sphagnum swamps. Phenology. Flowering in Jun-Jul; fruiting in Aug-Sept. Conservation status. Not protected. The species is extremely rare and we consider it mandatory to include it in the next edition of the Red Book of Kazakhstan.

Notes. The species in the studied region is known from a single locality in
Notes. Liparis loeselii is reported here for the first time for Semipalatinsk Pinery and Mugodzhary FRs. Previously, in the studied region, the species was observed only in Kokchetav FR (Karamysheva and Rachkovskaya 1973;Sultangazina et al. 2014;. The reported localities of L. loeselii belong to the type subspecies, whereas the locality in East Kazakhstan represents the newly-described L. loeselii subsp. Orientalis, which differs from the typical subspecies by having broader leaf blade, more visible petioles and broader fruits (Efimov 2010).
Specimens examined and literature records. Tobol-Ishim: Kostanay Region: Mendykara District: Borovskaya water protection forest dacha, 17 Jul 1925, Rusanov 1553 (LE!); near Borovskoye (Pugachev 1994); near Kamenskural'skoe (Pugachev and Masyukova 1969); Auliekol District: near Kalininskoye (Pugachev 1994 Habitat and ecology. Grassy birch forests, near lakes, along forest streams. Phenology. Flowering in Jun-Jul; fruiting in Aug-Sept. Conservation status. Not protected. We recommend to include this species in the next edition of the Red Book of Kazakhstan. Notes. Forms both with one and with two well-developed leaves occur in Kazakhstan, the latter being recognised as a variety of M. monophyllos var. diphyllos (Cham.) Luer (e.g. by Pavlov 1928). Bayan-Aul and Karkaraly Mts represent the southernmost locality of the species in Central Asia. Currently, field studies are needed to verify the presence of Spiranthes australis in the study region, since the species was not observed here for almost 30 years.