Elena I. Terentieva, Senior Researcher, Botanical Garden of the Lomonosov Moscow State University (Russia, 119991, Moscow, Leninskie gory 1, 12); e-mail:
Tatyana I. Varlygina, Senior Researcher, Botanical Garden of the Lomonosov Moscow State University (Russia, 119991, Moscow, Leninskie gory 1, 12); e-mail:
Galina F. Darman, Senior Researcher, Amur Branch of the Botanical Garden of the Institute, FEB RAS, (Russia, Amur Region, 675004, Blagoveshchensk, 2 km, Ignatievskoe Shosse); e-mail:
Galina V. Degtjareva, Leading Researcher, Botanical Garden of the Lomonosov Moscow State University (Russia, 119991, Moscow, Leninskie gory 1, 12); e-mail:
Sergey V. Efimov, Senior Researcher, Botanical Garden of the Lomonosov Moscow State University (Russia, 119991, Moscow, Leninskie gory 1, 12); e-mail:
Tagir H. Samigullin, Belozersky Institute of Physico-Chemical Biology of the Lomonosov Moscow State University (Russia, Moscow 119992, Leninskie gory 1, 40); e-mail:

Reference to article

Terentieva E.I., Varlygina T.I., Darman G.F., Degtjareva G.V., Efimov S.V., Samigullin T.H. 2020. Revision and distribution of Liparis species (Orchidaceae) in Amur region (Russia). Nature Conservation Research 5(Suppl.1): 102–113.

Electronic Supplement 1. Voucher information and GenBank accession numbers for taxa used in this study (Link)
Electronic Supplement 2. Phylogenetic relationship in Liparis as revealed by maximum parsimony analysis (Link)

Section Research articles

In July 2018, in the Khingan State Nature Reserve and Muravyevskiy Park of Sustainable Development, we conducted the study of Liparis populations and preliminary determining the orchid individuals collected. In the family Orchidaceae, Liparis is one of the most difficult genera in terms of systematics. Observations in nature, research of herbarium specimens and literature demonstrate the considerable intraspecies variability in morphological traits. Therefore, to assess the taxonomic position of orchids from natural populations, we collected parts of the plants for further molecular-phylogenetic research. In total, we included 110 samples in the analysis. As a molecular marker, we used the internal transcribed spacers (ITS1–5.8S–ITS2) of 18S–26S nuclear ribosomal DNA. At the obtained phylogenetic tree, samples from the studied populations formed separate and clearly supported clades, that confirms their naturalness. ITS sequences of the analysed plants of each clade have group-specific substitutions, which mark their position on the phylogenetic tree. Analysis of both morphological traits and molecular-phylogenetic data allowed conducting species determination of plants from the genus Liparis. According to botanical literature, in the Amur region, two species were known: Liparis japonica and L. makinoana. The investigation of Liparis populations found in the field together with analysis results of morphological and molecular-phylogenetic data demonstrated that in the study area, besides L. japonica and L. makinoana, there are L. kumokiri and one more taxon, which could be a new species needed to be described in future. We clarified the distribution of these Liparis species in the study area by finding their new locations. The status of each population for studied species was appropriate at most of the locations. We noted the appropriate fruiting and offspring presence. At the studied location, permanent study plots could be established for conducting of further population-based studies. The obtained data on the status of plant populations will be used in the work on new editions of Red Data Books. In the study area, the presence of Liparis makinoana have not been confirmed by data of molecular-phylogenetic analysis.


molecular-phylogenetic study, morphological traits, phylogeny, population status, Protected Area, rare species, Red Data Book

Artice information

Received: 16.04.2020. Revised: 07.06.2020. Accepted: 29.06.2020.

The full text of the article

Cameron K.M. 2005. Leave it to the leaves: a molecular phylogenetic study of Malaxideae (Epidendroideae, Orchidaceae). American Journal of Botany 92(6): 1025–1032. DOI: 10.3732/ajb.92.6.1025
Chen S.C., Liu Z.J., Zhu G.H., Lang K.Y., Ji Z.H., Luo Y.B., Jin X.H., Cribb P.J., Wood J.J., Gale S.W., Ormerod P., Vermeulen J.J., Wood H.P., Clayton D., Bell A. 2009. Orchidaceae. In: Z.Y. Wu, P.H. Raven, D. Hong (Eds.). Flora of China. St. Louis: Science Press, Beijing et Missouri Botanical Garden Press. Vol. 25. P. 211–235.
Cheng T., Xu C., Lei L., Li C., Zhang Y., Zhou S. 2016. Barcoding the kingdom Plantae: New PCR primers for ITS regions of plants with improved universality and specificity. Molecular Ecology Resources 16(1): 138–149. DOI: 10.1111/1755-0998.12438
Darman G.F. 2015. Flora of the Muravyevskiy Natural Park. Scholarly Notes of Transbaikal State University 60(1): 11–16. [In Russian]
Dressler R.L. 1981. The orchids: natural history and classification. Cambridge, Massachusetts: Harvard University Press. 356 p.
Dressler R.L. 1993. Phylogeny and classification of the orchid family. Portland: Cambridge University Press. 314 p.
Edgar R.C. 2004. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research 32(5): 1792–1797. DOI: 10.1093/nar/gkh340
Efimov P.G. 2010. The genus Liparis (Orchidaceae) in Russia. Botanicheskii Zhurnal 95(10): 1458–1480. [In Russian]
Felsenstein J. 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39(4): 783–791. DOI: 10.1111/j.1558-5646.1985.tb00420.x
Gap S.O., Mi Y.C., Sun G.C., Myong G.C. 2001. Contrasting breeding systems Liparis kumokiri and L. makinoana (Orchidaceae). Annales Botanici Fennici 38(4): 281–284.
Hall T.A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium 41: 95–98.
Kim S.N., Kim Y.S. 1986. Morphological and cytological study on genus Liparis in Korea. Korean Journal of Plant Taxonomy 16(1): 59–88. DOI: 10.1007/s12374-010-9104-0
Kudrin S.G., Yakubov V.V. 2013. Illustrated Flora of the Khingan State Nature Reserve (Amur region): Vascular plants. Arkhara: Khingan State Nature Reserve. 335 p. [In Russian]
Lee C.S., Tsutsumi C., Yukawa T., Lee N.S. 2010. Two New Species of the Genus Liparis (Orchidaceae) from Korea Based on Morphological and Molecular Data. Journal of Plant Biology 53(3): 190–200. DOI: 10.1007/s12374-010-9104-0
Li L., Yan H. 2013. A remarkable new species of Liparis (Orchidaceae) from China and its phylogenetic implications. PLoS ONE 8(11): e78112. DOI: 10.1371/journal.pone.0078112
Maekawa F. 1971. The wild orchids of Japan in colour. Tokyo: Seibundo Shinkousha. 495 p. [In Japanese]
Miskina L.V. 1971. Landscape complexes of the Arkhara Lowland and south-western spurs of the Small Khingan and prospects for their economic development. PhD Thesis. Khabarovsk. 248 p. [In Russian]
Nevskiy S.A. 1935. Genus Liparis L. C. Rich. In: Flora of the USSR. Vol. 4. Leningrad: AS USSR. P. 601–604. [In Russian]
Ohwi J. 1965. Flora of Japan. Washington: Square Press. 1067 p.
Rabotnov T.A. 1950. Problems of investigating the composition of populations for the phytocoenology purpose. In: Problems of Botany 1: 465–483. [In Russian]
Red Data Book of the Amur region: Rare and Endangered Species of Animal, Plants and Fungi. Blagoveshchensk: Publishing House of the Far Eastern State Agrarian University, 2019. 501 p. [In Russian]
Red Data Book of the Russian Federation (plants and fungi). Moscow: KMK Scientific Press Ltd., 2008. 855 p. [In Russian]
Ronquist F., Teslenko M., van der Mark P., Ayres D.L., Darling A., Höhna S., Larget B., Liu L., Suchard M.A., Huelsenbeck J.P. 2012. MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61(3): 539–542. DOI: 10.1093/sysbio/sys029
Shibneva I.V. 2004. Liparis kumokiri (Orchidaceae) in the Russian Far East. Botanicheskii Zhurnal 89(10): 1633–1636. [In Russian]
Shibneva I.V. 2007. Notes on the Liparis (Orchidaceae) species from the Primorsky Krai. In: Plants in the monsoon climate. Vladivostok. P. 264–268. [In Russian]
Shibneva I.V. 2008. Species of the genus Liparis (Orchidaceae) in the south of the mainland part of the Russian Far East. In: Fundamental and applied problems of botany in early XXI century. Petrozavodsk. P. 148–150. [In Russian]
Shibneva I.V. 2011. What are Liparis japonica (Miq) Maxim. and L. makinoana Schltr. (Orchidaceae) – notes of the botanist. In: Protection and cultivation of orchids. St. Petersburg. P. 287–289. [In Russian]
Starchenko V.M., Darman G.F. 2002. Flora of the Botanical Garden of the Amur Scientific Centre FEB RAS. In: Introduction Centres of the Russian Far East, Research Results. Vladivostok: Dalnauka. P. 161–173. [In Russian]
Su Y.Y., Meng Y., Shi Y., Tang G.D., Liu Z.J. 2014. Liparis funingensis (Orchidaceae; Epidendroideae; Malaxidae), a new species from Yunnan, China: Evidence from morphology and DNA. Phytotaxa 166(1): 85–93. DOI: 10.11646/phytotaxa.166.1.6
Su Y.Y, Huang Y.L., Chen L.J., Zhang P.W., Liu Z.J., Zhang G.Q. 2015. Liparis wenshanensis, a new orchid species from China: Evidence from morphological and molecular analyses. Phytotaxa 204 (4): 253–264. DOI: 10.11646/phytotaxa.204.4.2
Swofford D.L. 2003. PAUP*. Phylogenetic Analysis Using Parsimony (*and Other Methods): Version 4. Massachusetts: Sinauer Associates. 142 p.
Tatarenko I.V. 1996. Orchids of Russia: life forms, biology, conservation issues. Moscow: Argus. 207 p. [In Russian]
Terentieva E.I., Degtjareva G.V., Efimov S.V., Samigullin T.Kh., Varlygina T.I. 2016. The use of morphological and molecular methods to determine the species affiliation of plants of the genus Liparis (Orchidaceae) in the Kedrovaya Pad State Nature Reserve. In: Problems of Botany in Southern Siberia and Mongolia. Barnaul: Publishing House of the Altai State University. P. 131–136. [In Russian]
Terentieva E.I., Degtjareva G.V., Efimov S.V., Samigullin T.Kh., Varlygina T.I. 2018. Taxonomic revision of the Liparis (Orchidaceae) species distributed in the Primorsky Krai and the Sakhalin region on the basis of the analysis of morphological and molecular data. In: Problems of Botany in Southern Siberia and Mongolia. Barnaul: Publishing House of the Altai State University. P. 440–444. [In Russian]
Terentieva E.I., Degtjareva G.V. Efimov S.V., Varlygina T.I. 2019. Species delimitation in Liparis (Orchidaceae) on the basis of the morphological and molecular data in the Muravyevskiy Park of the Sustainable Development. In: Problems of Botany in Southern Siberia and Mongolia. Barnaul: Publishing House of the Altai State University. P. 435–440. [In Russian]
Tsutsumi C. Yukawa T. 2008. Taxonomic status of Liparis japonica and L. makinoana (Orchidaceae); preliminary report. Bulletin of the National Museum of Nature and Science, Series B 34(2): 89–94.
Tsutsumi C., Yukawa T., Lee N.S., Lee C.S., Kato M. 2007. Phylogeny and comparative seed morphology of epiphytic and terrestrial species of Liparis (Orchidaceae) in Japan. Journal of Plant Research 120(3): 405–412. DOI: 10.1007/s10265-007-0077-0
Tsutsumi C., Yukawa T., Kato M. 2008a. Liparis purpureovittata (Orchidaceae) – a new species from Japan. Acta Phytotaxonomica et Geobotanica 59(1): 73–77. DOI: 10.18942/apg.KJ00004899890
Tsutsumi C., Yukawa T., Lee N.S., Lee C.S., Kato M. 2008b. A new species of Liparis from Japan and Korea. Acta Phytotaxonomica et Geobotanic 59(3): 211–218. DOI: 10.18942/apg.KJ00005124835
Vakhrameeva M.G., Varlygina T.I., Tatarenko I.V. 2014. Orchids of Russia: biology, ecology and conservation. Moscow: KMK Scientific Press Ltd. 437 p. [In Russian]
Valiejo-Roman C.M., Terentieva E.I., Samigullin T.H., Pimenov M.G. 2002. Relationships among genera in Saniculoideae and selected Apioideae (Umbelliferae) inferred from nrITS sequences. Taxon 51(1): 91–101. DOI: 10.2307/1554966
Varlygina T.I., Degtjareva G.V., Efimov S.V., Terentieva E.I. 2017. Population studies of the orchids in the Reserve Kedrovaya Pad. Biodiversity and Environment of Far East Reserves 1: 67–82. [In Russian]
Vyshin I.B. 1996. Orchids – Orchidaceae. In: Vascular plants of the Soviet Far East. St. Petersburg: Nauka. Vol. 8. P. 337–338. [In Russian]