Research paper Botanica Pacifica. A journal of plant science and conservation 2023. 12(2):155-162 Article first published online: 10 OCT 2023 | DOI: 10.17581/bp.2023.12209 Genetic diversity of the Rhododendron subsection Ledum species in Sakhalin Island: data from chloroplast DNA markers variability Diana R. Iunusova1 , Maria A. Polezhaeva1 & Elena A. Marchuk2 1 Institute of Plant and Animal Ecology UB RAS, Yekaterinburg, Russia 2 Botanical Institute-Garden FEB RAS, Vladivostok, Russia A high genetic variability was revealed in the populations of the Rhododendron subsection Ledum species complex in Sakhalin. In total, 94 individuals from natural populations of the four sympatric species of Labrador tea (R. tomentosum, R. subarcticum, R. tolmachevii and R. hypoleucum) were analyzed to evaluate the genetic variation in the territory of the Sakhalin Island and in two mainland populations. The haplotype diversity of five chloroplast intergenic spacers (trnH–psbA, petB–petD, 3’trnV (UAC)–ndhC, K2R–K707, atpB–rbcL) was quite high. There were 16 haplotypes (h1–h16) identified; h1, h2 and h4 are the most common, the rest were rare and no species-specific ones were found. Analysis of Molecular Variance (AMOVA) indicate that about 37 % of the genetic diversity is due to among population differences (FST = 0.37, p < 0.001). Molecular phylogenetic analysis based on chloroplast DNA data did not reveal a clear species structure, but revealed a trend towards the isolation of two genetic groups: including a group of samples from the southern and the middle parts of Sakhalin Island. Юнусова Д.Р., Полежаева М.А., Марчук Е.А. Генетическое разнообразие видов багульника на острове Сахалин: данные изменчивости маркеров хлоропластной ДНК. Изучено генетическое разнообразие 94 образцов из природных популяций четырех симпатричных видов багульников (подсекция Ledum рода Rhododendron: R. tomentosum, R. subarcticum, R. tolmachevii и R. hypoleucum) на территории о-ва Сахалин и в двух материковых популяциях. Изменчивость видов по пяти маркерным фрагментам хлоропластной ДНК (trnH–psbA, petB–petD, 3’trnV(UAC)–ndhC, K2R–K707, atpB–rbcL) оказалась довольно высокой. Выявлено 16 гаплотипов (h1-h16): h1, h2 и h4 – самые распространенные, остальные редкие, видоспецифичных не обнаружено. Анализ молекулярной дисперсии (AMOVA) показал, что около 37 % генетического разнообразия приходится на межпопуляционные различия (FST = 0,37, p < 0.001). Анализ родства, проведенный по данным хлоропластной ДНК, не выявил четкой видовой структуры, однако выявил тенденцию к обособлению двух генетических групп: включающей группу выборок из южной и средней частей о-ва Сахалин. Keywords: cpDNA haplotypes, Ericaceae, hybridization, Labrador tea, population structure, Russian Far East, species complex, гаплотипы хпДНК, Ericaceae, гибридизация, багульник, структура популяции, Дальний Восток России, видовой комплекс References Alix, K., P.R. Gérard, T. Schwarzacher & J.S. Heslop-Harrison 2017. Polyploidy and interspecific hybridization: partners for adaptation, speciation and evolution in plants. Annals of Botany 120(2):183-194. CrossRef Bandelt, H.J., P. Forster & A. Rohl 1999. Median-joining networks for inferring intraspecific phylogenies. Molecular Biology and Evolution 16:37-48. CrossRef Barkalov, V.Y. & A.A. Taran 2004. A checklist of vascular plants of the Sakhalin Island. In: Flora and Fauna of Sakhalin Island (Materials of International Sakhalin Island Project) (S.Yu. Storozhenko, ed.), pp. 39-66, Dal'nauka, Vladivostok (in Russian with English abstract). [Баркалов В.Ю., Таран А.А. 2004. Список видов сосудистых растений острова Сахалин // Растительный и животный мир острова Сахалин (Материалы Международного сахалинского проекта) / под ред. С.Ю. Стороженко. Владивосток: Дальнаука. С. 39-66]. Bruni, I., F. De Mattia, S. Fluch, C. Ferrari, M. Corazza, E. Dinelli & M. Labra 2016. Genetic introgression of hybrid Rhododendron × intermedium Tausch is habitat mediated: Evidences from south-eastern Alps (Italy). Plant Biosystems - An International Journal Dealing with All Aspects of Plant Biology 150(3):449-458. CrossRef Bush, E.A. & A.I. Poyarkova 1952. The family Ericaceae DC. In: Flora SSSR, vol. 18 (B.K. Shishkin & E.G. Bobrov, eds), pp. 22-93. Izd-vo AN SSSR, Moscow (in Russian). [Буш Е.А., Пояркова А.И. 1952. Семейство Вересковые - Ericaceae DC. Флора СССР. Т. 18 / под ред. Б.К. Шишкина и Е.Г. Боброва. Москва: Изд-во АН СССР. C. 22-93]. Cherepanov, S.K. 1995. Vascular plants of Russia and neighboring countries. Mir i sem'ya, Saint-Petersburg, 990 pp. (in Russian). [Черепанов С.К. 1991. Сосудистые растения России и сопредельных государств. Санкт-Петербург: Мир и семья. 990 с.]. Chiang, T.Y., B.A. Schaal & C.I. Peng 1998. Universal primers for amplification and sequencing a noncoding spacer between the atpB and rbcL genes of chloroplast DNA. Botanical Bulletin of Academia Sinica 39:245-250. Chung, J.D., T.P. Lin, Y.L. Chen, Y.P. Cheng & S.Y. Hwang 2007. Phylogeographic study reveals the origin and evolutionary history of a Rhododendron species complex in Taiwan. Molecular Phylogenetics and Evolution 42(1):14-24. CrossRef Dalgaard, V. & B. Fredskild 1993. Ledodendron vanhoeffeni (syn.: Rhododendron vanhoeffeni) refound in Greenland. Nordic Journal of Botany 13(3):253-255. CrossRef Dampc, A. & M. Luczkiewicz 2013. Rhododendron tomentosum (Ledum palustre). A review of traditional use based on current research. Fitoterapia 85:130-143. CrossRef Dampc, A. & M. Luczkiewicz 2015. Labrador tea - the aromatic beverage and spice: a review of origin, processing and safety. Journal of the Science of Food and Agriculture 95:1577-1583. CrossRef Devey, M.E., J.C. Bell, D.N. Smith, D.B. Neale & G.F. Moran 1996. A genetic linkage map for Pinus radiata based on RFLP, RAPD, and microsatellite markers. Theoretical and Applied Genetics 92:673-679. CrossRef Dumolin-Lapegue, S., M.-H. Pemonge & R.J. Petit 1997. An enlarged set of consensus primers for the study of organelle DNA in plants. Molecular Ecology 6:393-397. CrossRef Dupanloup, I., S. Schneider & L. Excoffier 2002. A simulated annealing approach to define the genetic structure of populations. Molecular Ecology 11:2571-2581. CrossRef Excoffier, L. & H. Lischer 2010. Arlequin suite ver 3.5: a new series of programs to perform population genetics analyses under Linux and Windows. Molecular Ecology Resources 10:564-567. CrossRef Goetsch, L., A.J. Eckert & B.D. Hall 2005. The molecular systematics of Rhododendron (Ericaceae): a phylogeny based upon RPB2 gene sequences. Systematic Botany 30(3):616-626. CrossRef Gurzenkov, N.N. 1973. Studies of chromosome numbers of plants from the south of the Soviet Far East. Komarov Memorial Lectures 20:47-61 (in Russian). [Гурзенков Н.Н. 1973. Исследование хромосомных чисел растений юга Дальнего Востока // Комаровские чтения. Вып. 20. С. 47-61]. Hall, T.A. 1999. Bioedit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41:95-98. Hamilton, M.B. 1999. Four primer pairs for the amplification of chloroplast intergenic regions with intraspecific variation. Molecular Ecology 8:521-523. Harmaja, H. 1990. 1990. New names and nomenclatural combinations in Rhododendron (Ericaceae). Annales Botanici Fennici 27(2):203-204. Hart, A., K. Kron & E. Gillespie 2017. Molecular phylogenetic analysis of the north-temperate Labrador Teas (Ericaceae: Rhododendron subsect. Ledum) suggests a complex genetic history. Journal of the Botanical Research Institute of Texas 11(1):53-65. CrossRef Khohryakov, A.P. & M.T. Mazurenko 1991. The family Ericaceae Juss. In: Vascular Plants of the Soviet Far East. Vol. 5 (S.S. Kharkevich, ed.), pp. 119-166, Nauka, Saint-Petersburg (in Russian). [Хохряков А.П., Мазуренко М.Т. 1991. Семейство 68. Вересковые - Ericaceae Juss. // Сосудистые растения советского Дальнего Востока / отв. ред. С.С. Харкевич. Санкт-Петербург: Наука. Т. 5. С. 119-166]. Kihlman, B. 2004. Hybrids between Ledums and lepidote Rhododendrons. Journal of the American Rhododendron Society 58:74-81. Komarov, V.L. 1932. Plant Identifier of the Far East Region. Leningrad, 622 pp. (in Russian). [Комаров В.Л. 1932. Определитель растений Дальневосточного края. Ленинград: Издательство Академии наук СССР. 622 с.]. Krestov, P.V., V.Yu. Barkalov & A.A. Taran 2004. Phytogeographical regionalization of the Sakhalin Island. In: Flora and Fauna of Sakhalin Island (Materials of International Sakhalin Island Project (S.Yu. Storozhenko, ed.), pp. 67-92, Dal'nauka, Vladivostok (in Russian with English abstract). [Крестов П.В., Баркалов В.Ю., Таран А.А. 2004. Ботанико-географическое районирование острова Сахалин // Растительный и животный мир острова Сахалин (Материалы Международного сахалинского проекта) / под ред. С.Ю. Стороженко. Владивосток: Дальнаука. С. 67-92]. Kron, K.A. & W.S. Judd 1990. Phylogenetic relationships within the Rhodoreae (Ericaceae) with specific comments on the placement of Ledum. Systematic Botany 15(1):57-68. CrossRef Krylov, P.N. 1937. Flora of Western Siberia. Izdanie Botanicheskoi Sektsii Tomskogo Obshchestva Ispytatelei Prirody, Tomsk, 331 pp. (in Russian). [Крылов П.Н. 1937. Флора Западной Сибири. Томск: Издание Ботанической Секции Томского Общества Испытателей Природы. 331 с.]. Kurashige, Y., J.I. Etoh, T. Handa, K. Takayanagi & T. Yukawa 2001. Sectional relationships in the genus Rhododendron (Ericaceae): Evidence from mat K and trn K intron sequences. Plant Systematics and Evolution 228:1-14. CrossRef Jiang, N., L. Man, W. Zhang, H.X. Dong, H.Y. Wang, M.R. Li, ... & M.Z. Sun 2016. Chloroplast view of the population genetics and phylogeography of a widely distributed shrub species, Rhododendron dauricum (Ericaceae). Systematic Botany 41(3):626-633. CrossRef Johnson, L.A. & D.E. Soltis 1995. Phylogenetic inference in Saxifragaceae sensu stricto and Gilia (Polemoniaceae) using matK sequences. Annals of the Missouri Botanical Garden 82:149-175. CrossRef Lantai, K. & B. Kihlman. 1995. The chromosome number of Ledum palustre ssp. decumbens and of some related taxa. Hereditas 122(2):181-184. CrossRef Löhne, C. & T. Borsch 2005. Molecular evolution and phylogenetic utility of the petD group II intron: a case study in basal angiosperms. Molecular Biology and Evolution 22(2):317-332. CrossRef Manen, J.F., A. Natali & F. Ehrendorfer 1994. Phylogeny of Rubiaceae-Rubieae inferred from the sequence of a cpDNA intergene region. Plant Systematics and Evolution 190:195-211. CrossRef Matthews, J.V. 1979. Tertiary and Quaternary environments: historical background for an analysis of the Canadian insect fauna. The Memoirs of the Entomological Society of Canada 111(108):31-86. CrossRef Mazurenko, M.T. & L.P. Khokhryakov 1987. Evolution of biomorphs and speciation in Far Eastern ledums. Zhurnal Obshchei Biologii 48(2):213-221 (in Russian). [Мазуренко М.Т., Хохряков Л.П. 1987. Эволюция биоморф и таксонообразование у дальневосточных багульников // Журнал общей биологии. Т. 48, № 2. С. 213-221]. Mĕsíček, J. & V. Javůrková-Jarolímová 1992. List of Chromosome Numbers of the Czech Vascular Plants. Academia, Praha, 144 pp. Nei, M. 1972. Genetic distance between populations. American Naturalist 106:283-292. CrossRef Rambaut, A. & A. Drummond 2009. FigTree v1.3.1: Tree figure drawing tool. Institute of Evolutionary Biology, Edinburgh, UK. Pietsch, T.W., V.V. Bogatov, S.Yu. Storozhenko, A.S. Lelej, V.Yu. Barkalov, H. Takahashi, S.L. Joneson, S.K. Kholin, K.A. Glew, J.A. Harpel, ... & D.E. Stevenson 2012. Biodiversity and biogeography of Sakhalin Island. In: Flora and fauna of North-West Pacific islands (Materials of International Kuril Island and International Sakhalin Island Projects), (S.Yu. Storozhenko, ed.), pp. 11-78, Dal'nauka, Vladivostok. Pletnev, S.P. 2004. Geological development of Sakhalin Island. In: Flora and Fauna of Sakhalin Island (Materials of International Sakhalin Island Project), (S.Yu. Storozhenko, ed.), pp. 11-22, Dal'nauka, Vladivostok (in Russian with English abstract). [Плетнев С.П. 2004. Историко-геологическое развитие острова Сахалин // Растительный и животный мир острова Сахалин (Материалы Международного сахалинского проекта) / под ред. С.Ю. Стороженко. Владивосток: Дальнаука. С. 11-22]. Polezhaeva, M.A., E.A. Pimenova, N.A. Tikhonova & O.S. Korchagina 2018. Plastid DNA diversity and genetic divergence within Rhododendron dauricum s.l. (R. dauricum s.s., R. ledebourii, R. sichotense and R. mucronulatum; Ericaceae). Plant Systematics and Evolution 304:763-774. CrossRef Polezhaeva, M.A., D.R. Iunusova, N.A. Tikhonova, A.N. Polezhaev & M.N. Koldaeva 2022. Genetic identification of closely related endangered Rhododendron species from East Asia. Russian Journal of Genetics 58(1):116-121. CrossRef Probatova, N.S., V.Yu. Barkalov & E.G. Rudyka 2007. Karyology of the flora of Sakhalin and Kuril Islands. Dal'nauka, Vladivostok, 392 pp. (in Russian). [Пробатова Н.С., Баркалов В.Ю., Рудыка Э.Г. 2007. Кариология флоры Сахалина и Курильских остров. Владивосток: Дальнаука. 392 с.]. Ronquist, F. & J.P. Huelsenbeck 2003. MrBAYES 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19(12):1572-1574. CrossRef Shaw, J., E.B. Lickey, E.E. Schilling & R.L. Small 2007. Comparison of whole chloroplast genome sequences to choose noncoding regions for phylogenetic studies in angiosperms: the tortoise and the hare III. American Journal of Botany 94(3):275-288. CrossRef Sugawara, Sh. 1940. Illustrated flora of Saghalien with descriptions and figures of phanerogams and higher cryptogams indigenous to Saghalien. Vol. 4 vol. 4: Diapensiacae-Asteraceae, Tokyo, pp. 1439-1957. Taberlet, P.T., L. Geilly, G. Patou & J. Bouvet 1991. Universal primers for amplification of three noncoding regions of chloroplast DNA. Plant Molecular Biology 17:1105-1109. CrossRef Theqvist, K. 2011. Ploidy level of Ledum species. http://rosebayblog.blogspot.com/search/label/ledum Last accessed 05.04.2023. Theqvist, K. 2013. Hybridity of lepidotes in Rhododendron subsection Ledum based on some hybridization results. https://www.rhodogarden.com/cross/hybridity.html Last accessed 06.04.2023. Tolmachev, A.I. 1953. Toward the cognition of Eurasian species of the genus Ledum L. Botanicheskie Materialy Gerbariya BIN AN SSSR 15:197-207 (in Russian). [Толмачев А.И. 1953. К познанию евразиатских видов рода Ledum L. // Ботанические материалы гербария БИН АН СССР. Т. 15. С. 197-207]. Tolmachev, A.I. 1974. Guide to the higher plants of Sakhalin and the Kuril Islands. Nauka, Leningrad, 372 pp. (in Russian). [Толмачев А.И. 1974. Определитель высших растений Сахалина и Курильских островов. Ленинград: Наука. 372 с.]. Voroshilov, V.N. 1982. Guide to the plants of the Soviet Far East. Nauka, Moscow, 674 pp. (in Russian). [Ворошилов В.Н. 1982. Определитель растений советского Дальнего Востока. Москва: Наука. 674 с.]. Zha, H.G., R.I. Milne & H. Sun 2008. Morphological and molecular evidence of natural hybridization between two distantly related Rhododendron species from the Sino-Himalaya. Botanical Journal of the Linnean Society 156(1):119-129. CrossRef
|