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0008-7114 2020 3 73 – n. Vol. Caryologia 2020 International Journal of Cytology, Vol. 73 - n. 3 Cytosystematics and Cytogenetics Caryologia International Journal of Cytology, Cytosystematics and Cytogenetics International Journal of Cytology, FIRENZE PRESSUNIVERSITY Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics Caryologia is devoted to the publication of original papers, and occasionally of reviews, about plant, animal and human kar- yological, cytological, cytogenetic, embryological and ultrastructural studies. Articles about the structure, the organization and the biological events relating to DNA and chromatin organization in eukaryotic cells are considered. Caryologia has a strong tradition in plant and animal cytosystematics and in cytotoxicology. Bioinformatics articles may be considered, but only if they have an emphasis on the relationship between the nucleus and cytoplasm and/or the structural organization of the eukaryotic cell. Editor in Chief Associate Editors Alessio Papini Alfonso Carabez-Trejo - Mexico City, Mexico Dipartimento di Biologia Vegetale Katsuhiko Kondo - Hagishi-Hiroshima, Japan Università degli Studi di Firenze Canio G. Vosa - Pisa, Italy Via La Pira, 4 – 0121 Firenze, Italy Subject Editors Mycology Plant Cytogenetics Histology and Cell Biology Renato Benesperi Lorenzo Peruzzi Alessio Papini Università di Firenze, Italy Università di Pisa Università di Firenze Human and Animal Cytogenetics Plant Karyology and Phylogeny Zoology Michael Schmid Andrea Coppi Mauro Mandrioli University of Würzburg, Germany Università di Firenze Università di Modena e Reggio Emilia Editorial Assistant Sara Falsini Università degli Studi di Firenze, Italy Editorial Advisory Board G. Berta - Alessandria, Italy G. Delfino - Firenze, Italy M. Mandrioli - Modena, Italy D. Bizzaro - Ancona, Italy S. D'Emerico - Bari, Italy G. C. Manicardi - Modena, Italy A. Brito Da Cunha - Sao Paulo, Brazil F. Garbari - Pisa, Italy P. Marchi - Roma, Italy E. Capanna - Roma, Italy C. Giuliani - Milano, Italy M. Ruffini Castiglione - Pisa, Italy D. Cavalieri - San Michele all'Adige, Italy M. Guerra - Recife, Brazil L. Sanità di Toppi - Parma, Italy E. H. Y. Chu - Ann Arbor, USA W. Heneen - Svalöf, Sweden C. Steinlein - Würzburg, Germany R. Cremonini - Pisa, Italy L. Iannuzzi - Napoli, Italy J. Vallès - Barcelona, Catalonia, Spain M. Cresti - Siena, Italy J. Limon - Gdansk, Poland Q. Yang - Beijing, China G. Cristofolini - Bologna, Italy J. Liu - Lanzhou, China P. Cro sti - Milano, Italy N. Mandahl - Lund, Sweden Cover: figure from the article inside by Rodríguez-Domínguez et al. "Physical mapping of 45S and 5S rDNA in two Sprekelia formosissima cyto- types (Amaryllidaceae) through Fluorescent In Situ Hybridization (FISH)". Karyogram of diploid Sprekelia formosissima. Caryologia International Journal of Cytology, Cytosystematics and Cytogenetics Volume 73, Issue 3 - 2020 Firenze University Press Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics Published by Firenze University Press – University of Florence, Italy Via Cittadella, 7 - 50144 Florence - Italy http://www.fupress.com/caryologia Copyright © 2020 Authors. The authors retain all rights to the original work without any restrictions. Open Access. This issue is distributed under the terms of the Creative Commons Attribution 4.0 International License (CC-BY-4.0) which permits unrestricted use, distribution, and reproduction in any medium, provided you give ap- propriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication (CC0 1.0) waiver applies to the data made available in this issue, unless otherwise stated. Firenze University Press Caryologia www.fupress.com/caryologia International Journal of Cytology, Cytosystematics and Cytogenetics Species delimitation in the genus Cousinia Cass. (Family Asteraceae), sections Citation: N. Atazadeh, M. Sheidai, F. Attar, F. Koohdar (2020) Species delimita- Cynaroideae Bunge and Platyacanthae Rech. f.: tion in the genus Cousinia Cass. (Fam- ily Asteraceae), sections Cynaroideae morphometry and molecular analysis Bunge and Platyacanthae Rech. f.: morphometry and molecular analysis. Caryologia 73(3): 3-12. doi: 10.13128/ caryologia-275 Neda Atazadeh1,*, Masoud Sheidai1, Farideh Attar2, Fahimeh Kooh- dar1 Received: May 30, 2019 1 Faculty of Life Sciences & Biotechnology, Shahid Beheshti University, Evin, Tehran, Iran Accepted: April 05, 2020 2 Central Herbarium of Tehran University, School of Biology, College of Science, Univer- Published: December 31, 2020 sity of Tehran, P.O. Box: 14155, Tehran, Iran * Corresponding author: Email: [email protected] Copyright: © 2020 N. Atazadeh, M. Sheidai, F. Attar, F. Koohdar. This is an open access, peer-reviewed article Abstract. The genus Cousinia of the tribe Cardueae with about 700 species is one of published by Firenze University Press the most diverse genera in Central and southwest Asia. The section Cynaroides with (http://www.fupress.com/caryologia) 89 species is the largest section of the genus. Due to the controversy in the number of and distributed under the terms of the Cousinia species and their delineation, the first step in studying the genus is to iden- Creative Commons Attribution License, tify and delimit presumed species. Species delimitation is usually difficult in the spe- which permits unrestricted use, distri- cies with overlaps in their morphological features. Therefore, we used a combination bution, and reproduction in any medi- of morphological and molecular markers (ISSRs) to carry out delimitation in 204 taxa um, provided the original author and source are credited. of 68 Cousinia species whitin the Cynaroideae and Platyacanthae sections. The spe- cies delineation based on morphometry and ISSR data were done by UPGMA cluster- Data Availability Statement: All rel- ing. The samples of each species were placed close to each other and formed a single evant data are within the paper and its sub-cluster, separated from the other studied Cousinia species. In the present study Supporting Information files. the studied Cousinia species within Cynaroideae and Platyacanthae sections could be delimited from each other based on ISSR and morphological data. Therefore, using Competing Interests: The Author(s) declare(s) no conflict of interest. ISSR and morphological data can be useful in identifying and delineating crucial spe- cies. The Mantel test performed between morphological distance and Nei genetic dis- tance produced non-significant correlation. This result also supports distance analyses of the trees and reveals that the two dendrograms are not correlated. Some possible reasons for this incongruence are proposed: the high number of taxa in the genus Cousinia, morphological traits homoplasious, convergent evolution and incomplete lin- eage sorting. Keywords: Cousinia, Cynaroideae, ISSR, morphometry. INTRODUCTION The genus Cousinia Cass. of the tribe Cardueae (Family Asteraceae) with about 700 species is one of the most diverse genera in Central and southwest (SW) Asia After Senecio L. (c. 1500 species) and Vernonia Schreb. (c. 1000 species) (Tscherneva 1962; Rechinger 1972, 1979; Frodin 2004; Attar and Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics 73(3): 3-12, 2020 ISSN 0008-7114 (print) | ISSN 2165-5391 (online) | DOI: 10.13128/caryologia-275 4 Neda Atazadeh, Masoud Sheidai, Farideh Attar, Fahimeh Koohdar Ghahreman 2006; Susanna and Garcia-Jacas 2006; Attar Due to the controversy in the number of Cousinia and Djavadi 2010; Mehregan and Assadi 2016; Minaeifar species and their delineation, the first step in studying et al. 2016; Rastegar et al. 2017, 2018). Due to the exten- the genus is to identify and delimit presumed species. sive morphological variability in the genus, Cousinia Species delimitation is usually difficult in the species taxonomy is complicated and controversial (Mabber- with overlaps in their morphological features (Wiens ley 1990; Haffner 2000; Susanna et al. 2003). The genus 2007). In such cases, combined morphological and Cousinia contains more than 400 species in SW Asia, molecular data have been used to delimit these taxonom- with the highest number of species in the Flora Iranica ic identities (Duminil and Di Michele 2009; Minaeifar et area, out of which 379 are endemic. These species are al. 2016; Hassanpour et al. 2018; Eftekharian et al. 2018). distributed in mountainous regions of Iran, Afghani- Different molecular markers have been used in plant stan and Turkmenistan (Rechinger 1986; Knapp 1987). taxonomy and phylogeny, but some of them such as inter- Although the exact number of Cousinia species in Iran simple sequence repeats (ISSRs) seems to be very effi- is still unknown, about 270 species have been reported cient in delineating species, varieties, ecotype and even till now (Assadi 2009; Attar and Djavadi 2010). Out of genotypes of a single species (See for example, Sheidai et these, nearly 200 endemic Cousinia species occur in Iran al. 2012, 2013; Safaei et al. 2016; Eftekharian et al. 2018). (Djavadi et al. 2007; Zare et al. 2013). The Cousinia spe- Therefore, we used a combination of morphological and cies are distributed in 70 sections (Rechinger 1986). The molecular markers (ISSRs) to carry out Cousinia species section Cynaroideae Bunge with 89 species is the larg- delimitation in sections Cynaroideae and Platyacanthae. est