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Genome size and chromosome number in Echinops (Asteraceae, Cardueae) in the Aegean and Balkan regions: technical aspects of nuclear DNA amount assessment and genome evolution in a phylogenetic frame

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Abstract

This work focuses on the representatives of genus Echinops (Asteraceae, Cardueae) in the Aegean and Balkan regions, from the perspective of their genome evolution. Chromosome numbers were determined by orcein staining in 14 populations of nine taxa, and DNA contents were assessed by flow cytometry in 24 populations of nine taxa. A molecular phylogeny based on the internal transcribed spacer (ITS) and trnL-trnF and including first sequences for two taxa (Echinops sphaerocephalus subsp. taygeteus and E. spinosissimus subsp. neumayeri) provided a framework for discussing genome changes. From a methodological point of view, similar C-DNA value estimates were obtained when measuring, for a same population, fresh leaves from adult plants collected in the field and from cultivated seedlings. Conversely, despite giving the appearance of being correct (e.g., low coefficient of variation), genome size assessed using silica gel-preserved material differs significantly from values obtained for the same populations with fresh material. Nevertheless, silica gel-preserved material may still provide rough estimates of genome size for, e.g., inferring ploidy level. Suitable—non-silica gel-based—DNA amounts assessed for 23 populations range from 2C = 6.52 pg (E. spinosissimus subsp. neumayeri) to 2C = 9.37 pg (E. bannaticus). Chromosome counts were established for the first time for Echinops graecus (2n = 32), E. sphaerocephalus subsp. albidus (2n = 32), E. sphaerocephalus subsp. taygeteus (2n = ca. 30), and E. spinosissimus subsp. neumayeri (2n = 28). Genome size and chromosome number are confirmed as crucial parameters for deciphering lineage diversification within the genus Echinops.

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Acknowledgments

We thank Dr. K. Tan, Dr. J. Vigo, and Dr. F. Pustahija for their help in collecting material, M. Veny for taking care of the living plants, M. Mumbrú, Dr. J. Comas, and Dr. R. Álvarez for their support in the flow cytometry assessments, and an anonymous reviewer for useful comments. Alastair Plant is also acknowledged for the revision of the English in the manuscript. This work was supported by the MICINN of the Spanish Government (projects CGL2007-64839-C02-01, CGL2007-64839-C02-02, CGL2010-22234-C02-01/BOS, and CGL2010-22234-C02-02/BOS) and the GReB (Grup de Recerca en Biodiversitat i Biosistemàtica Vegetals; Generalitat de Catalunya, project 2009SGR439). I.S.-J. received a FPU grant from the Ministerio de Educación and O.H. a Juan de la Cierva postdoctoral grant from the Ministerio de Ciencia e Innovación, Spain.

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Authors and Affiliations

  1. Institut Botànic de Barcelona (IBB-CSIC-ICUB), Passeig del Migdia s.n., 08038, Barcelona, Catalonia, Spain

    Ismael Sánchez-Jiménez & Teresa Garnatje

  2. Department of Environmental and Plant Biology, Plant Development and Evolution, Ohio University, 500 Porter Hall, Athens, OH, 45701, USA

    Oriane Hidalgo

  3. Laboratori de Botànica, Facultat de Farmàcia, Universitat de Barcelona, Av. Joan XXIII s.n., 08028, Barcelona, Catalonia, Spain

    Oriane Hidalgo & Joan Vallès

  4. Managerial Decision Sciences, IESE Business School, Universidad de Navarra, Av. Pearson 21, 08032, Barcelona, Catalonia, Spain

    Miguel Ángel Canela

  5. Unité Ecologie, Systématique and Evolution, UMR CNRS 8079, AgroParisTech, Université Paris-Sud, Bât. 360, 91405, Orsay Cedex, France

    Sonja Siljak-Yakovlev

  6. Institute “Mountain and Sea”, Franjevački put 1, 21300, Makarska, Croatia

    Marija Edita Šolić

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  1. Ismael Sánchez-Jiménez
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Correspondence to Ismael Sánchez-Jiménez.

Appendix

Appendix

Taxa and GenBank accession numbers [ITS; trnL-trnF] of sequences included in the phylogenetic analysis. Data of sequences firstly published in this article are written in bold; the origin of remaining material is indicated in Sánchez-Jiménez et al. (2010).

Echinops albicaulis Kar. & Kir. [AY538638; GU134528]; E. angustilobus S.Moore [GU116505; GU134531]; E. arachniolepis Rech.f. [GU116486; GU134532]; E. bannaticus Rochel ex Schrad. [JQ291287; JQ291297]; E. cephalotes DC. [GU116487; GU134535]; E. ceratophorus Boiss. [GU116488; GU134536]; E. chantavicus Trautv. [GU116489; GU134537]; E. chardinii Boiss. & Buhse [GU116490; GU134538]; E. chloroleucus Rech.f. [GU116491; GU134539]; E. cyanocephalus Boiss. & Hausskn. [GU116492; GU134540]; E. dahuricus Fisch. [GU116493; GU134541]; E. dasyanthus Regel & Schmalh. [GU116494; GU134542]; E. dichrous Boiss. & Hausskn. [GU116495; GU134543]; E. dubjanskyi Iljin [GU116496; GU134544]; E. ecbatanus Bornm. ex Rech.f. [(AF319073, AF319127); GU134545]; E. echinatus Roxb. [GU116497; GU134546]; E. elbursensis Rech.f. [GU116498; GU134547]; E. elymaiticus Born [GU116499; GU134548]; E. emiliae Schwarz ex P.H.Davis [AY538641; GU134549]; E. endotrichus Rech.f. [GU116500; GU134550]; E. exaltatus Schrad. [GU116501; GU134551]; E. fastigiatus Kamelin & Tscherneva [GU116502; GU134552]; E. fontqueri Pau [AY538632; GU134554]; E. freitagii Rech.f. [GU116504; GU134555]; E. gaillardotii Boiss. [GU116507; GU134557]; E. ghoranus Rech.f. [GU116508; GU134558]; E. glaberrimus DC. [GU116509; GU134559]; E. gmelinii Turcz. [GU116510; GU134560]; E. graecus Mill. [JQ291289; JQ291299]; E. griffithianus Boiss. [GU116512; GU134562]; E. hedgei Kit Tan [AY538648; GU134563]; E. heteromorphus Bunge [AY538630; GU134564]; E. hoehnelii Schweinf. [GU116506; GU134565]; E. hololeucus Rech.f. [GU116513; GU134567]; E. humilis M.Bieb [GU116514; GU134568]; E. hystrichoides Kit Tan [GU116515; GU134570]; E. ilicifolius Bunge [GU116516; GU134571]; E. integrifolius Kar. & Kir. [GU116517; GU134572]; E. cf. karatavicus [AY538633; AY772325]; E. karatavicus Regel & Schmalh. [GU116518; GU134573]; E. knorringianus Iljin [GU116519; GU134574]; E. kotschyi Boiss. [GU116520; GU134575]; E. latifolius Tausch. [GU116521; GU134576]; E. leucographus Bunge [GU116522; GU134577]; E. lipskyi Iljin [GU116523; GU134578]; E. longifolius A.Rich [GU116524; GU134579]; E. maracandicus Bunge [GU116526; GU134581]; E. macrocephalus Sibth. & Sm. [GU116543; GU134584]; E. nanus Bunge [GU116528; GU134585]; E. nitens Bornm. [GU116529; GU134586]; E. nizvanus Rech.f. [GU116530; GU134587]; E. nuratavicus A.D.Li [GU116531; GU134588]; E. obliquilobus Iljin [AY538640; GU134602]; E. onopordum P.H.Davis [AY538642; GU134589]; E. orientalis Trautv. [GU116532; GU134590]; E. ossicus K.Koch [AY538644; GU134556]; E. parviflorus Boiss. & Buhse [GU116533; GU134591]; E. persicus Steven ex Fisch. [AY538639; AY772324]; E. platylepis Trautv. [AY538635; GU134569]; E. polygamus Bunge [GU116534; GU134593]; E. przewalskyi Iljin (LE) [GU116535; GU134594]; E. przewalskyi Iljin (XINA41JV) [GU116537; GU134596]; E. pungens Trautv. [GU116538; GU134597]; E. ritro L. (SJ9) [JQ291292; JQ291302]; E. ritro L. (SJ45) [JQ291291; JQ291301]; E. ritro subsp. meyeri (DC.) Kožuharov [GU116527; GU134582]; E. ritro L. subsp. ritro [GU116544; GU134599]; E. ritro L. subsp. siculus (Strobl) Greuter [AY538652; GU134604]; E. ritrodes Bunge [GU116539; GU134598]; E. setifer Iljin [GU116540; GU134603]; E. sphaerocephalus L. subsp. albidus (Boiss. & Spruner) Kožuharov [JQ291286; JQ291296]; E. sphaerocephalus L. subsp. sphaerocephalus [JQ291293; JQ291303]; E. sphaerocephalus L. subsp. taygeteus (Boiss. & Heldr.) Kožuharov [JQ291295; JQ291305]; E. spiniger Iljin [GU116542; GU134607]; E. spinosissimus Turra [AY826283; AY772326]; E. spinosissimus subsp. bithynicus (Boiss.) Greuter [JQ291288; JQ291298]; E. spinosissimus Turra subsp. macrolepis (Boiss.) Greuter [GU116547; GU134615]; E. spinosissimus Turra subsp. neumayeri (Vis.) Kožuharov [JQ291290; JQ291300]; E. spinosissimus Turra subsp. spinosissimus [JQ291294; JQ291304]; E. strigosus L. [AY538653; GU134528]; E. szovitsii DC. [AY538650; GU134610]; E. talassicus Golosk. [AY538631; GU134611]; E. tjanschanicus Bobrov [GU116545; GU134612]; E. tournefortii Ledeb. [AY538646; GU134613]; E. transcaucasicus Iljin [GU116546; GU134614].

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Sánchez-Jiménez, I., Hidalgo, O., Canela, M.Á. et al. Genome size and chromosome number in Echinops (Asteraceae, Cardueae) in the Aegean and Balkan regions: technical aspects of nuclear DNA amount assessment and genome evolution in a phylogenetic frame. Plant Syst Evol 298, 1085–1099 (2012). https://doi.org/10.1007/s00606-012-0618-4

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