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Masarykova Univerzita Přírodovědecká Fakulta MASARYKOVA UNIVERZITA PŘÍRODOVĚDECKÁ FAKULTA ÚSTAV BOTANIKY A ZOOLOGIE Význam polyploidie, hybridizácie a asexuálneho rozmnožovania v evolúcii cievnatých rastlín Habilitačná práca Marly, január 2012 Patrik Mráz 2 Obsah Poďakovanie………………………………………………..……………………….…….…...5 Abstrakt……………………….…………………………………………………….…….……7 1. ZOZNAM PUBLIKÁCIÍ – PRÍLOH UCHÁDZAČA ZARADENÝCH DO HABILITAČNEJ PRÁCE………..…………………………………...…………..……..9 2. ÚVOD……………………………...………………………………………………...…….15 3. POLYPLOIDIA……………………………………...…………………………….………16 3.1. Význam polyploidie pre tvorbu reprodukčných bariér…………………………....……16 3.2. Význam polyploidie z hľadiska tvorby nových fenotypov..……………………………17 3.3. Typ polyploidie a jeho rozlíšenie…………………………………………….…..…….18 3.4. Polytopický vznik polyploidov a geografické paterny…………………………...…….19 3.5. Frekvencia polyploidie………………………………………………………………....20 3.6. Mechanizmy vzniku polyploidov………………………………………………………21 3.7. Mechanizmy koexistencie polyploidov…………………………………………..…….22 4. HYBRIDIZÁCIA……………………………………………………………………….….23 5. APOMIXIA…………………………………………………………………………….…..25 6. ZÁVER………...……………………………………………………………….………….28 7. LITERATÚRA…………………………………………………………………….……….28 3 4 Poďakovanie Hoci sa rodičom a blízkym zvykne ďakovať až na konci tejto kapitoly ja začnem práve nimi, nakoľko to bol môj otec, ktorý vo mne vzbudil záujem o prírodu. Spolu s bratom nás každodenne brával na prechádzky prakovskými lesmi, dolinami a dolkami, kde sme pozorovali všetko, čo sa hýbalo aj nehýbalo, a to bez ohľadu na prítomnosť či neprítomnosť chlorofylu v bunkách. Za to, že tento záujem pretrval podnes, a ktorý sa mi stal aj životným povolaním ďakujem aj mamine, manželke Vierke a mojim deťom. Tí všetci ma podporovali v mojom bádateľskom úsilí a vytvárali nevyhnutné zázemie potrebné pre normálny život. Im patrí moja najväčšia vďaka. Za to, že som šiel študovať biológiu, môže v dobrom Mária Kötelešová, moja triedna profesorka a učiteľka biológie z gelnického gymnázia. Na vysokej škole mi v profesionálnej orientácii pomohol Pavol Mártonfi, ktorý mi od počiatku nechal voľnú ruku bádať, a ktorý mi sprostredkoval kontakt s Karolom Marholdom. Karolovi, ale aj ďalším spolupracovníkom z Botanického ústavu SAV vďačím za veľa. Moje neskoršie vedecké smerovanie do výraznej miery ovplyvnili aj krátkodobé návštevy v Prahe, či dlhodobé pobyty na pracoviskách v Grenobli a vo Fribourgu. Stretnutia, spolupráca a diskusie s mnohými kolegami z týchto pracovísk boli nesmierne prínosné. Osobitne by som chcel spomenúť aspoň niektorých kolegov a priateľov. Jindra Chrtek má najväčšiu zásluhu na tom, že ma nenásilne presvedčil, že asexuálne jastrabníky sa oplatí študovať, aj keď v počiatkoch som z nich mával často hlavybôľ. Spolupráca s Jindrom bola a je nesmierne plodná, podobne tak spolupráca s mojou bývalou študentkou Bašou Šingliarovou. Phillip Choler a Heinz Müller-Schärer formovali moje ‘ekologické povedomie’. Heinzovi vďačím aj za ponuku pracovať na veľmi zaujímavom a z hľadiska inváznej biológie modelovom druhu – nevädzi porýnskej, a takisto ďalším kolegom z Fribourgu za veľmi podnetné prostredie. V laboratóriu a na experimentálnom políčku či v skleníku mi pomáhali Milka Sasáková, Delphine Rioux a Anne-Catherine Cossy. Zvlášť stimulujúca a obohacujúca bola práca so študentami (abecedne Alexandre Farkas, David Frey, Emilie Gex-Fabry, Katarína Kovalčiková, Karol Krak, Sandrine Michel, Juraj Paule, Valentine Renevey, Timea Rezešová, Světlana Sychrová, Barbora Šingliarová, Elham Tarbush, Daniela Tomčíková). Vďaka! 5 Táto práca vznikla spojením 19 publikovaných a jednej do redakcie zaslanej a predbežne do tlače prijatej štúdie, na ktorých sa podieľalo 38 spoluautorov. Bez nich by som nemohol dosiahnuť tu prezentované výsledky. Im všetkým ďakujem. Zároveň ďakujem redakcii Annales Botanici Fennici za umožnenie zaradiť pdf článku uverejnenon v tomto časopise (Príloha G) do predkladanej habilitačnej práce. 6 Abstract Interspecific hybridization and polyploidization and, in a lesser extent, apomixis (asexual seed reproduction) have played a substantial role in the evolution of vascular plants. Although all three processes are often closely associated, their contribution to speciation is different. Using examples from three genera of sunflower family (Centaurea, Hieracium and Pilosella), we briefly summarize in this thesis the different aspects of these evolutionary mechanisms, which have been studied by the author and his collaborators over the last ten years. Interspecific hybridization results in phenotypic and genotypic alterations, and if such an ‘evolutionary novelty’ is reproductively isolated from parental taxa and its reproduction is assured then a new hybridogeneous species can arise. We provide molecular evidence for hybrid origin and morphological differentiation of the allotetraploid Centaurea stoebe and the allopolyploid Pilosella alpicola s. str. (Appendices B, C, M, N). These hybridogeneous species were reproductively isolated from their ancestors either by a ploidy barrier (C. stoebe) and / or by apomictic reproduction (P. alpicola s.str.) (Appendices A, C). We hypothesize that interspecific hybridization has triggered an important shift from a monocarpic life cycle in the diploid C. stoebe to a polycarpic perennial life cycle in tetraploid cytotype. This in turn might explain the colonization success of a tetraploid cytotype in North America, where it has become invasive. In contrast, autopolyploidization in Pilosella rhodopea did not lead to morphological nor genetic differentiaton, probably due to very effective and intensive intercytotype gene flow (Appendix D). Our results show that the triploid cytotype of P. rhodopea creates an important intercytotype bridge, forming tetraploids through frequent unreduced gamete formation (Appendix C). Furthemore, cytogeographic and genetic data suggest multiple origins of autopolyploidy and a primary diploid-polyploid contact zone in P. rhodopea. The primary contact zones where new polyploids arise from resident diploids have only rarely been reported in vascular plants (Apendices C, D). The intercytotype interactions were also studied in five mixed-ploidy populations of Centaurea stoebe. By combining different methodological approaches, we found strong microspatial and microhabitat segregation of cytotypes in every site. Such segregation was driven by anthropogenic disturbance and later immigrations of tetraploids into already established diploid populations. Our results thus highlight the importance of non-adaptive spatio-temporal processes in explaining microhabitat and microspatial segregation of cytotypes. (Appendix A). 7 The issue of natural hybridization in the genus Hieracium, its frequency and direction, was addressed in three studies (Appendices L, O, P). They provided the first convincing evidence on recent, though rare, homoploid hybridization in the genus. In addition to natural hybridization, we also performed a pionier experimental hybridization in the same genus. The most important result was a discovery of induced autogamy in Hieracium – a phenomenom which might play an important role in reproductive isolation of otherwise strictly self- incompatible taxa (Appendices Q, R, S). Using karyological and flow-cytometric methods, we assessed new or corroborated previous chromosome counts / ploidy levels for many Pilosella and Hieracium taxa (Appendices C, E, F, G, H, I, J, K). In addition, for many hawkweed species we have determined the mode of reproduction by performing either a castration experiment or flow- cytometric seed screening analyses (Appendices C, G, I, Q). Interestingly, we have found a correlation between hybridization and apomictic reproduction in the allopolyploid P. alpicola s.str. but not in the autopolyploid P. rhodopea that exclusively reproduces sexually (Appendix C). These results thus stress the importance of polyploidy and apomictic reproduction as the stabilization mechanisms in the evolution of Hieracium and Pilosella. 8 1. ZOZNAM PUBLIKÁCIÍ – PRÍLOH UCHÁDZAČA ZARADENÝCH DO HABILITAČNEJ PRÁCE A. Mráz P, Španiel S, Keller A, Bowmann G, Farkas A, Šingliarová B, Rohr RP, Broennimann O, Müller-Schärer H. Anthropogenic disturbance as a driver of microspatial and microhabitat segregation of cytotypes in diploid-tetraploid contact zones. (rukopis pripravený na odoslanie). Autorský podiel uchádzača: 80%. PM navrhol a koordionoval štúdiu, podieľal sa na zbere rastlinného materiálu, na meraniach rastlín v teréne, spolupodieľal sa na cytometrických a karyologických analýzach, izolácii DNA a analýze genetických dát, vykonal štatistické analýzy, interpretoval výsledky a napísal štúdiu. B. Mráz P, Garcia-Jacas N, Gex-Fabry E, Susanna A, Barres L, Müller-Schärer H. 2012. Allopolyploid origin of highly invasive Centaurea stoebe sl (Asteraceae). Molecular Phylogenetics and Evolution 62: 612–623. Autorský podiel uchádzača: 70%. PM navrhol a koordionoval štúdiu, nadviazal kontakt so španielskym tímom, podieľal sa na zbere a pestovaní rastlinného materiálu a cytometrických a karyologických analýzach, izolácii DNA a analýze dát, interpretoval výsledky a napísal štúdiu. C. Šingliarová B, Hodálová I & Mráz P. 2011. Biosystematic study of the diploid- polyploid Pilosella alpicola group with variation in breeding system: patterns and processes. Taxon 60: 450–470. Autorský podiel uchádzača: 50% PM navrhol a koordinoval štúdiu, čiastočne sa podieľal na zbere rastlinného materiálu, previedol molekulárne analýzy a významne sa podieľal na interpretácií výsledkov a na písaní textu. D. Šingliarová B, Chrtek J, Plačková
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