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Download (1383Kb) UNIVERZA V LJUBLJANI PEDAGOŠKA FAKULTETA BIOTEHNIŠKA FAKULTETA FAKULTETA ZA KEMIJO IN KEMIJSKO TEHNOLOGIJO NAROVOSLOVNOTEHNIŠKA FAKULTETA Študijski program: Kemija in Biologija UGOTAVLJANJE SORODSTVENIH ODNOSOV V PODRODU METULJEV AGRODIAETUS NA BALKANSKEM POLOTOKU DIPLOMSKO DELO INFERRING THE RELATIONSHIP BETWEEN BUTTERFLY SPECIES IN THE SUBGENUS AGRODIAETUS ON BALKAN PENINSULA GRADUATION THESIS Mentor: dr. Rudi Verovnik Kandidat: Tom Koritnik LJUBLJANA, oktober, 2014 Koritnik T. Ugotavljanje sorodstvenih odnosov v podrodu metuljev Agrodiaetus I na Balkanskem polotoku. Diplomsko delo. Ljubljana, Univerza v Ljubljani, Pedagoška fakultera, Biologija in Kemija, 2014. Diplomsko delo je zaključek univerzitetnega študijskega programa Kemija in Biologija. Opravljeno je bilo na Katedri za zoologijo Oddelka za biologijo Biotehniške fakultete Univerze v Ljubljani. Komisija za dodiplomski študij Oddelka za biologijo je za mentorja diplomskega dela imenovala doc. dr. Rudija Verovnika. Komisija za oceno in zagovor: Predsednik: doc. dr. Primoţ ZIDAR Univerza v Ljubljani, Biotehniška fakulteta Član: doc. dr. Cene FIŠER Univerza v Ljubljani, Biotehniška fakulteta Mentor: doc. dr. Rudi VEROVNIK Univerza v Ljubljani, Biotehniška fakulteta Datum zagovora: Podpisani se strinjam z objavo svojega diplomskega dela v polnem tekstu na spletni strani Digitalne knjiţnice Biotehniške fakultete. Izjavljam, da je diplomsko delo, ki sem ga oddal v elektronski obliki, identično tiskani verziji. Delo je rezultat lastnega raziskovalnega dela. Tom Koritnik Koritnik T. Ugotavljanje sorodstvenih odnosov v podrodu metuljev Agrodiaetus II na Balkanskem polotoku. Diplomsko delo. Ljubljana, Univerza v Ljubljani, Pedagoška fakultera, Biologija in Kemija, 2014. KLJUČNA DOKUMENTACIJSKA INFORMACIJA ŠD Dn DK KG metulji /Lepidoptera /modrini /Polyommatus /Agrodiaetus /Polyommatus admetus /Polyommatus ripartii /Polyommatus eleniae /Polyommatus aroaniensis /Polyommatus orphicus /kriptične vrste /PCR /filogenetska analiza /morfologija kril /fenotipski znaki AV KORITNIK, Tom SA VEROVNIK, Rudi (mentor) KZ SI-1261Dobrunje, Pot v Podgorje 3a ZA Univerza v Ljubljani, Pedagoška fakulteta, Biotehniška fakulteta LI 2014 IN UGOTAVLJANJE SORODSTVENIH ODNOSOV V RODU METULJEV AGRODIAETUS NA BALKANSKEM POLOTOKU TD Diplomsko delo (univerzitetni študij) OP VII, 45 str., 3 preg., 6 sl., 49 virov IJ sl JI sl/en AI V diplomskem delu smo raziskovali sorodstvene odnose in vrstno pripadnost v podrodu modrinov Polyommatus (subgenus Agrodiaetus) na Balkanskem polotoku. Metulje smo predhodno določili na podlagi fenotipskih znakov. Vse osebke smo nato poskusili tudi natančneje uvrstiti glede na širši nabor morfoloških znakov, ki se uporabljajo v taksonomiji tega podroda. Vrstno pripadnost smo ugotavljali na podlagi primerjave zaporedij mitohondrijskega gena COI, ki se uporablja kot genetska črtna koda. Filogenetske odnose smo ugotavljali z Bayesovo analizo in jih primerjali z morfološkimi določitvami osebkov in njihovo razširjenostjo. Glede na fenotipske znake smo lahko z gotovostjo uvrstili le osebke vrste Polyommatus admetus, pri ostalih vrstah pa je variabilnost fenotipskih znakov prevelika za nedvoumno določitev. Filogenetski rezultati so potrdili prisotnost vsaj petih vrst (P. admetus, P. ripartii, P. aroaniensis, P. orphicus in P. eleniae) med našimi vzorci. Glede na geografsko razširjenost osebkov posameznih vrst nismo potrdili alopatrije oţje sorodnih vrst. Simpatrija med bolj sorodnimi vrstami kaţe na reproduktivne bariere tudi med morfološko zelo podobnimi vrstami. Ugotovili smo tudi, da je območje razširjenosti vrste P. orphicus večje, kot je navedeno v literaturi. Koritnik T. Ugotavljanje sorodstvenih odnosov v podrodu metuljev Agrodiaetus III na Balkanskem polotoku. Diplomsko delo. Ljubljana, Univerza v Ljubljani, Pedagoška fakultera, Biologija in Kemija, 2014. KEY WORDS DOCUMENTATION (KWD) DN dn DC CX butterflies /Lepidoptera /blues /Polyommatus /Agrodiaetus /Polyommatus admetus / Polyommatus ripartii /Polyommatus eleniae /Polyommatus aroaniensis /Polyommatus orphicus /cryptic species /PCR /philogenetic analisys /wing morphology /fenotype characteristics AU KORITNIK, Tom AA VEROVNIK, Rudi (supervisor) PP SI-1261 Dobrunje, Pot v Podgorje 3a PB University of Ljubljana, Faculty of Education, Biotechnical faculty PY 2014 TI Inferring the relationship between butterfly species in the subgenus Agrodiaetus on Balkan Peninsula DT Graduation Thesis (University studies) NO VII, 45 p., 3 tab., 6 fig., 49 ref. LA sl AL sl/en AI We studied the phylogenetic relationships in the subgenus of the Polyommatus (subgenus Agrodiaetus) butterflies in the Balkan Peninsula. Preliminarily identifications of all specimens were based on their wing morphology. In turn, all individuals were examined for a larger number of morphological characters, that proved useful in taxonomy of the subgenus. Phylogenetic relationships were reconstructed using the mitochondrial DNA gene COI, used as a DNA barcodes. We reconstructed phylogeny with the Bayesian analysis and then compared these results with the morphological characteristics and distribution ranges of studied species. Only Polyommatus admetus can be unambigously identified using morphological traits; the identiy of other herein studied species cannot be inferred from morphology alone due to high intraspecific variation. The phylogentic confirmed the presence of at least five species (P. admetus, P. ripartii, P. aroaniensis, P. orphicus in P. eleniae) in our samples. According to the geographical distribution of the samples, we could not confirm alopatry of closely related species. Simpatry between more closely related species suggests that even morphologically similar species are reproductively isolated. We also found that the distribution range of the P. orphicus species is larger than previously thought. Koritnik T. Ugotavljanje sorodstvenih odnosov v podrodu metuljev Agrodiaetus IV na Balkanskem polotoku. Diplomsko delo. Ljubljana, Univerza v Ljubljani, Pedagoška fakultera, Biologija in Kemija, 2014. Kazalo vsebine KLJUČNA DOKUMENTACIJSKA INFORMACIJA ............................................................ II KEY WORDS DOCUMENTATION (KWD) ......................................................................... III Kazalo vsebine ......................................................................................................................... IV Kazalo preglednic: .................................................................................................................... VI Kazalo slik: ............................................................................................................................... VI Okrajšave ................................................................................................................................. VII 1 UVOD ................................................................................................................................ 1 2 PREGLED OBJAV ............................................................................................................ 2 2.1 Ţuţelke ........................................................................................................................ 2 2.2 Metulji ......................................................................................................................... 2 2.3 Druţina modrinov (Lycaenidae) .................................................................................. 4 2.4 Rod Polyommatus ........................................................................................................ 5 2.5 Podrod Agrodiaetus ..................................................................................................... 5 2.5.1 Polyommatus admetus (Esper, 1783) ................................................................... 6 2.5.2 Polyommatus ripartii (Freyer, 1830) ................................................................... 6 2.5.3 Polyommatus aroaniensis (Brown, 1976) ............................................................ 7 2.5.4 Polyommatus eleniae (Coutsis in De Prins, 2005) ............................................... 7 2.5.5 Polyommatus orphicus (Kolev, 2005) .................................................................. 7 2.6 Kriptične vrste ............................................................................................................. 8 2.7 Črtne kode DNK .......................................................................................................... 8 2.8 Reakcija PCR ............................................................................................................... 9 3 MATERIALI IN METODE ............................................................................................. 10 3.1 Vzorci ........................................................................................................................ 10 Koritnik T. Ugotavljanje sorodstvenih odnosov v podrodu metuljev Agrodiaetus V na Balkanskem polotoku. Diplomsko delo. Ljubljana, Univerza v Ljubljani, Pedagoška fakultera, Biologija in Kemija, 2014. SEZNAM VZORCEV IN LOKALITET ......................................................................... 11 3.2 Morfologija kril ......................................................................................................... 13 3.3 MOLEKULARNOBIOLOŠKA ANALIZA ............................................................. 15 3.3.1 Kemikalije, potrošni material in naprave ..........................................................
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