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Asteraceae) Kompleksine Ait Tür Populasyonlarinda Genetik Çeşitliliğin Belirlenmesi

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Asteraceae) Kompleksine Ait Tür Populasyonlarinda Genetik Çeşitliliğin Belirlenmesi T.C. SELÇUK ÜNİVERSİTESİ FEN BİLİMLERİ ENSTİTÜSÜ PSEPHELLUS AUCHERIANUS (DC.) BOISS. (ASTERACEAE) KOMPLEKSİNE AİT TÜR POPULASYONLARINDA GENETİK ÇEŞİTLİLİĞİN BELİRLENMESİ MERYEM BOZKURT DOKTORA TEZİ Biyoloji Anabilim Dalını Ağustos-2015 KONYA Her Hakkı Saklıdır TEZ BİLDİRİMİ Bu tezdeki bütün bilgilerin etik davranış ve akademik kurallar çerçevesinde elde edildiğini ve tez yazım kurallarına uygun olarak hazırlanan bu çalışmada bana ait olmayan her türlü ifade ve bilginin kaynağına eksiksiz atıf yapıldığını bildiririm. DECLARATION PAGE I hereby declare that all information in this document has been obtained and presented in accordance with academic rules and ethical conduct. I also declare that, as required by these rules and conduct, I have fully cited and referenced all material and results that are not original to this work. Meryem BOZKURT 07.08.2015 ÖZET DOKTORA TEZİ PSEPHELLUS AUCHERIANUS (DC.) BOISS. (ASTERACEAE) KOMPLEKSİNE AİT TÜR POPULASYONLARINDA GENETİK ÇEŞİTLİLİĞİN BELİRLENMESİ Meryem BOZKURT SELÇUK ÜNİVERSİTESİ FEN BİLİMLERİ ENSTİTÜSÜ BİYOLOJİ BÖLÜMÜ ANABİLİM DALI Danışman: Doç. Dr. Tuna UYSAL 2015, 157 Sayfa Jüri Danışman: Doç. Dr. Tuna UYSAL Prof. Dr. Kuddisi ERTUĞRUL Prof. Dr. Leyla AÇIK Prof. Dr. Muhittin DİNÇ Doç. Dr. Emine ARSLAN Bu tez çalışmasıyla Psephellus aucherianus ve yakın akraba olduğu düşünülen Ps. sintenisii ve Ps. yusufeliensis taksonları morfolojik, mikromorfolojik, karyolojik ve moleküler metotlar ile incelenmiş taksonların karakterizasyonu yapılarak, akrabalık ilişkileri ve populasyonların genetik yapıları detaylı bir biçimde ele alınmıştır. Taksonomik olarak yakın akraba takson veya grupların her ne kadar sınırlarını belirlemek kolay olmasada elde edilen veriler doğrultusunda Ps. sintenisii’nin Ps. aucherianus orjinli olduğu ve genetik sürüklenmenin etkisiyle serpantine adapte olarak hem genetik hem de taksonomik açıdan önemli derecede farklılaştığı belirlenmiştir. Uygulanan moleküler markırlar Ps. aucherianus ve Ps. yusufeliensis taksonlarının aynı soydan türemiş yakın akraba taksonlar olduğuna işaret etmiş, karyolojik ve moleküler verilerin yanısıra küçük morfolojik farklılıklara dayalı olarak taksonlar karakterize edilmiştir. SEM analizlerinden gelen aken mikromorfolojisi verilerine göre en farklılaşmış takson olarak Ps. sintenisii gözükmektedir. Karyolojik veriler yakın akraba taksonların oldukça benzer karyomorfolojik özelliklere sahip olduğunu bildirmektedir. Karyotip formülleri türlere özgün olup taksonomik ayrım yapmaya imkân vermektedir. Buna göre türler diploid kromozom sayısına sahip olup (2n=30), cinse ait özgün temel kromozom sayısını (x=15) içermektedir. Moleküler veriler temel olarak ele alınan üç taksonun ortak bir orjinden kaynaklandığını ve belli gen bölgeleri açısından farklılaşmış olduklarına işaret etmekte, belirli bariyerlerin etkisi altında hızla farklılaşarak evrilmeye devam ettiklerini göstermektedir. Bununla birlikte AFLP ve matK’dan gelen bilgiler populasyon bazında genetik varyasyon seviyelerinin düşük olmasının taksonların geleceğine yönelik bir riskin varlığına dikkat çekmektedir. Buna bağlı olarak, Ps. aucherianus’a ait üç, Ps. yusufeliensis’e iki ve tek tip lokaliteden bilinen Ps. sintenisii’ye ait populasyonlar üzerinde yapılan moleküler analizler her bir populasyon arası genetik farklılaşmanın yüksek olduğunu açığa çıkarmıştır. Sonuç olarak nadir endemik türler için genel olarak öngürüldüğü gibi ele alınan taksonların genetik çeşitlilik seviyelerinin düşük olması sebebiyle, insan etkisiyle olmasa bile doğal etkiler nedeniyle nesli tükenme riski ile karşı karşıya oldukları belirlenmiştir. Oldukça izole yayılış gösteren Ps. aucherianus, Ps. sintenisii ve Ps. yusufeliensis taksonlarının nispeten düşük tohum ve polen yayma kapasiteleri, kendileşmeye uygun üreme davranışları ve genetik süreklenmenin neden olduğu gen havuzundaki fakirleşmeler (genetik yapı ve populasyonların ileri seviyede farklılaşması) sonucunda lokal bir nesli tükenme tehditi ile karşı karşıya oldukları ortaya çıkarılmıştır. Populasyon gözlemlerimiz ve moleküler analizlerimizden ortaya çıkan değerlendirmelere göre Uluslararası IUCN kriteleri dikkate alındığında Ps. sintenisii ve Ps. yusufeliensis CR (Critically Endangered/ Kritik Tehlikede), Ps. aucherianus ise EN (Endangered/ Tehlikede) olarak değerlendirilmiştir. iv Anahtar Kelimeler: cpDNA haplotip, endemik, genetik çeşitlilik, karyomorfoloji, koruma, meyve yüzeyi, moleküler markırlar, Türkiye. v ABSTRACT Ph. D THESIS DETERMINATION OF THE GENETIC VARIATION IN POPULATIONS BELONGING TO PSEPHELLUS AUCHERIANUS (DC.) BOISS. (ASTERACEAE) COMPLEX SPECIES Meryem BOZKURT THE GRADUATE SCHOOL OF NATURAL AND APPLIED SCIENCE OF SELÇUK UNIVERSITY THE DEGREE OF DOCTOR OF PHILOSOPHY IN BIOLOGY Advisor: Assoc. Prof. Dr. Tuna UYSAL 2015, 157 Pages Jury Advisor: Assoc. Prof. Dr. Tuna UYSAL Prof. Dr. Kuddisi ERTUĞRUL Prof. Dr. Leyla AÇIK Prof. Dr. Muhittin DİNÇ Assoc. Prof. Dr. Emine ARSLAN In this thesis, the relationship between Psephellus sintenisii and Ps. yusufeliensis, which are assumed to be closely related to Ps. aucherianus taxa, and genetic structures of their populations have been investigated in detail by characterizing taxa morphologically, micromorphologically, karyologically and molecular methods. Although it is not easy to detect the limits of closely related taxa or groups taxonomically, it is found out that Ps. sintenisii is originated from Ps. aucherianus, and with the effect of genetic drift it is differed from Ps. aucherianus significantly by adapting serpentine both genetically and taxonomically, using obtained data. Applied molecular markers have shown that Ps. aucherianus, and Ps. yusufeliensis taxa are closely related and have originated from the same ancestry. In addition to karyologic and molecular data, taxa have been characterized based on the minor morphological differences. According to the achene micromorphology data obtained from SEM analyses, it is found that Ps. sintenisii is the most differentiated taxon. Karyologic data show that closely related taxa have highly similar karyomorphologic properties. Karyotype formulas are specific for the species, and they give oppurtunity to separate species taxonomically. According to this, the species have diploid chromosomes (2n=30), and have the specific basic chromosome number (x=15) for the genus. Molecular data show that three taxa come from the same origin and differentiate in certain gene regions, and they continue to evolve rapidly under the effect of certain barriers. Besides, data obtained using AFLP and matK point out that there is a risk about the future of taxa because of low genetic variation level on population basis. According to the molecular analyses made using three populations of Ps. aucherianus, two populations of Ps. yusufeliensis, and one population of Ps. sintenisii reveal that there is a high genetic differentiation between each population. In conclusion, as generally predicted for rare endemic species, low genetical variation levels of the taxa under investigation, Ps. aucherianus, Ps. sintenisii, and Ps. yusufeliensis taxa are under the risk of extinction, not because of human effect, but because of natural reasons. As a result of relatively low seed and polen spreading capacity of Ps. aucherianus, Ps. sintenisii, and Ps. yusufeliensis taxa, production behavior appropriate for inbreeding, and impoverishment caused by genetic drift in the gene pool (differentiation of genetic structures and populations in advanced level), it is understood that these taxa, which spread isolatedly are under the local risk of extinction. According to the evaluation resulted from population observations and molecular analyses, when international IUCN criteria are taken vi into consideration, Ps. sintenisii, Ps. yusufeliensis should be categorized as CR (Critically Endangered). Ps. aucherianus are considered as EN (Endangered), respectively. Keywords: Conservation, cpDNA haplotype, endemic, fruit surface, genetic diversity, karyomorphology, molecular markers, Turkey. vii ÖNSÖZ Tez çalışmamın her aşamasında yol gösteren, engin bilgi ve tecrübesinden, yönlendirici katkılarından faydalandığım ve her türlü zorlukta verdikleri büyük destek için danışman hocam sayın Doç. Dr. Tuna UYSAL’a en içten saygı ve şükranlarımı sunarım. Çalışma materyallerinin elde edilmesinde ve çalışma sürecim boyunda hiçbir zaman yardımını esirgemeyen ve her zaman destek olan, bilgi birikimini benimle paylaşan ve tez izleme komitemde bulunan sayın hocam Prof. Dr. Kuddisi ERTUĞRUL’a içtenlikle teşekkürlerimi sunarım. Tez izleme komitemde bulunan ve çalışmamın yönlendirilmesine katkı sağlayan sayın hocam Prof. Dr. Leyla AÇIK’a teşekkürlerimi sunarım. Taramalı elektron mikroskop analizleri sonucu elde edilen verilerin analizinde yardımcı olan sayın hocam Doç. Dr. Feyza Candan’a teşekkür ederim. Laboratuvar çalışmalarım sırasında yardımcı olan arkadaşım Arş. Gör. Ela Nur ŞİMŞEK SEZER’e ve Betül Sena TEKKANAT’a teşekkür ederim. Eğitim hayatım boyunca benden maddi manevi desteğini ve inancını esirgemeyen, çalışma süresince büyük anlayış gösteren ve her zaman her durumda bana verdikleri güven ve sonsuz destek için babam Hasan Hüseyin BOZKURT’a, annem Fatma BOZKURT’a, ağabeyim İsmail BOZKURT’a, ablam SİBEL ERKUL’a ve yeğenlerime en derin teşekkürlerimi sunarım. Meryem BOZKURT KONYA-2015 viii İÇİNDEKİLER ÖZET .............................................................................................................................. iv ABSTRACT ...................................................................................................................
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