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Polygonum Cognatum Meissn. (Madimak) Tohumlarinin

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Polygonum Cognatum Meissn. (Madimak) Tohumlarinin POLYGONUM COGNATUM MEISSN. (MADIMAK) TOHUMLARININ ÇİMLENME B İYOLOJİSİ ve KLONLARI ARASINDAK İ GENETİK ÇEŞİTLİLİĞİN BELİRLENMESİ Mesut ÖZKURT Y.Lisans Tezi Bitki Koruma Anabilim Dalı Doç. Dr. Hüse yin ÖNEN T.C. GAZİOSMANPAŞA ÜNİVERSİTESİ FEN BİLİMLERİ ENSTİTÜSÜ BİTKİ KORUMA ANABİLİM DALI Y.LİSANS TEZİ POLYGONUM COGNATUM MEISSN. (MADIMAK) TOHUMLARININ ÇİMLENME BİYOLOJİSİ ve KLONLARI ARASINDAKİ GENETİK ÇEŞİTLİLİĞİN BELİRLENMESİ Mesut ÖZKURT TOKAT 2008 Her hakkı saklıdır TEZ BEYANI Tez yazım kurallarına uygun olarak hazırlanan bu tezin yazılmasında bilimsel ahlak kurallarına uyulduğunu, başkalarının eserlerinden yararlanılması durumunda bilimsel normlara uygun olarak atıfta bulunulduğunu, tezin içerdiği yenilik ve sonuçların başka bir yerden alınmadığını, kullanılan verilerde herhangi bir tahrifat yapılmadığını, tezin herhangi bir kısmının bu üniversite veya başka bir üniversitedeki başka bir tez çalışması olarak sunulmadığını beyan ederim. Mesut ÖZKURT Doç. Dr. Hüseyin ÖNEN danışmanlığında, Mesut ÖZKURT tarafından hazırlanan bu çalışma 07/04/2008 tarihinde aşağıdaki jüri tarafından oy birliği/oy çokluğu ile Bitki Koruma Anabilim Dalı’nda Yüksek Lisans tezi olarak kabul edilmiştir. Başkan : Prof. Dr. İzzet KADIOĞLU İmza : Üye : Doç. Dr. Hüseyin ÖNEN İmza : Üye : Yrd. Doç. Dr. İskender PARMAKSIZ İmza : Yukarıdaki sonucu onaylarım Prof. Dr. Metin YILDIRIM Enstitü Müdürü ÖZET Y. Lisans Tezi POLYGONUM COGNATUM MEISSN. (MADIMAK) TOHUMLARININ ÇİMLENME BİYOLOJİSİ ve KLONLARI ARASINDAKİ GENETİK ÇEŞİTLİLİĞİN BELİRLENMESİ Mesut ÖZKURT Gaziosmanpaşa Üniversitesi Fen Bilimleri Enstitüsü Bitki Koruma Anabilim Dalı Danışman: Doç. Dr. Hüseyin ÖNEN Türkçede, madımak olarak isimlendirilen Polygonum cognatum Meissn. tarım alanlarında veya tarla sınırları, karayolları kenarları, hava alanları ve endüstriyel bölgeler gibi tarım yapılmayan alanlarda yabancı ot olarak bulunmaktadır. Ancak, Orta Anadolu Bölgesinde (özellikle Tokat, Sivas ve Çorum Amasya illerinde) insanlar tarafından besin olarak yoğun bir şekilde tüketildiğinden, bu yörelerimizde halk kültüründe önemli yer işgal eden bir bitkidir. Bununla birlikte, madımak üzerinde çok az sayıda araştırma yapılmış olup, mevcut araştırmalar da genellikle madımağın ana vatanı, yayılış alanları ile bazı biyolojik özelliklerinin saptanması ve halk tababetinde kullanımı üzerinde yoğunlaşmıştır. Bu çalışmada; i) İç Anadolu’nun farklı ekolojik koşullara sahip bölgelerinden (Tokat, Sivas, Çorum ve Amasya illerinde) toplanan P. cognatum klonları arasındaki genetik varyasyonun RAPD-PCR çalışmaları ile ortaya konması, ii) madımak tohumlarının genel özelliklerinin belirlenmesi ve iii) madımak tohumlarında görülen dormansinin ortadan kaldırılması için uygun yöntemlerin belirlenmesi amaçlanmıştır. RAPD-PCR çalışmasında toplam 45 primer denenmiştir. Daha önce öngörülmeyen problemler nedeniyle bunlardan sadece 4 primerle çalışılmıştır. Tohumların genel özellikleri mikroskop altında belirlenmiş ve tohumların bin dane ağırlığı saptanmıştır. Tohumlarda görülen dormansinin ortadan kaldırılması amacıyla sülfürik asit, gibberellik asit ve potasyum nitrat uygulamaları yapılmış, tohumlar ön üşütme veya ön ısıtmaya tabi tutulmuş ve tohumlara akan suyun altında yıkanma ile mekanik kazıma işlemleri uygulanmıştır. RAPD-PCR çalışmasında kullanılan primerlerdan 80 bant elde edilmiş, bu bantlardaki polimorfizmin % 100 olduğu saptanmıştır. Elde edilen bantlara göre çalışmada kullanılan 68 klon 3 farklı gruba ayrılmıştır. Madımak tohumları üzerinde yapılan çalışmalarda, tohumların achene formunda, üzerinde perinat kalıntıları olan üç i yüzeyli, tohum kabuğu düz, parlak ve koyu kahve-siyah renkli bir meyve olduğu, tohum boyutlarının ortalama 1,64 x 2,94 mm olduğu ve bin dane ağırlığının 3,98 gr olduğu saptanmıştır. Tohum kabuğunun mekanik olarak veya sülfürik asit uygulaması ile aşındırılması tohumların çimlenmesini büyük oranda arttırmıştır. Sülfürik asit ve gibberellik asit uygulamaları beraberce yapıldığında ise en yüksek çimlenme oranı saptanmış ve tohumların % 57’ye varan oranda çimlenebildiği belirlenmiştir 2008, 65 Sayfa Anahtar Kelimeler: Polygonum cognatum, Madımak, Genetik varyasyon, RAPD, PCR, Dormansi, Tohum çimlenme biyolojisi ii ABSTRACT Ms Thesis DETERMINATION OF GERMINATION BIOLOGY OF MADIMAK (POLYGONUM COGNATUM MEISSN.) SEEDS AND GENETIC VARIATION AMONG CLONES Mesut OZKURT Gaziosmanpaşa University Graduate School of Natural and Applied Sciences Department of Plant Protection Supervisor: Assoc. Prof. Dr. Huseyin ONEN Polygonum cognatum Meissn, known as ‘madimak’ in Turkish, is a weed found in both agricultural and non-agricultural areas such as field edges and roadsides, airfield and industrial areas. Since the weed is consumed as vegetable, especially in Central Anatolian Region, it is also an important folkloric plant. Literature review shows that number of researches on madimak is very limited. The researches generally focused on the origin and distribution of madimak, and determined some of biological characteristic and the process of consuming as a vegetable and folk medicine plant. The aims of this research are to; i) investigate genetic variation among P. cognatum clones adapted to different ecological conditions of Central Anatolian (Tokat, Sivas, Corum and Amasya provinces) Region using RAPD-PCR analyses, ii) determine seed properties of P. cognatum, and iii) determine suitable processes to overcome seed dormancy. Forty-five primers were tested for the RAPD-PCR analyses. Due to unexpected problems occurred during the experiment, only four primers were selected to identify the genetic distances among madimak clones. Sulfuric acid, hot water and mechanical seed scarifications, potassium nitrate, gibberellik acid, chilling applications, and washing under running water treatments were used to promote seed germination. Totally 80 bands were obtained from the RAPD-PCR studies. The polymorphic rate of the bands was 100 %. Depending on pattern of bands, total 68 clones collected from different habitats were divided into three groups. The fruit of madiamak (a triangular achene surrounded by remains of perianth, dark brown or black in color, smooth and shiny) was measured as 1.64 mm x 2.94 mm in iii size, in average. The average 1000 seeds weight was determined as 3.98 g. Scarification in sulfuric acid and mechanically scarification greatly increased germination. Together scarification in sulfuric acid and gibberellik acid application resulted in the highest seed germination (up to 57 %). 2008, 65 Pages Keywords: Polygonum cognatum Meissn., Madimak, Genetic variation, RAPD, PCR, Dormansi, Seed germination biology. iv ÖNSÖZ Çalışmamın her aşamasında bilgi, öneri, yardım ve desteğini esirgemeyen ayrıca engin fikirleriyle akademik anlamda yetişme ve gelişmeme katkıda bulunan danışman hocam Sayın Doç. Dr. Hüseyin ÖNEN’e, ayrıca çalışmalarımın her aşamasında yardımlarını esirgemeyen ve bana maddi manevi destek olan Sayın Yrd. Doç. Dr. İskender PARMAKSIZ’a, bilimsel yaklaşımları ve engin bilgileri ile gerek çalışmalarımda gerekse yetişmemde yardımı olan bölüm başkanımız Sayın Prof. Dr. İzzet KADIOĞLU ve diğer bölüm hocalarıma, çalışmalarımın başından sonuna kadar benden yardımlarını esirgemeyen arkadaşlarım Zir. Yük. Müh. Melih YILAR, Zir. Yük. M. Yaşar ÖZDEMİR, Zir. Yük. Fatih ALAY, Yüksek Lisan Öğrencisi Murat AYDEMİR, Yüksek Lisan Öğrencisi İsmail BENLİ’ye teşekkürü bir borç bilirim. Ayrıca tüm hayatım boyunca attığım her adımda benden hiçbir fedakârlığı esirgemeyen aileme ve çalışmalarımın her aşamasında manen desteğini esirgemeyen, benim stres ve yorgunluğuma katlanan arkadaşlarıma tüm yakınlarıma bütün samimi duygularımla teşekkür ederim. Bu tez; TÜBİTAK TOVAG (Tarım Ormancılık Veterinerlik Araştırma Gurubu) tarafından desteklen “Orta Anadolu Orijinli Madımak (Polygonum cognatum Meissn.) Genotiplerinin Moleküler ve Agronomik Özellikleri ile Allelopatik Potansiyellerinin Belirlenmesi” adlı ve 105 O 502 nolu proje kapsamında yürütülen çalışmaların bir kısmını içermektedir. Mesut ÖZKURT Mayıs 2008 v İÇİNDEKİLER Sayfa ÖZET ……........................................................................................................................I ABSTRACT......................................................................................................................III ÖNSÖZ …………………...…........................................................................................V İÇİNDEKİLER………………………………………………………………………….VI RESİMLER DİZİNİ........................................................................................................IX ÇİZELGELER DİZİNİ…………………………………………………………………X SİMGE ve KISALTMALAR DİZİNİ.............................................................................XI 1.GİRİŞ…………………………………………………………………………………..1 2. LİTERATÜR ÖZETLERİ……………………………………………………………..3 2.1. Polygonaceae Familyasının Genel Özellikleri……………………………………….3 2.2 Polygonaceae Cinsinin Genel Özelikleri……………….…………………………….3 2.3 Polygonaceae Cinsinin Sistematiği……………………..……………………………4 2.4. Polygonum cognatum (Meissn.)’ in Genel Özellikleri..……………………………..4 2.4.1. Çevre İstekleri……………………………………………………………………...5 2.4.2. Bitkinin İçeriği……………………………………………………………………..5 2.4.3. Kullanılma Şekiller………………………………………………………………...5 2.5. Çimlenme Çalışmaları……………………………………………………………….5 2.6. DNA Markörleri……………………………………………………………………..8 3. MATERYAL ve YÖNTEM………………………………………………………….17 3.1. Materyal…………………………………………………………………………….17 3.2. Yöntem……………………………………………………………………………..22 3.2.1. Madımak Tohumlarının Bazı Morfolojik Özelliklerinin Belirlenmesi…………..22 3.2.1.1. Tohum
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