Cevizde Yeni SSR Markörlerin Geliştirilmesi

Year 2016, Volume: 45 Issue: (Özel Sayı 1) 7. Ulusal Bahçe Bitkileri Kongresi, 207 - 211, 31.03.2016

Hayat Topçu , Adi Surya Ikhsan Mehmet Sütyemez , Nergiz Çoban Murat Güney , Salih Kafkas

Abstract

Bu çalışmada ceviz için yeni SSR markörlerin geliştirilmesi amaçlanmıştır. Bunun için Maraş-18 ceviz çeşidi DNA’sı kullanılarak oluşturulan genomik kütüphane dört farklı motif ile (CA, GA, AAC ve AAG) zenginleştirilmiştir. Sekansı yapılan 620 klondan 276 adet primer çifti dizayn edilmiştir. Öncelikle gradient PCR ile her bir primer çiftinin DNA’ya bağlanma sıcaklıkları belirlenmiştir. Bant veren 247 SSR primer çifti 15 ceviz çeşidinde analiz edilerek polimorfizm durumları belirlenmiştir. Toplam 185 primer çifti polimorfizm göstermiş olup bunların 80 tanesi (%44.4) GA kütüphanesinden elde edilmiştir. En az polimorfik primer çifti (14 adet) ise AAC kütüphanesinden elde edilmiştir. CA ve AAG kütüphanelerinden ise benzer oranda polimorfizm elde edilmiştir. Sonuç olarak, cevizde bundan sonra yapılacak genetik kaynakların karakterizasyonu, genetik haritalama, popülasyon genetiği gibi çalışmalarda kullanmak üzere çok sayıda yeni SSR markörleri geliştirilmiştir.

Keywords

Ceviz , Juglans regia , SSR , mikrosatellit

Development of Simple Sequence Repeat (SSR) Markers from Walnut (Juglans regia L.)

Abstract

In this study, it was aimed to develop SSR markers for walnut. Genomic libraries enriched with CA, GA, AAC and AAG repeats were constructed using genomic DNA of Juglans regia cv. Maraş-18 to develop SSR markers for walnut. A total of 624 clones were sequenced and 276 primer pairs were designed. Amplification of SSR primer pairs was tested by gradient PCR to determine their optimum annealing temperatures, 247 generated PCR products, and 185 polymorphic loci were obtained by characterizing 15 walnut cultivars. A total of 185 primer pairs showed polymorphism and 80 of them obtained from GA-enriched library. The least polymorphism (14 primer) obtained from AAC-enriched library. CA and AAG enriched libraries produced similar polymorphism rates. As a result, the new SSR primer pairs developed in this study can be used in different genetic studies in walnut such as characterization of genetic resources, construction of genetic linkage maps, and population genetics.

Keywords

Walnut , Juglans regia , SSR , microsatellite

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