Dergiler / EurAsian Journal of BioSciences (elektronik) / 2010 / Sayı: 4
Molecular variability among the isolates of Sclerotium rolfsii causing stem rot of groundnut by RAPD, ITS-PCR and RFLP
- Sayı
- 2010 · Sayı: 4
- Sayfa
- 80–87
- DOI
- —
Özet
Sclerotium rolfsii'nin virulent izolatlari arasindaki genetik çesitlilik RAPD, ITS-PCR ve RFLP gibi moleküler teknikler kullanilarak çalisildi. RAPD bantlari, iki küme olusturmak suretiyle izolatlar arasindaki genetik çesitliligi gösterdi. Bes RAPD primeri kullanilarak, yaklasik 100 bp ile 2500 bp arasinda degisiklik gösteren toplam 221 adet tekrarlanabilir ve ölçülebilir bant elde edildi. Spesifik ITS1 ve ITS4 universal primerleri ile rDNA'nin ITS bölgesinin çogatilmasi, bütün izolatlarda yaklasik 650 ila 700 bp üretti ve bu durum bütün izolatlarin S. rolfsii'den elde edildigini teyit etti. ITS-RFLP çalismalari, kullanilan restrüksiyon endonükleazlarla izolatlar arasindaki restrüksiyon bantlarinda her hangi bir polimorfizm olmadigini gösterdi.
Abstract
Genetic variability among the virulent isolates of Sclerotium rolfsii was studied using molecular techniques like RAPD, ITS-PCR and RFLP. The RAPD banding pattern reflected the genetic diversity among the isolates by formation of two clusters. A total of 221 reproducible and scorable polymorphic bands ranging approximately as low as 100 bp to as high as 2500 bp were generated with five RAPD primers. ITS region of rDNA amplification with specific ITS1 and ITS4 universal primers produced approximately 650 to 700 bp in all the isolates confirmed that all the isolates obtained are Sclerotium rolfsii. Studies by ITS-RFLP indicated that there was no polymorphism in restriction banding pattern among the isolates with the restriction endonucleases used.