Biologia plantarum 54:435-442, 2010 | DOI: 10.1007/s10535-010-0079-y

PCR-based molecular markers for assessment of somaclonal variation in Pinus pinea clones micropropagated in vitro

C. Cuesta1, R. J. Ordás1, A. Rodríguez1, B. Fernández1,*
1 Área de Fisiología Vegetal, Dpto. BOS, Instituto de Biotecnología de Asturias, Universidad de Oviedo, Oviedo, Spain

Four different markers [random amplified polymorphic DNA (RAPD), inter simple sequence repeat (ISSR), amplified fragment length polymorphism (AFLP), and selective amplified microsatellite polymorphism length (SAMPL)] were applied for evaluating somaclonal variation of micropropagated genotypes of stone pine (Pinus pinea L.). The total number of primers tested was 130, with 223 combinations assayed. A high number of them amplified successfully (178), representing 79.82 % of the total, and the average number of amplified fragments ranged from 2.47 (ISSR) to 65.76 (SAMPL). Based on internal controls, no problem of reproducibility was detected. Almost no somaclonal variation was detected within the clones. Of the tested markers, ISSR, AFLP, and SAMPL showed monomorphic amplification profiles, with only RAPD markers showing some interclonal variation.

Keywords: AFLP; caulogenesis; clonal stability; conifers; ISSR; RAPD; SAMPL; stone pine
Subjects: amplified fragment length polymorphism (AFLP); ex vitro transfer; genetic diversity; inter-simple sequence repeat (ISSR); in vitro culture, direct organogenesis; pine; Pinus pinea; selective amplified microsatellite polymorphism length (SAMPL)

Received: July 20, 2008; Accepted: May 27, 2009; Published: September 1, 2010Show citation

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Cuesta, C., Ordás, R.J., Rodríguez, A., & Fernández, B. (2010). PCR-based molecular markers for assessment of somaclonal variation in Pinus pinea clones micropropagated in vitro. Biologia plantarum, 54(3), 435-442. doi: 10.1007/s10535-010-0079-y.
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