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Optimization of DNA isolation and PCR protocol for analysis and evaluation of genetic diversity of the medicinal plant, Anemopsis californica using RAPD

Lizette Del-Toro-Sánchez, C. and Villaseñor-Alvarado, S. and Zurita-Martínez, Florentina and Castellanos-Hernández, O. and Rodríguez-Sahagún, Araceli and Isabel Torres-Morán, M. and Rojas-Bravo, D. and Gutiérrez-Lomelí, M. (2013) Optimization of DNA isolation and PCR protocol for analysis and evaluation of genetic diversity of the medicinal plant, Anemopsis californica using RAPD. Acta Biologica Hungarica, 64 (2). pp. 184-195. ISSN 0236-5383

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Abstract

Anemopsis californica is a perennial herbaceous plant that has been utilized as a medicinal plant for the treatment of various diseases. The present work was carried out with the objective of optimizing a method of extraction of the genomic DNA of A. californica and a PCR protocol and later to evaluate the existing genetic diversity among the genotypes deriving from different origins. For DNA extraction, we tested four procedures: with the CTA B-2 protocol, we obtained the highest yield (61.5±2.2 μg DNA/g of leaf tissues) and the best quality (A<sub>260/280</sub> 1.83±0.022). To estimate genetic variability, we utilized the randomly amplified polymorphism DNA (RAPD) technique, employing 20 oligonucleotides, of which only 18 generated reproducible banding patterns, producing 123 polymorphic bands generated, thus obtaining a polymorphism rate of 93.93% among the genotypes analyzed. The Jaccard similarity coefficient generated a variation ranging from 0.325–0.921, indicating a high level of genetic variation among the studied genotypes. An Unweighted pair-group method with arithmetic mean (UPGMA) group analysis indicated six distinct groups. The present optimized method for DNA isolation and RAPD protocol may serve as an efficient tool for further molecular studies.

Item Type: Article
Subjects: Q Science / természettudomány > QH Natural history / természetrajz
Depositing User: Ágnes Sallai
Date Deposited: 20 Apr 2017 06:52
Last Modified: 20 Apr 2017 06:52
URI: http://real.mtak.hu/id/eprint/51506

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