IDENTIFICATION OF GENETIC DIVERSITY AMONG Arnica montana L. GENOTYPES USING RAPD MARKERS
Sylwia Okoń
University of Life Science in LublinEdyta Paczos-Grzęda
University of Life Science in LublinMagdalena Łoboda
University of Life Science in LublinDanuta Sugier
University of Life Science in LublinAbstrakt
Arnica montana L. is one of the most important herbal plants used in medicine, pharmaceutical and cosmetic industry. The number of studies performed with molecular markers on arnica genotypes is very limited. Because of this fact the aims of presented examination were optimization of protocols DNA isolation from fresh leaves of A. montana and identification of genetic diversity among this plant genotypes. In presented study to obtain pure DNA Plant & Fungi DNA Purification Kit (EURx) were used. To clean obtained DNA long and slow electrophoresis and isolation DNA from gels were used. A. montana genotypes were analyzed using 40 RAPD primers (Operon Technologies), out of which 12 produced high number of polymorphic and repeatable fragments. In total, selected primers produced 120 fragments, among them 111 (92.5%) were polymorphic. The genetic similarity matrices were produced based on RAPD using the Dice’s coefficient.
RAPD based genetic similarity was estimated between 0.535 and 0.945. The highest genetic similarity was estimated among GA17 and GA18 genotypes, which are closely located on the obtained dendrogramme.
Słowa kluczowe:
DNA polymorphism, medicinal plant, molecular analysisBibliografia
Belaj A., Trujillo I., de la Rosa R., Rallo L., Giménez M.J., 2001. Polymorphism and discrimination capacity of randomly amplified polymorphic markers in olive germplasm bank. J. Amer. Soc. Hort. Sci., 126(1), 64–71.
Bilia A.R., Bergonzi M.C., Mazzi G., Vincieri F.F., 2006. Development and stability of semisolid preparations based on a supercritical CO2 Arnica extract. J. Pharm. Biomed. Anal., 41, 449–454.
Chen L., Gao Q., Chen D., Xu C., 2005. The use of RAPD markers for detecting genetic diversity, relationship and molecular identification of Chinese elite tea genetic resources (Camellia sinensis (L.) O. Kuntze) preserved in a tea germplasm repository. Biodivers. Conserv., 14(6), 1433–1444.
Doyle J.J., Doyle J.L., 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull., 19, 11–15.
Gawlik-Dziki U., Świeca M., Sugier D., Cichocka J., 2011. Comparison of in vitro lipoxygenase, xanthine oxidase inhibitory and antioxidant activity of Arnica montana and Arnica chamissonis tinctures. Acta Sci. Pol., Hortorum Cultus, 10(3), 15–27.
Hadian J., Tabatabaei S.M.F., Naghavi M.R., Jamzad Z., Ramak-Masoumi T. 2007. Genetic diversity of Iranian accessions of Satureja hortensis L. based on horticultural traits and RAPD markers. Sci. Hortic, 115(2), 196–202.
Hammer Ø., Harper D.A.T., Ryan P.D., 2001. Past: paleontological statistics software package for education and data analysis. Palaeont. Electron., 4(1) 4, 1–9.
Hosokawa K., Minami M., Kawahara K., Nakamura I., Shibata T., 2000. Discrimination among tree spece sof medicinal Scutellaria plants Rusing RAPD markers. Planta Med., 66, 270–272.
Kapteyn J., Goldsbrough P.B., Simon J.E., 2002. Genetic relationships and diversity of commercially relevant Echinacea species. Theor. Appl. Genet., 105(2–3), 369–376.
Khanuja S.P.S., Shasany A.K., Srivastava A., Kumar S., 2000. Assessment of genetic relationships in Mentha species. Euphytica, 111(2), 121–125.
Kizhakkayil J., Sasikumar B., 2010. Genetic diversity analysis of ginger (Zingiber officinale Rosc.) germplasm based on RAPD and ISSR markers. Sci. Hortic, 125(1), 73–76.
Li R., Dao Z., 2011. Molecular authentication of the traditional Tibetan medicinal plant, Meconopsis impedita. Afr. J. Biot., 10(38), 7493–7496.
Macedo S.B., Ferreira L.R., Perazzo F.F., Tavares Carvalho J.C., 2004. Anti-inflammatory activity of Arnica Montana 6cH: preclinical study in animals. Homeopathy, 93(2), 84–87.
Nei M., Li W.H., 1979. Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc. Natl. Acad. Sci., 76, 5269–5273.
Okoń S., Surmacz-Magdziak A., 2011. The use of RAPD markers for detecting genetic similarity and molecular identification of chamomile (Chamomilla recutita (L.) Rausch.) genotypes. Herba Pol., 57(1), 38–47.
Okoń S., Surmacz-Magdziak A., Paczos-Grzęda E., 2013. Genetic diversity among cultivated and wild chamomile germplasm based on ISR analysis. Acta Sci. Pol., Hortorum Cultus, 12(2), 43–50.
Pop M., Camelia Sand R., Barbu C.H., 2008. Genetic distance determination in one genotypes of Arnica montana L., by RAPD technique. Bulletin UASVM, Agriculture, 65(1), 1843–5246.
Prakash S., Van Staden J., 2007. Assessment of genetic relationships between Rhus L. species using RAPD markers. Genet. Resour. Crop Ev., 54(1), 7–11.
Prevost A., Wilkinson M.J., 1999. A new system of comparing PCR primers applied to ISSR fingerprinting of potato cultivars. Theor. Appl. Genet., 98(1), 107–112.
Rohlf F.J., 2001. NTSYS-pc numerical taxonomy and multivariate analysis system. Version 5.1. Exeter Publish. Ltd., Setauket, N.Y.
Tan S.C., Yiap B.C., 2009. DNA, RNA and protein extraction: The past and the present. J. Biomed. Biotechnol., 10.
Tatikonda L., Wani S.P., Kannan S., Beerelli N., Sreedevi T.K., Hoisington D.A., Devi P., Varshney R.K., 2009. AFLP-based molecular characterization of an elite germplasm collection of Jatropha curcas L., a biofuel plant. Plant Sci., 176(4), 505–513.
Widrig R., Suter A., Saller R., Melzer J., 2007. Choosing between NSAID and arnica for topical treatment of hand osteoarthritis in a randomised, double-blind study. Rheumat. Internat., 27(6), 585–591.
Williams J.G.K., Kubelik A.R., Livak K.J., Rafalski J.A., Tingey S.V., 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucl. Acid. Res., 18, 6531–6535.
University of Life Science in Lublin
University of Life Science in Lublin
University of Life Science in Lublin
University of Life Science in Lublin
Licencja
Artykuły są udostępniane na zasadach CC BY 4.0 (do 2021 r. na zasadach CC BY-NC-ND 4.0 międzynarodowe).
Przysłanie artykułu do redakcji oznacza, że nie był on opublikowany wcześniej i nie jest rozpatrywany do publikacji gdzie indziej.
Autor podpisuje oświadczenie o oryginalności dzieła, wkładzie poszczególnych osób i źródle finansowania.
