685 KB81 KB201320252013Bajc, MarkoThaler, Nejcštevilka:100str. 25-40ISSN:2335-3112COBISSID:3636646URN:URN:NBN:SI:doc-0V56L74NslBiotehniška fakulteta, Oddelek za gozdarstvo in obnovljive gozdne vireBiotehniška fakulteta, Oddelek za lesarstvoGozdarski inštitut Slovenije, založba Silvae SlovenicaActa silvae et ligniizolacija DNAPCRpolimerazapolymerase chain reactionpomnoževanje DNAsekundarni metabolitiverižna reakcijaVpliv glivnih in rastlinskih sekundarnih metabolitov na verižno reakcijo s polimerazo (PCR)| Effect of fungal and plant secondary metabolites on polimerase chain reaction (PCR)|Secondary metabolites are organic compounds that can be found in both fungi and plants, where they play an important role as defensive and signal molecules, or provide other kinds of advantage in natural selection, but are not directly involved in normal growth, development and reproduction of an organism. When working with DNA techniques, it is the secondary metabolites that most often affect the efficiency of polymerase chain reaction (PCR), either by hindering cell lysis, causing decomposition of nucleic acids or by direct inhibition of DNA-polymerase or reverse transcriptase amplification. The main limiting factor in the application of the PCR technique in routine diagnosis is the preparation of good quality nucleic acids isolates, free of PCR inhibitors. This is especially true in the case of woody plants (Minafra et al., 1992) and soil samples (Tsai and Olson, 1991). Most standard nucleic acids extraction procedures do not always remove contaminating plant polysaccharides or polyphenolic compounds, which can have direct inhibitory effects on subsequent PCR amplification (Demeke and Adams, 1992). Attempts to overcome these limitations included the development of more elaborate nucleic acids extraction methods and PCR, which employ PCR enhancers to eliminate or attenuate the effects of inhibitors. This review is concentrated on removal or attenuation of effects of plant and fungal secondary metabolites from soil, different plant tissues and decayed wood samples due to the significance of this type of research in forestry and wood scienceSekundarni metaboliti so organske spojine, ki jih najdemo pri glivah in rastlinah, kjer imajo vlogo obrambnih in signalnih molekul ali zagotavljajo druge selekcijske prednosti, niso pa neposredno vpleteni v rast, razvoj in razmnoževanje organizma. Pri delu s tehnikami DNA so pogosto ravno sekundarni metaboliti tisti, ki posredno ali neposredno vplivajo na uspešnost verižne reakcije s polimerazo (PCR) ali reverezno transkriptazo, in sicer tako, da otežujejo celično lizo, povzročajo razpad nukleinskih kislin ali neposredno ovirajo delovanje encima polimeraze pri pomnoževanju tarčne DNK. Glavna ovira pri aplikaciji tehnike PCR v rutinski diagnostiki je priprava visoko kvalitetne DNA brez inhibitorjev. Še posebej to velja pri izolaciji DNA iz lesnatih rastlin (Minafra in sod., 1992) in vzorcev tal (Tsai in Olson, 1991). Večina standardnih postopkov izolacije nukleinskih kislin ne odstrani rastlinskih polisaharidov in polifenolnih komponent, ki imajo lahko neposreden vpliv na pomnoževanje s PCR (Demeke in Adams, 1992). Poskusi za premostitev tovrstnih ovir vključujejo bolj dovršene metode za izolacijo nukleinskih kislin in PCR, ki vključujejo uporabo pospeševalcev PCR za odstranitev ali zmanjšanje vpliva inhibitorjev PCR. Ta pregled je osredotočen na pristope za odstranitev ali zmanjšanje vplivov rastlinskih in glivnih sekundarnih metabolitov iz vzorcev tal, različnih rastlinskih tkiv in razkrojenega lesa zaradi pomena tovrstnih raziskav za gozdarstvo in lesarstvoTEXTznanstveno časopisjejournalsSlovenian National E-content AggregatorNational and University Library of SloveniaGozdarski inštitut Slovenije