Studies On Chloroplast Dna Restriction Fragment Length Polymorphisms Of Three Species Of Guizotia Casso (g. Abyssinicag.scabra Ssp. Scabra And G.scabra Ssp. Schimperi)

Chloroplast DNA from three Guizotia species (G. abyssinica, G.rnscabra ssp. scabra and G. scabra ssp. schimperi) were isolatedrnusing sucrose gradient centrifugation method and Dynabeads DNArnDIRECTTH System I. Each of the DNA extracted by sucroserngradient method was separetely digested with BamHI, EcoRI andrnHindIII restriction endonuclease enzymes, recognizing 6 basernfor 4hr. The digestion products were thenrnloaded on 0.8% agarose gel and electrophoresed at 30-50 voltsrnfor 3-4 hours. After visualizing with o. 5l1g/ml of ethidiumrnbromide solution and photographed using polaroid films, thernfragment bands were compared. In addition, the DNAs extractedrnby Dynabeads DNA DIRECTT1rn-{ System I method were used for PCRrnamplification of chloroplast DNA non-coding region (trnL (UAA)rnintron) using Primer C and D of Taberlet et al. (1991). Thernamplified chloroplast DNA non-coding "region, I-las digested withrnSau3AI restriction endonuclease enzyme recognizing 4 basernpairs at 37°C for 2 hr. and electrophoresed in 2% agarose gelrnat 100 volts for 2 hr. After being visualized with 0.5I1g/mlrnethidium bromide solution and photographed using polaroidrnfilms, the fragment bands I~ere compared. In both cases,rndetectable variation I~as not observed among the Guizotiarnspecies compared, indicating that these species are eitherrnclosely related and recently diversified or their chloroplastrnDNAs are evolving at a relatively slO~er rates in comparisonrnto the rates of cpDNA evolution in other angiosperm species.

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