Studies on Chloroplast Dna Restriction Fragment Length Polymorphisms of Three Species of Guizotia Casso (G. Abyssinica,G.Scabra Ssp. Scabra and G.Scabra Ssp. Schimperi)
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Date
1999-05
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Addis Ababa University
Abstract
Chloroplast DNA from three Guizotia species (G. abyssinica, G.
scabra ssp. scabra and G. scabra ssp. schimperi) were isolated
using sucrose gradient centrifugation method and Dynabeads DNA
DIRECTTH System I. Each of the DNA extracted by sucrose
gradient method was separetely digested with BamHI, EcoRI and
HindIII restriction endonuclease enzymes, recognizing 6 base
for 4hr. The digestion products were then
loaded on 0.8% agarose gel and electrophoresed at 30-50 volts
for 3-4 hours. After visualizing with o. 5l1g/ml of ethidium
bromide solution and photographed using polaroid films, the
fragment bands were compared. In addition, the DNAs extracted
by Dynabeads DNA DIRECTT1
-{ System I method were used for PCR
amplification of chloroplast DNA non-coding region (trnL (UAA)
intron) using Primer C and D of Taberlet et al. (1991). The
amplified chloroplast DNA non-coding "region, I-las digested with
Sau3AI restriction endonuclease enzyme recognizing 4 base
pairs at 37°C for 2 hr. and electrophoresed in 2% agarose gel
at 100 volts for 2 hr. After being visualized with 0.5I1g/ml
ethidium bromide solution and photographed using polaroid
films, the fragment bands I~ere compared. In both cases,
detectable variation I~as not observed among the Guizotia
species compared, indicating that these species are either
closely related and recently diversified or their chloroplast
DNAs are evolving at a relatively slO\~er rates in comparison
to the rates of cpDNA evolution in other angiosperm species.
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Biology