Isolation of the DNA of Various Plant Pathogenic Mycoplasmalike Organisms from Infected Plants

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Abstract
DNA from apple proliferation-diseased periwinkle plants and apple trees and from periwinkles infected with mycoplasmalike organisms (MLOs) causing a virescence (chloranty) of rape, a virescence of periwinkle, and a phyllody of Diplotaxis erucoides was extracted by using two different methods. Repeated bisbenzimide-CsCl buoyant density gradient centrifugations were used to separate MLO DNA from host plant DNA. MLO DNA was obtained as a well-resolved band showing a lower buoyant density than the host plant DNA due to its low G + C content. This band was highly enriched in MLO DNA and comprised 0.1-3.0% of the total DNA extracted from diseased plants. The DNA of the apple proliferation agent was cloned in Escherichia coli and the specificity of the cloned MLO DNA was confirmed by dot and Southern blot hybridization.