ISOLATION OF THE DNA OF VARIOUS PLANT PATHOGENIC MYCOPLASMALIKE ORGANISMS FROM INFECTED PLANTS
ISOLATION OF THE DNA OF VARIOUS PLANT PATHOGENIC MYCOPLASMALIKE ORGANISMS FROM INFECTED PLANTS
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DOI：
10.1094/phyto-80-233
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发表时间：
1990-03-01
期刊：
影响因子：
3.2
通讯作者：
BOVE, JM
中科院分区：
文献类型：
--
作者：
KOLLAR, A;SEEMULLER, E;BOVE, JM
用两种不同的方法分别从苹果增殖病的长春花植株和苹果树以及引起油菜绿变、长春花绿变和双斑叶蝉叶状体的类支原体感染的长春花中提取DNA。重复的双苯甲酰亚胺-氯化铯浮力密度梯度离心用于分离MLO DNA从宿主植物DNA。由于其低G + C含量，MLO DNA作为显示出比宿主植物DNA低的浮力密度的良好分辨的条带获得。该带高度富集MLO DNA，占从患病植物提取的总DNA的0.1-3.0%。将苹果增殖剂的DNA克隆到大肠杆菌中，并通过斑点杂交和Southern印迹杂交证实了克隆的MLO DNA的特异性。
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.