MOLECULAR-CLONING AND DETECTION OF DNA OF THE MYCOPLASMALIKE ORGANISM ASSOCIATED WITH CLOVER PROLIFERATION

被引：21

作者：

DENG, SJ

HIRUKI, C

机构：

[1] Department of Plant Science, University of Alberta, Edmonton, AB

来源：

CANADIAN JOURNAL OF PLANT PATHOLOGY | 1990年 / 12卷 / 04期

基金：

加拿大自然科学与工程研究理事会;

关键词：

D O I：

10.1080/07060669009500978

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

A fraction enriched with mycoplasmalike organism (MLO) was prepared from midrib and petiole tissues of periwinkle (Catharanthus roseus) plants affected by clover proliferation (CP). DNA isolated from the MLO-enriched extract was digested with both EcoRI and HindIII restriction endonucleases, ligated with plasmid pUC19, and cloned in E. coli DH5a. A recombinant plasmid containing 2.99 kb of CP MLO DNA segment was either radioactively or biotinylate labeled. The labeled cloned DNA probe hybridized with DNA isolated from CP MLO-infected periwinkle plants but not with DNA extracted from healthy periwinkle plants. At least 2 ng of total nucleic acids from periwinkle plants are needed to detect CP MLO with biotinylated DNA probe. Dot-blot hybridization detected CP MLO DNA 2 weeks in advance of symptom development and showed that CP MLO reached the highest concentration in the periwinkle plants 7 weeks after graft-transmission. The probe also cross- hybridized to nucleic acids from potato witches'-broom (PWB) MLO-infected periwinkle but did not cross-hybridize to nucleic acids from western aster yellows (AY-27) MLO-infected periwinkle or clover phyllody (CPD) MLO-infected clover.

引用

页码：383 / 388

页数：6

共 29 条

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