Detection of potyviruses by non-radioactive nucreic acid probe.

点击翻译按钮获取中文摘要

Potyvirus group is one of the biggest group in plant viruses and very important in agriculture. The purpose of this research is to make a specific nonradioactive nucleic acid probe for detecting each polyvirus and its strains. A virus, designated NFU, isolated from bean plants(Phaseolus vulgaris)with severe vine necrosis(Tsurugare-byo)in Fukushima Prefecture was compared with a seed-borne isolate of bean yellow mosaic virus(BYMV-SB)from broad bean. Based on the results of inoculation test, serological test and western blotting, NFU was identified as a strain of clover yellow vein virus(CYVV-NFU). CYVV-RNA was extracted from purified virus preparations and complementary DNA was prepared by the procedure of Gubler and Hoffman. The CDNA was ligated into Smal site of PUC19 plasmid. The E. coli transformed by plasmids containing CDNA inserts were immunoscreened with CYVV antiserum. Three immunopositive clones were obtained. In hybridization test using CDNA probe of CYVV-RNA labeled with photobiotin, a non-radioactive chemical, CYVV was detected very easily and sensitively. Two strains of turnip mosaic virus(TUMV)were also studied. Radish and cabbage strains were distinguished by host range but not by serological method. The CDNA clones were obtained and used as non-radioactive probes.