An isolate of PVS,HZ00P1,was obtained from nature infected Solanum tuberosum from Hangzhou,Zhejiang province.It was primarily identified by DAS-ELISA and host reaction tests.3’ end partial sequence of the virus isola...An isolate of PVS,HZ00P1,was obtained from nature infected Solanum tuberosum from Hangzhou,Zhejiang province.It was primarily identified by DAS-ELISA and host reaction tests.3’ end partial sequence of the virus isolate was cloned.Nucleic acid sequence of the cp gene and deduced cp amino acid sequence alignments were compared with 16 other isolates registered in GenBank.The results showed that the 17 PVS isolates were divided into two groups.Among them,two Andean isolates were classified to groupⅡ,HZ00P1 and the other 14 isolates were assigned to groupⅠ.Compared with other PVS isolates in group Ⅰ,PVS HZ00P1 had 93.1%-98.1% and 95.9%-99.3% homologies of nucleotide sequence and deduced amino acid sequence respectively,whereas its similarities to the groupⅡwere 81.4%-81.7% and 93.5%-93.9%.A survey of PVS occurrence in Zhejiang province was achieved by RNA spot hybridization(RSH).Among the 30 samples collected from different areas of the province,26.7% were found to have the infection of PVS.It is concluded that PVS has become a common virus in Zhejiang province.展开更多
Potato virus S （PVS） often causes significant losses in potato production in potato-growing countries. In this study, the ordinary strain of PVS （PVS 0） was purified from PVS-infected potato plants and used as the...Potato virus S （PVS） often causes significant losses in potato production in potato-growing countries. In this study, the ordinary strain of PVS （PVS 0） was purified from PVS-infected potato plants and used as the immunogen to produce hybridomas secreting monoclonal antibodies （MAbs）. Five highly specific and sensitive murine MAbs （1A3, 16C10, 18A9, 20B12, and 22H4） against PVS were prepared using conventional hybridoma technology. Using these MAbs, tissue print-enzyme-linked immunosorbent assay （ELISA）, dot-ELISA, and double-antibody sandwich （DAS）- ELISA were developed for sensitive and specific detection of PVS infection in potato plants. The results of sensitivity assays revealed that PVS could be reliably detected in PVS-infected leaf crude extracts diluted at 1：10240 and 1：163840 （w/v, g/ml）in phosphate buffer saline （PBS） by dot-ELISA and DAS-ELISA, respectively. Twenty-two samples collected from potato fields in Yunnan Province, China were tested for PVS infection using the serological assays we had developed, and 14 of them were found to be positive. This indicates that PVS is now prevalent in potato fields in Yunnan Province.展开更多
文摘An isolate of PVS,HZ00P1,was obtained from nature infected Solanum tuberosum from Hangzhou,Zhejiang province.It was primarily identified by DAS-ELISA and host reaction tests.3’ end partial sequence of the virus isolate was cloned.Nucleic acid sequence of the cp gene and deduced cp amino acid sequence alignments were compared with 16 other isolates registered in GenBank.The results showed that the 17 PVS isolates were divided into two groups.Among them,two Andean isolates were classified to groupⅡ,HZ00P1 and the other 14 isolates were assigned to groupⅠ.Compared with other PVS isolates in group Ⅰ,PVS HZ00P1 had 93.1%-98.1% and 95.9%-99.3% homologies of nucleotide sequence and deduced amino acid sequence respectively,whereas its similarities to the groupⅡwere 81.4%-81.7% and 93.5%-93.9%.A survey of PVS occurrence in Zhejiang province was achieved by RNA spot hybridization(RSH).Among the 30 samples collected from different areas of the province,26.7% were found to have the infection of PVS.It is concluded that PVS has become a common virus in Zhejiang province.
基金Project supported by the National Key Research and Development Project of China(No.2017YFD0201604)the Fund for Agroscientific Research in the Public Interest(No.201303028),China
文摘Potato virus S （PVS） often causes significant losses in potato production in potato-growing countries. In this study, the ordinary strain of PVS （PVS 0） was purified from PVS-infected potato plants and used as the immunogen to produce hybridomas secreting monoclonal antibodies （MAbs）. Five highly specific and sensitive murine MAbs （1A3, 16C10, 18A9, 20B12, and 22H4） against PVS were prepared using conventional hybridoma technology. Using these MAbs, tissue print-enzyme-linked immunosorbent assay （ELISA）, dot-ELISA, and double-antibody sandwich （DAS）- ELISA were developed for sensitive and specific detection of PVS infection in potato plants. The results of sensitivity assays revealed that PVS could be reliably detected in PVS-infected leaf crude extracts diluted at 1：10240 and 1：163840 （w/v, g/ml）in phosphate buffer saline （PBS） by dot-ELISA and DAS-ELISA, respectively. Twenty-two samples collected from potato fields in Yunnan Province, China were tested for PVS infection using the serological assays we had developed, and 14 of them were found to be positive. This indicates that PVS is now prevalent in potato fields in Yunnan Province.
