Analysis of ISEM, SDS PAGE, Western blot, Northern blot and RT PCR indicated that the soil borne mosaic disease occurred on winter wheat in Yantai district, Shandong Province, was caused by co infection of two differe...Analysis of ISEM, SDS PAGE, Western blot, Northern blot and RT PCR indicated that the soil borne mosaic disease occurred on winter wheat in Yantai district, Shandong Province, was caused by co infection of two different viruses. The filamentous virus was identified to be wheat yellow mosaic virus (WYMV) as it had a coat protein of 33kD, which was serologically related to WYMV, wheat spindle streak mosaic virus(WSSMV) and barley yellow mosaic virus (BaYMV), but not to barley mild mosaic virus(BaMMV) and soil borne wheat mosaic virus(SBWMV). In addition, DNA fragments specifically amplified from its cDNAs template with WYMV primers were identical to those amplified from the homologous template. The rod shaped virus was identified to be a SBWMV related virus as it had a coat protein of 19 kD, which was serologically related to SBWMV and TMV, but not to WYMV, WSSMV, BaYMV, BaMMV and three monoclonal antibodies to SBWMV. Its RNA1 (7.1kb) and RNA2 (3.6 kb) were respectively hybridized by cDNA probes of RNA1(p3H5) and RNA2(pJC2G6) of SBWMV, but no DNA fragments specifically amplified from cDNA template of its RNA with primers of SBWMV RNA 1 and RNA2.展开更多
Based on multi-alignment of complete polyprotein amino acid sequences of genus Potyvirus,five degenerated primers were designedThey were Sprimer(5′-GGX AAY AAY AGY GGX CAZ CC-3′),pNIa(+)(5′-TNY TGG AAM CAY TGG A...Based on multi-alignment of complete polyprotein amino acid sequences of genus Potyvirus,five degenerated primers were designedThey were Sprimer(5′-GGX AAY AAY AGY GGX CAZ CC-3′),pNIa(+)(5′-TNY TGG AAM CAY TGG AT-3′),pCI2(+)(5′-GCX ACX AAX ATX ATX GAX AA-3′),pCI1(+)(5′-GTX GGX TCX GGX AAX TCX AC-3′)and pHC(+)(5′-TGY GAY AAY CAZ TTX GA-3′)(X=A,T,C or G:Y=T or C;Z=A or G;N=A or T;M=A,T or G)Using degenerated PCR and modified RACE methods,a protocol for determination of complete genome sequence of potyviruses was established and proved to be successful on five展开更多
基金欧共体资助项目(INCO IC 18-C796-0049)863启动项目+2 种基金中华农业科教基金(97-03-B-6)国家(39600098)和浙江省自然科学基金(396029)浙江省青年科技人才资金(RC9604)
文摘Analysis of ISEM, SDS PAGE, Western blot, Northern blot and RT PCR indicated that the soil borne mosaic disease occurred on winter wheat in Yantai district, Shandong Province, was caused by co infection of two different viruses. The filamentous virus was identified to be wheat yellow mosaic virus (WYMV) as it had a coat protein of 33kD, which was serologically related to WYMV, wheat spindle streak mosaic virus(WSSMV) and barley yellow mosaic virus (BaYMV), but not to barley mild mosaic virus(BaMMV) and soil borne wheat mosaic virus(SBWMV). In addition, DNA fragments specifically amplified from its cDNAs template with WYMV primers were identical to those amplified from the homologous template. The rod shaped virus was identified to be a SBWMV related virus as it had a coat protein of 19 kD, which was serologically related to SBWMV and TMV, but not to WYMV, WSSMV, BaYMV, BaMMV and three monoclonal antibodies to SBWMV. Its RNA1 (7.1kb) and RNA2 (3.6 kb) were respectively hybridized by cDNA probes of RNA1(p3H5) and RNA2(pJC2G6) of SBWMV, but no DNA fragments specifically amplified from cDNA template of its RNA with primers of SBWMV RNA 1 and RNA2.
文摘Based on multi-alignment of complete polyprotein amino acid sequences of genus Potyvirus,five degenerated primers were designedThey were Sprimer(5′-GGX AAY AAY AGY GGX CAZ CC-3′),pNIa(+)(5′-TNY TGG AAM CAY TGG AT-3′),pCI2(+)(5′-GCX ACX AAX ATX ATX GAX AA-3′),pCI1(+)(5′-GTX GGX TCX GGX AAX TCX AC-3′)and pHC(+)(5′-TGY GAY AAY CAZ TTX GA-3′)(X=A,T,C or G:Y=T or C;Z=A or G;N=A or T;M=A,T or G)Using degenerated PCR and modified RACE methods,a protocol for determination of complete genome sequence of potyviruses was established and proved to be successful on five
