Wheat yellow mosaic is a soil-borne virus disease,and now has become one of the main viral diseases endangering wheat production in China,posing serious threat to wheat production. The pathogen,transmission media,inci...Wheat yellow mosaic is a soil-borne virus disease,and now has become one of the main viral diseases endangering wheat production in China,posing serious threat to wheat production. The pathogen,transmission media,incidence regularity and performance of wheat yellow mosaic were introduced in the paper,and its molecular biology and breeding research were summarized,in order to provide a reference for resistance breeding and production practice of wheat yellow mosaic.展开更多
Wheat seeds of 109 cultivars from USA, Europe and Japan were sown in experiments at seven sites in different provinces of China for one or two seasons. Five of the sites were infested with the bymovirus wheat yellow m...Wheat seeds of 109 cultivars from USA, Europe and Japan were sown in experiments at seven sites in different provinces of China for one or two seasons. Five of the sites were infested with the bymovirus wheat yellow mosaic virus (WYMV) and two jointly with WYMV and the furovirus Chinese wheat mosaic virus (CWMV). Disease symptoms were assessed visually and leaf samples were tested for virus (es) by ELISA. At least 29 cultivars were resistant to WYMV at the sites where only this virus was present but all the cultivars were severely infected at Rongcheng (Shandong Province) where CWMV was mixed with WYMV. There was evidence that the presence of CWMV assisted infection by WYMV and also resulted in more severe symptoms. At the mixed site in Yantai, Shandong Province, symptoms were mild and many cultivars had symptomless infection . Of the two strains of WYMV identified in Japan, the Chinese sites seem to be most similar to the type isolated, WYMV-T. Eleven cultivars seemed to be susceptible to WYMV only at Loutian (Hubei Province),suggesting that the virus at this site would be worth studying further.展开更多
Wheat yellow mosaic virus (WYMV) isolate HC was used for viral cDNA synthesis and sequencing. The results show that the viral RNA1 is 7 629 nucleotides encoding a polyprotein with 2 407 amino acids, from which seven p...Wheat yellow mosaic virus (WYMV) isolate HC was used for viral cDNA synthesis and sequencing. The results show that the viral RNA1 is 7 629 nucleotides encoding a polyprotein with 2 407 amino acids, from which seven putative proteins may be produced by an autolytic cleavage processing besides the viral coat protein. The RNA2 is 3 639 nucleotides and codes for a polyprotein of 903 amino acids, which may contain two putative non-structural proteins. Although WYMV shares a similarity in genetic organization to wheat spindle streak mosaic virus (WSSMV), the identities in their nucleotide sequences or deduced amino acid sequences are as low as 70% and 75% respectively. Based on this result, it is confirmed that WYMV and WSSMV are different species within Bymovirus .展开更多
The plasmid containing the promoter Actl, the coat protein (cp) gene of wheat yellow mosaic virus (WYMV) and the selectable bar gene, was delivered via particle bombardment, directly into immature embryos of a wheat c...The plasmid containing the promoter Actl, the coat protein (cp) gene of wheat yellow mosaic virus (WYMV) and the selectable bar gene, was delivered via particle bombardment, directly into immature embryos of a wheat culti-vars. PCR and PCR-RFLP were employed to screen the existence of the cp gene in T0 and T1 generations. Seeds from the positive T1 plants were sowed in fields heavily contaminated with WYMV to detect their resistance. In field trial of virus infection, one of the transgenic wheat lines, P8-T2, exhibited highly disease-resistance. Western blot and RT-PCR analysis showed that the expression level of cp gene in the resistant transgenic line was reduced greatly compared to those susceptible to WYMV infection. This provided evidence to presume that the resistance obtained by the transgenic wheat line was stimulated by the mechanism of the virus induced gene silencing.展开更多
By reverse transcription-polymerase chain reaction (RT-PCR), cDNA fragment of wheat yellow mosaic vir黶 (WYMV) RNA2 encoding 72 ku protein has been synthesized and cloned into plasmid pET30a(+) for overexpression in p...By reverse transcription-polymerase chain reaction (RT-PCR), cDNA fragment of wheat yellow mosaic vir黶 (WYMV) RNA2 encoding 72 ku protein has been synthesized and cloned into plasmid pET30a(+) for overexpression in prokaryotic celis. BL21(DE3) pLys S of E.coli transformed with the recombinant plasmid pETP72 containing the fragment has been induced to express the 72 ku protein on high level. The produced protein has been purified from sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) for its antiserum preparation. in western-blotting analysis, the antibodies reacted with the 72 ku protein expressed in E.coli.展开更多
基金Supported by Major Technology Project of Independent Innovation in Shandong Province ( 2014GJJS0201)Major Scientific and Technological Achievements Cultivation Project of Shandong Academy of Agricultural Sciences( 2014CGPY03)+2 种基金National Key Technology R&D Program ( 2014BAA03B042013BAD01B02)Special Fund for National Scientific Research in the Public Interest ( 201303007-1)
文摘Wheat yellow mosaic is a soil-borne virus disease,and now has become one of the main viral diseases endangering wheat production in China,posing serious threat to wheat production. The pathogen,transmission media,incidence regularity and performance of wheat yellow mosaic were introduced in the paper,and its molecular biology and breeding research were summarized,in order to provide a reference for resistance breeding and production practice of wheat yellow mosaic.
基金the Zhejiang Foun-dation of Natural Sciences(RC9604)National Natural Science Foundation of China(39970482)+3 种基金the Commission of European Union(INCO C118 CT96-0049)a special grant for Zhejiang Provincial KeyLaboratory(001107557) Zhejiang Provincial KeyResearch Programme(2001-2005)(011102181)I-ACR receives grant-aided support from the Biotech-nology and Biological Sciences Research Council of the United Kingdom.
文摘Wheat seeds of 109 cultivars from USA, Europe and Japan were sown in experiments at seven sites in different provinces of China for one or two seasons. Five of the sites were infested with the bymovirus wheat yellow mosaic virus (WYMV) and two jointly with WYMV and the furovirus Chinese wheat mosaic virus (CWMV). Disease symptoms were assessed visually and leaf samples were tested for virus (es) by ELISA. At least 29 cultivars were resistant to WYMV at the sites where only this virus was present but all the cultivars were severely infected at Rongcheng (Shandong Province) where CWMV was mixed with WYMV. There was evidence that the presence of CWMV assisted infection by WYMV and also resulted in more severe symptoms. At the mixed site in Yantai, Shandong Province, symptoms were mild and many cultivars had symptomless infection . Of the two strains of WYMV identified in Japan, the Chinese sites seem to be most similar to the type isolated, WYMV-T. Eleven cultivars seemed to be susceptible to WYMV only at Loutian (Hubei Province),suggesting that the virus at this site would be worth studying further.
基金Project supported by the National Natural Science Foundation of China (Grant No. 39470481)High-Tech. Program of China. The nucleotide sequence data reported in this paper have been submitted to DDBJ/GenBank/EMBL database with the accession number AF0
文摘Wheat yellow mosaic virus (WYMV) isolate HC was used for viral cDNA synthesis and sequencing. The results show that the viral RNA1 is 7 629 nucleotides encoding a polyprotein with 2 407 amino acids, from which seven putative proteins may be produced by an autolytic cleavage processing besides the viral coat protein. The RNA2 is 3 639 nucleotides and codes for a polyprotein of 903 amino acids, which may contain two putative non-structural proteins. Although WYMV shares a similarity in genetic organization to wheat spindle streak mosaic virus (WSSMV), the identities in their nucleotide sequences or deduced amino acid sequences are as low as 70% and 75% respectively. Based on this result, it is confirmed that WYMV and WSSMV are different species within Bymovirus .
基金The field trial was approved by the Committee of Bio-safety Regulation on transgenic Organismsthe Ministry of Agriculture, China (Grant No. 2000B-01-053)+1 种基金This work was supported jointly by the State High-Tech Program (Grant No. 101-04-01-05)the Nati
文摘The plasmid containing the promoter Actl, the coat protein (cp) gene of wheat yellow mosaic virus (WYMV) and the selectable bar gene, was delivered via particle bombardment, directly into immature embryos of a wheat culti-vars. PCR and PCR-RFLP were employed to screen the existence of the cp gene in T0 and T1 generations. Seeds from the positive T1 plants were sowed in fields heavily contaminated with WYMV to detect their resistance. In field trial of virus infection, one of the transgenic wheat lines, P8-T2, exhibited highly disease-resistance. Western blot and RT-PCR analysis showed that the expression level of cp gene in the resistant transgenic line was reduced greatly compared to those susceptible to WYMV infection. This provided evidence to presume that the resistance obtained by the transgenic wheat line was stimulated by the mechanism of the virus induced gene silencing.
文摘By reverse transcription-polymerase chain reaction (RT-PCR), cDNA fragment of wheat yellow mosaic vir黶 (WYMV) RNA2 encoding 72 ku protein has been synthesized and cloned into plasmid pET30a(+) for overexpression in prokaryotic celis. BL21(DE3) pLys S of E.coli transformed with the recombinant plasmid pETP72 containing the fragment has been induced to express the 72 ku protein on high level. The produced protein has been purified from sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) for its antiserum preparation. in western-blotting analysis, the antibodies reacted with the 72 ku protein expressed in E.coli.
