The barley yellow dwarf virus(BYDV)resistance lines of Z1 and Z2 were derived from Zhong 5, a partial amphiploid resulted from the cross between Triticum aestivum (wheat) and Thinopyrum intermedium . Genomic in situ h...The barley yellow dwarf virus(BYDV)resistance lines of Z1 and Z2 were derived from Zhong 5, a partial amphiploid resulted from the cross between Triticum aestivum (wheat) and Thinopyrum intermedium . Genomic in situ hybridization (GISH) was used to analyze the chromosome constitution of Zhong 5 by using genomic DNA of Pseudoregneria strigosa (StSt,2 n =14)as the probe. The GISH results showed that zhong 5 contains 42 wheat chromosomes and l4 Th.intermedium chromosomes composed of 4 St, 4 Js,4 St J translocation and 2 St Js Robertsonian translocation chromosomes. The chromosome constitution of Z1 and Z2 was analyzed by GISH using genomic DNA probes from Th.intermedium and Ps.Strigosa . The GISH results indicated that both Z1 and Z2 possess 42 wheat chromosomes and 2 Th.intermedium chromosomes that were identical to a pair of St J translocation chromosomes in Zhong 5. The Th.intermedium chromosomes,designated as 2Ai 2 chromosome derived from Zhong 5,mostly belong to the St genome except the middle region (about one third of the long arm) belonging to the E(J)genome. A detailed RFLP analysis was conducted for Z1,Z2 and their parents,St and E (J) genomes. The results of RFLP analyses demonstrated that the Th.intermedium chromosomes(2Ai 2,St J)in Z1 and Z2 are extensively homologous to the Wheat group 2 chromosomes. The results of RFLP analyses on the genome composition of the 2Ai 2 chromosome were in agreement with the GISH results. Presence of psr 928 on 2AS and 2DS but absence on 2Ai 2S suggests some internal structural differences between 2Ai 2 and the wheat group 2 chromosomes. Some RFLP markers specific to the 2Ai 2 chromosome were identified and may be effectively used to select translocation lines with small segment of the 2Ai 2 chromosome and to localize the BYDV resistance gene in wheat background.展开更多
The complete nucleotide sequence of genomic RNA of BYDV-GAV was determined. It comprised 5685 nucleotides and contained six open reading frames and four un-translated regions. The size and organization of BYDV-GAV gen...The complete nucleotide sequence of genomic RNA of BYDV-GAV was determined. It comprised 5685 nucleotides and contained six open reading frames and four un-translated regions. The size and organization of BYDV-GAV genome were similar to those of BYDV PAV-aus. The nucleotide and deduced amino acid sequences of the six ORFs were aligned and compared with those of other luteoviruses. The results showed that there was a high degree of identity between BYDV-GAV and MAV-PS1 in all ORFs except ORF5 and ORF6, which had only 87.4% and 70.2% identities respectively. The reported genomic nucleotide sequence of MAV was shorter than that of BYDV-GAV, but the comparison of the genomic nucleotide sequences for MAV-PS1 and GAV showed 90.4% sequence identity for the same region of the genome. Ac-cording to the level of sequence similarities, BYDV-GAV should be closely related to BYDV-MAV.展开更多
Using 2-D electrophoresis and virus overlay assay, a 50-kDa protein (P50) exhibiting specific binding to purified virus particles of BYDV-GAV was found in the protein extracts from Schizaphis graminum and Sitobion ave...Using 2-D electrophoresis and virus overlay assay, a 50-kDa protein (P50) exhibiting specific binding to purified virus particles of BYDV-GAV was found in the protein extracts from Schizaphis graminum and Sitobion avenae, two aphid species transmitting BYDV-GAV. P50 in the extracts of S. graminum was isolated by preparation electrophoresis and electro-eluted proteins from the gel slices for antiserum preparation. After feeding the antiserum through membrane, the transmission efficiencies of S. graminum and S. avenae for BYDV-GAV decreased significantly. It was suggested that P50 should be related with transmission pro- cess. Location of P50 was found at the plasma membrane surrounding the accessory salivary gland (ASG) in the head tissues of S. graminum by immunogold-labelling experiment. The ascertainment of the protein associated with virus transmission has a significance influence on further understanding the transmission mechanism and genetic engineering for resistant to vector transmission.展开更多
文摘The barley yellow dwarf virus(BYDV)resistance lines of Z1 and Z2 were derived from Zhong 5, a partial amphiploid resulted from the cross between Triticum aestivum (wheat) and Thinopyrum intermedium . Genomic in situ hybridization (GISH) was used to analyze the chromosome constitution of Zhong 5 by using genomic DNA of Pseudoregneria strigosa (StSt,2 n =14)as the probe. The GISH results showed that zhong 5 contains 42 wheat chromosomes and l4 Th.intermedium chromosomes composed of 4 St, 4 Js,4 St J translocation and 2 St Js Robertsonian translocation chromosomes. The chromosome constitution of Z1 and Z2 was analyzed by GISH using genomic DNA probes from Th.intermedium and Ps.Strigosa . The GISH results indicated that both Z1 and Z2 possess 42 wheat chromosomes and 2 Th.intermedium chromosomes that were identical to a pair of St J translocation chromosomes in Zhong 5. The Th.intermedium chromosomes,designated as 2Ai 2 chromosome derived from Zhong 5,mostly belong to the St genome except the middle region (about one third of the long arm) belonging to the E(J)genome. A detailed RFLP analysis was conducted for Z1,Z2 and their parents,St and E (J) genomes. The results of RFLP analyses demonstrated that the Th.intermedium chromosomes(2Ai 2,St J)in Z1 and Z2 are extensively homologous to the Wheat group 2 chromosomes. The results of RFLP analyses on the genome composition of the 2Ai 2 chromosome were in agreement with the GISH results. Presence of psr 928 on 2AS and 2DS but absence on 2Ai 2S suggests some internal structural differences between 2Ai 2 and the wheat group 2 chromosomes. Some RFLP markers specific to the 2Ai 2 chromosome were identified and may be effectively used to select translocation lines with small segment of the 2Ai 2 chromosome and to localize the BYDV resistance gene in wheat background.
基金This work was supported by the National Key Basic Research of China(973 contract TG2000016201)the National Natural Science Foundation of China(Grant No.39970034).
文摘The complete nucleotide sequence of genomic RNA of BYDV-GAV was determined. It comprised 5685 nucleotides and contained six open reading frames and four un-translated regions. The size and organization of BYDV-GAV genome were similar to those of BYDV PAV-aus. The nucleotide and deduced amino acid sequences of the six ORFs were aligned and compared with those of other luteoviruses. The results showed that there was a high degree of identity between BYDV-GAV and MAV-PS1 in all ORFs except ORF5 and ORF6, which had only 87.4% and 70.2% identities respectively. The reported genomic nucleotide sequence of MAV was shorter than that of BYDV-GAV, but the comparison of the genomic nucleotide sequences for MAV-PS1 and GAV showed 90.4% sequence identity for the same region of the genome. Ac-cording to the level of sequence similarities, BYDV-GAV should be closely related to BYDV-MAV.
基金This work was supported by the National Key Basic Research of China(Grant No.TG2000016201)the National Natural Science Foundation of China(Grant No.30070498).
文摘Using 2-D electrophoresis and virus overlay assay, a 50-kDa protein (P50) exhibiting specific binding to purified virus particles of BYDV-GAV was found in the protein extracts from Schizaphis graminum and Sitobion avenae, two aphid species transmitting BYDV-GAV. P50 in the extracts of S. graminum was isolated by preparation electrophoresis and electro-eluted proteins from the gel slices for antiserum preparation. After feeding the antiserum through membrane, the transmission efficiencies of S. graminum and S. avenae for BYDV-GAV decreased significantly. It was suggested that P50 should be related with transmission pro- cess. Location of P50 was found at the plasma membrane surrounding the accessory salivary gland (ASG) in the head tissues of S. graminum by immunogold-labelling experiment. The ascertainment of the protein associated with virus transmission has a significance influence on further understanding the transmission mechanism and genetic engineering for resistant to vector transmission.
