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
Full-length RNA4 of beet necrotic yellow vein virus (BNYVV) and its mutants, including a frame-shift and deletions within the coding region, were inserted into a transcription plasmid. Transcripts derived from these p...Full-length RNA4 of beet necrotic yellow vein virus (BNYVV) and its mutants, including a frame-shift and deletions within the coding region, were inserted into a transcription plasmid. Transcripts derived from these plasmids and RNAs extracted from a BNYVV isolate containing only RNAs 1, 2 and 3 were coinoculated to Tetragona expansa. Then these recombinant isolates were inoculated to sugar beet plants grown in a sand culture system containing Po-lymyxa betae that had been freed of virus particles to test efficiency of BNYVV transmission. The result showed that the efficiency of BNYVV transmission among the plants by Polymyxa betae was highly decreased by different deletions in the coding region of RNA4, but not affected by the frame-shift mutant with four nucleotides insertion. The observations suggest that at least some of the RNA4 sequence encompassing the deleted region of 580 nucleotide acids (525 -1105 nt) are necessary for high efficient transmission of the virus by the fungus.展开更多
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.
基金This work was partially supported by the National Natural Science Foundation of China (Grant No. 39670035)the State "863" High-Tech Program Specifical Fund for Young Scientists.
文摘Full-length RNA4 of beet necrotic yellow vein virus (BNYVV) and its mutants, including a frame-shift and deletions within the coding region, were inserted into a transcription plasmid. Transcripts derived from these plasmids and RNAs extracted from a BNYVV isolate containing only RNAs 1, 2 and 3 were coinoculated to Tetragona expansa. Then these recombinant isolates were inoculated to sugar beet plants grown in a sand culture system containing Po-lymyxa betae that had been freed of virus particles to test efficiency of BNYVV transmission. The result showed that the efficiency of BNYVV transmission among the plants by Polymyxa betae was highly decreased by different deletions in the coding region of RNA4, but not affected by the frame-shift mutant with four nucleotides insertion. The observations suggest that at least some of the RNA4 sequence encompassing the deleted region of 580 nucleotide acids (525 -1105 nt) are necessary for high efficient transmission of the virus by the fungus.
