In fission yeast (Schizosaccharomyces pombe), pac1 gene was cloned with 99.3% nucleo- tide sequence similarity with published pac1 in GenBank. In pET-5α expression system, the expres- sion product of cloned pac1 in E...In fission yeast (Schizosaccharomyces pombe), pac1 gene was cloned with 99.3% nucleo- tide sequence similarity with published pac1 in GenBank. In pET-5α expression system, the expres- sion product of cloned pac1 in E. coli showed activity to degrade the double-strand RNA. Harboring the binary vector pBI121, which contains pac1 gene, Agrobacterium tumefaciens strain LBA4404 was used to transform the wheat immature embryos pre- cultured 7―10 d. After preregeneration, regeneration and selection culture stage, totally 41 G418 resistant plants were obtained, in which 25 lines were proved to integrate with transgene and express transgene normally by PCR, Dot blot, RT-PCR and ELISA de- tection. Antivirus test carried out on 25 positive lines with high dose of Barley yellow dwarf virus-GPV re- vealed that 12 lines had resistance to BVDV-GPV in low level, another 12 lines had resistance to BVDV- GPV in middle level, and 1 line showed resistance to BVDV-GPV in high level. However, both low and middle level of resistance plants showed no symp- toms when infected by viruses at low dose, which suggested the dose-dependent effect of the resis- tance mediated by pac1 to BYDV-GPV.展开更多
Barley yellow dwarf virus (BYDV) is one of the most serious wheat diseases in China. So far no resistance has been described in common wheat. A certain level of BYDV resistance was found in thirteen Triticeae species....Barley yellow dwarf virus (BYDV) is one of the most serious wheat diseases in China. So far no resistance has been described in common wheat. A certain level of BYDV resistance was found in thirteen Triticeae species. Thinopyrum intermedium, two octoploids derived from TH. intermedium/wheat, Zhong 4 awnless and TAF46, and one disomic addition line, L1 derived from TAF46, showed good resistance to BYDV by enzyme linked immunosorbent assay (ELISA). Two wheat/TA. intermedium translocation lines, CPI 119880 and CPI 119899, showing good BYDV resistance were developed from L1 by using both CSph mutant and tissue culture. It is found that their BYDV resistance was controlled by a single dominant gene. Two cDNA probes pEleAcc3 and pPJN8 (E1-T1) were screened for detecting Th. intermedium DNA in wheat background. A specific band for the DNA of Th. intermedium and its derivatives was found in Southern hybridization. It is also possible to determine the size of the alien segment by comparing the relative density of the specific band. Therefore, this can be used as a marker to identify the BYDV resistance in wheat breeding program.展开更多
文摘In fission yeast (Schizosaccharomyces pombe), pac1 gene was cloned with 99.3% nucleo- tide sequence similarity with published pac1 in GenBank. In pET-5α expression system, the expres- sion product of cloned pac1 in E. coli showed activity to degrade the double-strand RNA. Harboring the binary vector pBI121, which contains pac1 gene, Agrobacterium tumefaciens strain LBA4404 was used to transform the wheat immature embryos pre- cultured 7―10 d. After preregeneration, regeneration and selection culture stage, totally 41 G418 resistant plants were obtained, in which 25 lines were proved to integrate with transgene and express transgene normally by PCR, Dot blot, RT-PCR and ELISA de- tection. Antivirus test carried out on 25 positive lines with high dose of Barley yellow dwarf virus-GPV re- vealed that 12 lines had resistance to BVDV-GPV in low level, another 12 lines had resistance to BVDV- GPV in middle level, and 1 line showed resistance to BVDV-GPV in high level. However, both low and middle level of resistance plants showed no symp- toms when infected by viruses at low dose, which suggested the dose-dependent effect of the resis- tance mediated by pac1 to BYDV-GPV.
基金This research is supported by the Australian Centre for International Agricultural Research(Projects 8379 and 8813)by the National Science and Technology Committee of China on China's side.
文摘Barley yellow dwarf virus (BYDV) is one of the most serious wheat diseases in China. So far no resistance has been described in common wheat. A certain level of BYDV resistance was found in thirteen Triticeae species. Thinopyrum intermedium, two octoploids derived from TH. intermedium/wheat, Zhong 4 awnless and TAF46, and one disomic addition line, L1 derived from TAF46, showed good resistance to BYDV by enzyme linked immunosorbent assay (ELISA). Two wheat/TA. intermedium translocation lines, CPI 119880 and CPI 119899, showing good BYDV resistance were developed from L1 by using both CSph mutant and tissue culture. It is found that their BYDV resistance was controlled by a single dominant gene. Two cDNA probes pEleAcc3 and pPJN8 (E1-T1) were screened for detecting Th. intermedium DNA in wheat background. A specific band for the DNA of Th. intermedium and its derivatives was found in Southern hybridization. It is also possible to determine the size of the alien segment by comparing the relative density of the specific band. Therefore, this can be used as a marker to identify the BYDV resistance in wheat breeding program.
