The gene Pi-d2, conferring gene-for-gene resistance to the Chinese blast strain ZB15, was isolated from a rice variety （Digu） by the map-based cloning strategy. Here, we constructed a control plasmid pZH01-pi-d2tp3...The gene Pi-d2, conferring gene-for-gene resistance to the Chinese blast strain ZB15, was isolated from a rice variety （Digu） by the map-based cloning strategy. Here, we constructed a control plasmid pZH01-pi-d2tp309 （pZH01-tp309） and three different expression constructs, pCB-Pi-d25.3kb （pCB5.3kb）, pCB-Pi-d26.3kb （pCB6.3kb） and pZH01-Pi-d22.72kb （pZH01-2.72kb） of Pi-d2, driven by Pi-d2 gene’s own promoter or CaMV35S promoter. These constructs were separately introduced into japonica rice varieties Lijiangxintuanhegu, Taipei 309, Nipponbare and Zhonghua 9 through Agrobacterium- mediated transformation. A total of 150 transgenic rice plants were obtained from the regenerated calli selected on hygromycin. PCR, RT-PCR and Southern-blotting assay showed that the gene of interest had been integrated into rice genome and stably inherited. Thirty-five transgenic lines independently derived from T1 progeny were inoculated with the rice blast strain ZB15. Transformants exhibited resistance to rice blast at various levels. The lesions on the transgenic plant leaves were less severe than those on the controls and the resistance level of transgenic plants harboring the gene of interest from three vectors had no difference. The own promoter of Pi-d2, about 2.2 kb or 3.2 kb, had the similar promoter function as CaMV35S. Field evaluation for three successive years supported the results of artificial trial, and some lines with high resistance to rice leaf blast and neck blast were obtained.展开更多
LFY family genes play a conserved role in regulating flowering. In this study, the cDNA of LiLFY1 was isolated with the strategy of RT-PCR in combination with RACE from lily （Lilium longiflorum Thunb.）. LiLFY1 encod...LFY family genes play a conserved role in regulating flowering. In this study, the cDNA of LiLFY1 was isolated with the strategy of RT-PCR in combination with RACE from lily （Lilium longiflorum Thunb.）. LiLFY1 encodes a LFY transcriptional factor. The alignment analysis of the deduced LiLFY1 protein with other known LFY family proteins indicates that LiLFY1 is highly homologous with rice RFL and maize FLL. The result of Southern hybridization showed that there are two copies of LFY family genes in lily. LiLFY1 is expressed in young flower buds and shoot apical meristem （SAM） but not in roots, shoots, mature leaves, and mature floral organs. The cloned LiLFY1 gene may be applied to genetic engineering aiming for regulating the flowering time in lily.展开更多
基金supported by the Excellent Doctor Paper Foundation of the Ministry of Education of China (Grant No.200054)the Program for Innovative Research Team in University of China (Grant No.NCET-04-0907)the Program for New Century Excellent Talent in University of China (Grant No.IRT0453)
文摘The gene Pi-d2, conferring gene-for-gene resistance to the Chinese blast strain ZB15, was isolated from a rice variety （Digu） by the map-based cloning strategy. Here, we constructed a control plasmid pZH01-pi-d2tp309 （pZH01-tp309） and three different expression constructs, pCB-Pi-d25.3kb （pCB5.3kb）, pCB-Pi-d26.3kb （pCB6.3kb） and pZH01-Pi-d22.72kb （pZH01-2.72kb） of Pi-d2, driven by Pi-d2 gene’s own promoter or CaMV35S promoter. These constructs were separately introduced into japonica rice varieties Lijiangxintuanhegu, Taipei 309, Nipponbare and Zhonghua 9 through Agrobacterium- mediated transformation. A total of 150 transgenic rice plants were obtained from the regenerated calli selected on hygromycin. PCR, RT-PCR and Southern-blotting assay showed that the gene of interest had been integrated into rice genome and stably inherited. Thirty-five transgenic lines independently derived from T1 progeny were inoculated with the rice blast strain ZB15. Transformants exhibited resistance to rice blast at various levels. The lesions on the transgenic plant leaves were less severe than those on the controls and the resistance level of transgenic plants harboring the gene of interest from three vectors had no difference. The own promoter of Pi-d2, about 2.2 kb or 3.2 kb, had the similar promoter function as CaMV35S. Field evaluation for three successive years supported the results of artificial trial, and some lines with high resistance to rice leaf blast and neck blast were obtained.
基金the grant of National Center for Plant Gene Research,Institute of Genetics and Developmental Biology,Chinese Academy of Sciences,in Beijing
文摘LFY family genes play a conserved role in regulating flowering. In this study, the cDNA of LiLFY1 was isolated with the strategy of RT-PCR in combination with RACE from lily （Lilium longiflorum Thunb.）. LiLFY1 encodes a LFY transcriptional factor. The alignment analysis of the deduced LiLFY1 protein with other known LFY family proteins indicates that LiLFY1 is highly homologous with rice RFL and maize FLL. The result of Southern hybridization showed that there are two copies of LFY family genes in lily. LiLFY1 is expressed in young flower buds and shoot apical meristem （SAM） but not in roots, shoots, mature leaves, and mature floral organs. The cloned LiLFY1 gene may be applied to genetic engineering aiming for regulating the flowering time in lily.
