High embryogenic calli of three cutivars of Kentucky bluegrass, Md, Bd, and Gm, were induced from mature embryos, and were proliferated on medium K3 and K5. Embryogenic calli were transformed with plamids pDM803 and p...High embryogenic calli of three cutivars of Kentucky bluegrass, Md, Bd, and Gm, were induced from mature embryos, and were proliferated on medium K3 and K5. Embryogenic calli were transformed with plamids pDM803 and pBY520 by microprojectile bombardment. Fourty-two transgenic lines had been obtained. The highest efficiency of transformation reached to 3.7% for cv. Md, 2.8% for cv. Gm, and 5 % for cv. Bd. The micro nutriment of Cupric had significant effect on transformation. The embryogenic callus cultured in dim-light condition had higher transformation efficiency than the green callus cultured in light condition for one month before transformation. The selective regime and selective pressure on the putative transgenic plants were important for obtaining the desire number of transgenic plants. It also affected the copy number of integrated genes in the genomic DNA of transgenic plants.展开更多
A protocol was discussed for high efficient plant regeneration from seven bluegrass （Poa pratensis L.） cultivars via an in- direct callus induction and somatic embryogenesis method. Mature seeds were used as explant...A protocol was discussed for high efficient plant regeneration from seven bluegrass （Poa pratensis L.） cultivars via an in- direct callus induction and somatic embryogenesis method. Mature seeds were used as explants for callus initiation. Callus induction and proliferation efficiencies were investigated on NB, modified MS （MMS） and MS media, supplemented with 2.0 mg·L^-1 2,4-dichlorophenoxyacetic acid （2,4-D）. The MMS medium performed best. Based on the MMS medium, direct and indirect callus induction effects of bluegrass from mature seeds were compared at the range of 1-5 mg·L^-1 2,4-D contained in the medium. Under the direct callus induction method, the most suitable 2,4-D concentrations varied among cultivars. Under the indirect callus induction method, a significantly high callus induction frequency （93.33%-98.33%） was obtained and there were barely any statistically sig- nificant differences among the tested genetically diverse cultivars. Somatic embryos were promoted on the MMS medium supple- mented with 3 mg·L^-1 2,4-D, 0.1 mg·L^-1 kinetin and 0.8 mg·L^-1 CuSO4. Embryogenetic calli developed into plantlets on the MMS medium containing different concentrations of thidiazuron （TDZ）, and the differentiation frequencies varied in the range from 20.15% to 77.65%. The 0.25 mg·L^-1 TDZ was generally the most suitable concentration for the tested cultivars.展开更多
基金This project is supported by the Foundation of State Key Laboratory of Freshwater Ecology and Biotechnology,China(2001FB10)Young Foundation of Shanxi,China(20001038)+2 种基金Shanxi Scholarship Foundation,Chinathe Hational Natural Science Foundation of China(30270204)DLF-Trifolium,Research Division,Denmark.
文摘High embryogenic calli of three cutivars of Kentucky bluegrass, Md, Bd, and Gm, were induced from mature embryos, and were proliferated on medium K3 and K5. Embryogenic calli were transformed with plamids pDM803 and pBY520 by microprojectile bombardment. Fourty-two transgenic lines had been obtained. The highest efficiency of transformation reached to 3.7% for cv. Md, 2.8% for cv. Gm, and 5 % for cv. Bd. The micro nutriment of Cupric had significant effect on transformation. The embryogenic callus cultured in dim-light condition had higher transformation efficiency than the green callus cultured in light condition for one month before transformation. The selective regime and selective pressure on the putative transgenic plants were important for obtaining the desire number of transgenic plants. It also affected the copy number of integrated genes in the genomic DNA of transgenic plants.
文摘A protocol was discussed for high efficient plant regeneration from seven bluegrass （Poa pratensis L.） cultivars via an in- direct callus induction and somatic embryogenesis method. Mature seeds were used as explants for callus initiation. Callus induction and proliferation efficiencies were investigated on NB, modified MS （MMS） and MS media, supplemented with 2.0 mg·L^-1 2,4-dichlorophenoxyacetic acid （2,4-D）. The MMS medium performed best. Based on the MMS medium, direct and indirect callus induction effects of bluegrass from mature seeds were compared at the range of 1-5 mg·L^-1 2,4-D contained in the medium. Under the direct callus induction method, the most suitable 2,4-D concentrations varied among cultivars. Under the indirect callus induction method, a significantly high callus induction frequency （93.33%-98.33%） was obtained and there were barely any statistically sig- nificant differences among the tested genetically diverse cultivars. Somatic embryos were promoted on the MMS medium supple- mented with 3 mg·L^-1 2,4-D, 0.1 mg·L^-1 kinetin and 0.8 mg·L^-1 CuSO4. Embryogenetic calli developed into plantlets on the MMS medium containing different concentrations of thidiazuron （TDZ）, and the differentiation frequencies varied in the range from 20.15% to 77.65%. The 0.25 mg·L^-1 TDZ was generally the most suitable concentration for the tested cultivars.
