Establishment of a genetic transformation system for maize inbred P9-10

来源 :Chinese Science Bulletin | 被引量 : 0次 | 上传用户:lookingintheeye
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It was found that the supplement of 10~(-4) mol/L AgNO_3 in N6 medium enhanced the induction of type Ⅰembryogenic calli from immature embryos of maize inbred P9-10. After high-osmotic treatment, the induced calli was taken as transformation recipient to be bombarded with plasmid pMG6 carrying synthetic Bt gene by PDS-1000/He genegun. A total of 14 resistant calli were obtained after screening subsequently on the mediums with gradually increasing selective pressure. 10 plants were regenerated from the resistant calli on 3 different induction media,and 8 out of the 10 regenerated plants were confirmed to be integrated with Bt gene by PCR and Southern blotting analysis. Results of ELISA showed that the Bt-protein content in the leaves of the transgenic plants varied between 20—200 ng/g fresh weight. It was found that the supplement of 10 ~ (-4) mol / L AgNO_3 in N6 medium enhanced the induction of type I embryogenic calli from immature embryos of maize inbred P9-10. After high-osmotic treatment, the induced calli was taken as transformation recipient to be bombarded with plasmid pMG6 carrying synthetic Bt gene by PDS-1000 / He genegun. A total of 14 resistant calli were obtained after screening followed on the mediums with gradually increasing selective pressure. 10 plants were regenerated from the resistant calli on 3 different induction media, and 8 out of the 10 regenerated plants were confirmed to be integrated with Bt genes by PCR and Southern blotting analysis. Results of ELISA showed that the Bt-protein content in the leaves of the transgenic plants varied between 20-200 ng / g fresh weight.
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