Agrobacterium‐mediated transformation of Japanese lawngrass (Zoysia japonica Steud.) containing a synthetic cryIA(b) gene from Bacillus thuringiensis
作者： Lei Zhang , D. Wu , Li Zhang , C. Yang
DOI: 10.1111/J.1439-0523.2007.01320.X
关键词:
摘要: Transgenic Japanese lawngrass plants containing a synthetic cryIA(b) gene from Bacillus thuringiensis under the control of maize ubiquitin promoter were developed by Agrobacterium-mediated transformation. A total 1540 embryogenic calluses derived dormancy-removal mature seeds co-cultured with disarmed strain EHA105 harhouring binary vector pKUB. Three days after co-culture in dark at 21°C, transient β-glucuronidase (GUS) expression frequency was 74.2%. After selection 100 mg/l hygromycin B, over 50 independent resistant cell clones and 25 regenerated obtained. The integration into genome confirmed 22 GUS histochemical assay, PCR amplification, Southern blotting Western analysis, transformation efficiency 1.4%. entire process callus induction to production transgenic plantlets 80-100 days. T 1 progeny segregation analysis these lines demonstrated that 59.1% events inherited typical Mendelian fashion.
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