科研之友
微信
新浪微博
Facebook
分享链接
摘要
A reliable Agrobacterium-mediated transformation and shoot regeneration protocol was developed for breeding lines of commercially important melon. Genetic manipulation has been considered a feasible approach for melon improvement; however, melon is considered a crop species difficult to manipulate. Here we proposed meristematic cells from mature embryos as target for gene transfer by Agrobacterium. In vitro meristems proliferation and multiple shoots regeneration were evaluated by sowing melon mature seeds on MS with 1.0 mg/L benzyladenine (BA), and 0.05 mg/L indole acetic acid (IAA) were used for shoot regeneration. The highest number of regenerated shoots was obtained from half mature seeds. A DNA fragment corresponding to selection marker nptII was amplified from genomic DNA extracted from leaves of regenerated plant on hormone free MS medium with 75 mg/L kanamycin, suggesting their transgenic nature. Southern hybridization of transgenic lines revealed random insertion of the transgene in host genome, with insert numbers differing among transformants anthesis, suggesting that ethylene is important for sex determination. Field studies showed that CmACS-7 melons had earlier mature bisexual flowers, increased femaleness as measured by earlier and bisexual buds, and increased number of fruit set on closely spaced nodes on the main stem. Transformation efficiencies of cultivar CM-23 with EHA105 (pBI121-cm) were 4 %, demonstrating that melon meristematic cells are an useful target for genetic manipulation by agroinfection.
- 出版日期2014-7
- 单位东北农业大学
