Gentic transforming method of perennial rye grass
A technology of ryegrass and purpose, applied in the field of genetic transformation of perennial ryegrass, can solve the problems of few reports on genetic transformation of ryegrass, large variation of cloned regenerated plants, difficulty in establishing protoplasts and protoplast regeneration systems, etc.
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[0028] 1. Materials
[0029]Perennial ryegrass tested materials mainly include Accent, Derby, Supreme Derby, Gold Medalist, Top Gun, Duofu, Emerald (Green Gem), purchased from the market.
[0030] Tobacco varieties, mature seeds of Honghua Dajinyuan, can be purchased from the market. Used to increase conversion efficiency.
[0031] Agrobacterium tumefaciens strain EHA105, resistant to the antibiotic rifampicin (Rif), carries the binary vector plasmid pCAMBIA-2301, the expression cassette of which is shown in figure 1 . Such as figure 1 As shown, the plasmid carries the selection gene neomycin phosphotransferase gene (npt II) and the reporter gene β-glucuronidase gene (gusA), and gusA contains two catalase introns, so that it cannot be expressed in Agrobacterium. These two genes are both activated by the cauliflower mosaic virus 35S promoter, can replicate and express in plant cells, and can be directly introduced into Agrobacterium to transform plants. This plasmid is m...
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Application Information
- IPC
- A01H1/00
- Inventors
- 马欣荣; 刘华玲