Method for establishing agrobacterium rhizogenes-mediated vegetable pea genetic transformation system
A genetic transformation system, the technology of Agrobacterium rhizogenes, is applied in the field of plant genetic engineering to achieve the effects of saving experimental costs, efficient and faster experimental operations, and reducing pollution rates
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[0029] 1) preparation of the explant of the above-ground part of vegetable pea;
[0030] Select mature vegetable pea seeds that are free from diseases and insect pests, disinfect them with 75% sodium hypochlorite for 15 minutes, rinse them with sterile water for 5-6 times, and sow them in the substrate (the preparation of the substrate: mix vermiculite and peat in a ratio of 3 to 1, and heat them at 121° C. , autoclaved for 20 min, poured with sterile water), placed in a light incubator (22°C, photoperiod 16h light / 8h dark) for cultivation. After growing for about one week (5 days, 7 days, 9 days), the plants were cut off at the upper and lower ends of the cotyledon adjacent to the plants respectively, to obtain the explants of the aboveground part of the vegetable pea.
[0031] 2) Activation and cultivation of AR1193 Agrobacterium rhizogenes strain;
[0032] ① Take 200 μL of Agrobacterium rhizogenes strain AR1193 glycerol bacterium (carrying the binary expression vector pCAM...
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