Transformation method of dicotyledonous plant cells
A dicotyledonous plant and cell technology, applied in the field of plant biology, can solve problems such as indistinguishable diploid events and polyploid event sterility
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example 1
[0064] Example 1: Transformation method in tomato
[0065] The following are example methods for transforming dicots. This method uses tomatoes as an example. The method was changed in some instances, as shown in the related instance.
[0066] (1) Disinfection of seeds
[0067] a. Dried mature tomato seeds at least 14 days post-pollination were soaked in 10% Clorox with 0.04% silwet-77 for 20 min. Then rinse the seeds with sterile water at least 3 times.
[0068] (2) Seed infiltration
[0069] a. Transfer the sterilized tomato seeds to germination medium (Gamborg B5 medium containing 20 g / L sucrose, 8 g / L agar, pH 5.6) or sterile filter paper soaked in sterile water. The seeds were then incubated in the dark at 25°C for 1 to 3 days (before the cotyledons emerged from the seed coat). One day of incubation is preferred.
[0070] (3) Preparation for vaccination
[0071]Agrobacterium strain EHA101 containing the vector of interest stored at -80°C was streaked with appropri...
example 2
[0097] Example 2: Transformation of explants from tomato seedlings soaked for 1 day and seedlings soaked for 7 days
[0098] Variety used: Moneymaker
[0099] Carrier used: 24416
[0100] The transformation procedure for explants from seeds at least 14 days post-pollination, 1 day infiltration was as described in Example 1 .
[0101] Procedure for transformation of explants from 7-day seedlings:
[0102] 1. Sterilization of seeds: as described in Example 1.
[0103] 2. Seed infiltration: Transfer the sterilized seeds to a growth vessel containing an appropriate amount of germination medium (0.5X MS medium, containing 10 g / L sucrose, 10 g / L Phytagel, pH 5.8). . Seeds were kept at 25°C for 16h / 8h (light / dark) for 7 days.
[0104] 3. Agrobacterium: An Agrobacterium suspension was prepared as described in Example 1 .
[0105] 4. Explant preparation and infection: Hypocotyls and cotyledons were cut into segments approximately 5 mm long. The segments were inoculated in the Ag...
example 3
[0119] Example 3: Transformation of Different Explant Types
[0120] Variety used: Syngenta Variety 1
[0121] Carrier used: 24133
[0122] Source of explants: Tomato seeds at least 14 days after pollination and 1 day of infiltration
[0123] The transformation procedure was performed as described in Example 1.
[0124] Table 4. Results
[0125]
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