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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

Pending Publication Date: 2022-07-08
SYNGENTA PARTICIATIONS AG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Polyploid events are usually sterile and phenotypically normal, indistinguishable from diploid events at an early stage

Method used

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  • Transformation method of dicotyledonous plant cells
  • Transformation method of dicotyledonous plant cells
  • Transformation method of dicotyledonous plant cells

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The present disclosure relates to methods of transforming dicotyledonous plants such as Solanum lycopersicum and Watermelon. In some aspects, the methods include using dicotyledonous explants obtained by segmenting the seed and soaking the dicotyledonous explants in various media. In some aspects, transformed cells of dicotyledonous plants and dicotyledonous plants comprising transformed cells are provided.

Description

[0001] Related applications [0002] This application claims priority to provisional application PCT / CN2019 / 104384, filed on September 4, 2019 and incorporated herein by reference in its entirety. technical field [0003] The present invention generally relates to the field of plant biotechnology. More specifically, the present invention relates to methods for improving transformation in dicots, eg, improving transformation frequency and / or reducing polyploidy events in dicots. Background technique [0004] Tomato (Solanum lycopersicum L.) is the second most important vegetable crop after potato. Eating tomatoes is considered to have a significant contribution to human health and nutrition because tomatoes are rich in minerals, vitamins, and lycopene and beta-carotene pigments. [0005] The transformation system of tomato was established in the 1980s, and various transformation systems have been reported successively since then. In most transformation systems, cotyledons ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H5/08
CPCC12N15/8205A01H5/02C12N15/8207
Inventor 许建平李超文琴
Owner SYNGENTA PARTICIATIONS AG