Efficient genetic transformation method of whole strain infection mediated by Agrobacterium tumefaciens in Rhododendron alpinum L

A genetic transformation method, Agrobacterium-mediated technology, applied in biochemical equipment and methods, botanical equipment and methods, introduction of foreign genetic material using vectors, etc. However, the research started late, and the total amount is dependent on foreign imports, achieving the effects of great convenience and reliability, shortening the breeding process, and shortening the transformation period.

Active Publication Date: 2019-01-15
FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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  • Abstract
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Problems solved by technology

There are more than 320 species of rhododendrons in my country, accounting for 1/3 of the world's rhododendrons. The southwest and adjacent eastern Himalayas are the distribution centers and origins of the evergreen rhododendron subgenus, but the breeding research on rhododendrons started late. , lagging far behind the western countries, the Rhododendron species and their total quantity sold on the market in our country are still in the embarrassing situation of relying on foreign imports to a large extent
In this situation, in addition to conducting conventional cross-breeding research on Rhododendron plants, researchers must spee

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  • Efficient genetic transformation method of whole strain infection mediated by Agrobacterium tumefaciens in Rhododendron alpinum L
  • Efficient genetic transformation method of whole strain infection mediated by Agrobacterium tumefaciens in Rhododendron alpinum L
  • Efficient genetic transformation method of whole strain infection mediated by Agrobacterium tumefaciens in Rhododendron alpinum L

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Embodiment 1

[0044] In this embodiment, Rhododendron delavayi in alpine rhododendron is taken as an example, and the efficient genetic transformation method of the whole plant infection method mediated by Agrobacterium comprises the following steps:

[0045] (1) Seed sterilization and seed pre-cultivation

[0046] ① Seed sterilization, the seeds of Rhododendron lantana with ddH 2 O wash 2 times, then soak the seeds with 75% v / v alcohol for 45 seconds, and then wash with ddHO 2 Wash twice with O; then soak Rhododendron alpine seeds in 2% v / v sodium hypochlorite solution with Tween-20 for 15 minutes, shake well 2 to 3 times during the period, and finally rinse with ddH 2 O cleaning seeds 2 to 3 times; the 2% v / v sodium hypochlorite solution added with Tween-20 is to add 2 to 3 drops of Tween-20 for every 10 ml of 2% v / v sodium hypochlorite solution.

[0047] ②Seed pre-cultivation

[0048] Soak the sterilized Rhododendron lantana seeds in pH=5.2 containing 0.03g / L GA 3In the 1 / 2MS liquid ...

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Abstract

The invention discloses an efficient genetic transformation method of whole strain infection method mediated by Agrobacterium tumefaciens of Rhododendron alpinum. The method mainly comprises the following steps: seed pre-culture, ultrasonic treatment, preparation of Agrobacterium tumefaciens bacterial solution, agrobacterium infection and vacuum filtration, co-culture, degerming washing, germination culture, GUS histochemical staining. The method of the invention has the advantages of simple operation, low experiment cost, short transformation period and high genetic transformation efficiencyof 61%, and can be applied to the genetic transformation of other plants of Rhododendron.

Description

technical field [0001] The invention relates to the technical field of plant life sciences, in particular to a high-efficiency genetic transformation system invented by using Rhododendron delavayi, an alpine rhododendron, as an experimental material. Background technique [0002] Azalea (Rhododendron L.) is a world-renowned garden ornamental plant, one of the top ten traditional flowers in my country and one of the eight famous flowers in Yunnan Province, China. Alpine Rhododendron is a general term for some original species of evergreen broad-leaved Rhododendron and its hybrid offspring. Because of its beautiful plant shape, large and colorful flowers, and evergreen seasons, it has gradually become a rookie in my country's New Year's Eve flowers and high-end landscaping. There are more than 320 species of rhododendrons in my country, accounting for 1 / 3 of the world's rhododendrons. The southwest and adjacent eastern Himalayas are the distribution centers and origins of the ...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00A01H6/36
CPCC12N15/8205
Inventor 张露王继华宋杰李绅崇蔡艳飞李世峰
Owner FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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