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Birch efficient transgenic method

A transgenic, white birch technology, applied in horticultural methods, genetic engineering, plant genetic improvement, etc., can solve the problems of low transformation efficiency and complicated transformation process, and achieve the effect of accelerating the infection rate and enhancing the ability to resist Agrobacterium infection

Inactive Publication Date: 2013-06-12
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide a high-efficiency transgenic method for birch in order to solve the problems of complex transformation process and low conversion efficiency of the existing birch transgenic method

Method used

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  • Birch efficient transgenic method

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specific Embodiment approach 1

[0021] Specific embodiment one: a kind of white birch high-efficiency transgenic method of the present embodiment is carried out according to the following steps:

[0022] 1. Select rootless white birch seedlings with a height of 1-1.5 cm for subculture, culture them in the rooting medium for 24-26 days, remove the roots and move them into the rooting medium again for 30-35 days, and select seedlings with a height of 5-6 cm Leaf tissue at 0.2-0.3 cm from the base of the second or third leaf of the birch plant is used for Agrobacterium infection;

[0023] 2. Infect the leaves in step 1 by immersing them in the Agrobacterium bacterium liquid, wherein the bacterium liquid OD 600 0.1, the infection time is 3 to 5 minutes, place the infected leaves on sterile filter paper to absorb the attached bacterial solution, replace the filter paper and absorb the bacterial solution again;

[0024] 3. Inoculate the adaxial surface of the leaves dipped in step 2 on the callus induction medium...

specific Embodiment approach 2

[0028] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the rooting medium in step 1 includes WPM medium and IBA with a concentration of 0.4 mg / L. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0029] Embodiment 3: The difference between this embodiment and Embodiment 1 or 2 is that the leaf tissue used for Agrobacterium infection in step 1 is the leaf at 0.3 cm from the base of the second or third robust leaf. Others are the same as in the first or second embodiment.

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Abstract

The invention discloses a birch efficient transgenic method and relates to a plant transgenic method. The method disclosed by the invention can be used for solving the problems of low transformation efficiency and complex bacteria removing process of the existing birch transgenic method. The method is carried out through the following steps: step one, carrying out rooting culture twice for sub-cultured seedlings without root and selecting healthy leaves applied to agrobacterium infection; step two, infecting via agrobacterium; step three, co-culturing; and step four, finally acquiring transgenic plants through callus culture, differentiation culture, subculture and rooting culture. The method can be used for enhancing the agrobacterium infection-resisting capability of the leaves and speeds up the wound infection risk by selecting explants in the appropriate physiological state; the number of times for removing bacterium is reduced from the original 20-25 to 7-10; and the birch transgenic efficiency is increased by 2.32-3 times. The method can be applied to the birch transgenic field.

Description

technical field [0001] The invention relates to a method for plant transgene. Background technique [0002] Birch (Betula platyphylla Suk.) is one of the rare broad-leaved tree species in Northeast China. However, due to the long breeding cycle of birch and the slow progress of genetic improvement, traditional birch breeding techniques cannot design and transform traits according to breeding goals to meet the growing social needs. Therefore, it is imperative to carry out genetic engineering breeding research on birch. [0003] At present, the explant leaves selected by the existing white birch transgenic technology are too young or senile, and the young leaves are easily infected by Agrobacterium infection, and the explants need to be sterilized every day or every other day. It is not easy to obtain transformed cells; although senescent leaves are not easy to be infected by Agrobacterium, the callus differentiation ability of leaves is poor, and it is not easy to obtain tra...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H4/00A01H5/00
Inventor 姜静杨光韦睿刘桂丰
Owner NORTHEAST FORESTRY UNIVERSITY
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