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Quick and efficient transgenic method for white birch

A transgenic, birch technology, applied in horticultural methods, genetic engineering, plant genetic improvement, etc., can solve the problems of long transformation period and low efficiency of stable transformation of birch.

Inactive Publication Date: 2017-06-23
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims to solve the problems of low stable transformation efficiency and too long transformation period of existing white birch, and provides a platform for quickly studying the gene function of white birch and obtaining transgenic plants with improved target traits

Method used

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  • Quick and efficient transgenic method for white birch
  • Quick and efficient transgenic method for white birch
  • Quick and efficient transgenic method for white birch

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Experimental program
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specific Embodiment approach 1

[0019] Specific embodiment one: a kind of rapid and efficient white birch transgenic method of the present embodiment, it is carried out according to the following steps:

[0020] 1. Select white birch tissue culture seedlings with a seedling height of 5-6 cm, place the explant leaves and stem segments in the callus induction medium in an ultra-clean workbench, and induce culture for 7-10 days;

[0021] 2. Take out the callus grown in the callus induction medium in step 1, transfer it to the Agrobacterium tumefaciens infection solution containing the target gene sequence, vacuumize it with a vacuum pump for 5-10 minutes, and place it in a co-cultivation chamber. On the medium, dark treatment for 48h; among them, the infection solution OD 600 0.6~0.8,

[0022] 3. Place the callus after the dark treatment in step 2 on the transgenic resistance screening medium added with bacteriostatic agent, and carry out screening and culturing under light conditions at 25°C for 40 to 50 days...

specific Embodiment approach 2

[0024] Specific embodiment two: the difference between this embodiment and specific embodiment one is: the rooting medium described in step one contains the NAA (1-naphthylacetic acid) of WPM (Woody Plant medium), 0.2mg / L, 30g / L Sucrose and 3.0mg / L Phytagel (gellan gum), pH=5.8. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0025] Specific embodiment three: the difference between this embodiment and specific embodiment one is: the callus induction medium described in step one contains 6-BA (6-benzyladenine) of WPM (Woody Plant medium), 0.8mg / L, 0.5mg / L of GA 3 (Gibberellic acid A3), 0.02mg / L of NAA (1-naphthylacetic acid), 30g / L of sucrose and 3.0mg / L of Phytagel (gellan gum), pH=5.8. Others are the same as in the first embodiment.

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Abstract

The invention discloses a quick and efficient transgenic method for white birch, relates to a method for plant transgene, and aims at solving the problems that an existing method is low in stable transformation efficiency and long in transformation period of the white birch. The scheme provided by the invention is that the quick and efficient transgenic method comprises the following steps: selecting white birch tissue culture seedlings with the height of 5 to 6cm and carrying out induced culture on calluses; transferring the calluses into an agrobacterium tumefaciens infected solution containing a target gene sequence; carrying out vacuumized treatment and dark culture in sequence; screening resistant seedlings, and transferring the screened resistant seedlings into a rooting culture medium with a selective agent and a bacteriostatic agent added for culturing; finishing the operation after molecular detection is successfully verified. Compared with the prior art, the quick and efficient transgenic method has the advantages that the average transformation rate is improved by 2.71 times; in addition, the transformation period of infecting by using the calluses is shorted by one third or above compared with that of taking leaf blades and stems as explants. The quick and efficient transgenic method disclosed by the invention is applied to the field of transgenes.

Description

technical field [0001] The invention relates to a method for plant transgene. Background technique [0002] White birch (Betula platyphylla) is a deciduous tree of the genus Betula (Betula Linn). Widely distributed in eastern Asia. It is light-loving, cold-resistant, and has strong vitality. It is an excellent afforestation tree species for restoring vegetation and protecting forest water and soil. It is also an important economic tree species for medicinal and fiber use. However, the traditional breeding cycle of white birch is long and the process of genetic improvement is slow, which cannot meet the growing social needs by transforming traits according to the breeding objectives. In the genetic engineering breeding technology, the existing white birch transgene mainly transforms the explants directly, because the explants used in the regeneration and transformation of white birch are tissue-specific and the transformation period is long, which often leads to low transfo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84A01H4/00
CPCC12N15/8205
Inventor 刘中原高彩球王玉成张腾倩王培龙唐绯绯李亚博王媛媛
Owner NORTHEAST FORESTRY UNIVERSITY
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