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Tamarix hispida instantaneous transformation method

A technology of instant transformation of Tamarix tamarisk, applied in chemical instruments and methods, biochemical equipment and methods, botanical equipment and methods, etc., can solve the problems of long stable transformation period of T. tamarisk and low transformation efficiency of T. tamarisk transformation methods, and the like, To achieve the effect of short cycle

Inactive Publication Date: 2017-02-01
NORTHEAST FORESTRY UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The present invention aims to solve the problems that the existing stable transformation period of Tamarix bristles is too long and the transformation efficiency of the transformation method of Tamarix bristles is low, and provides a method for instantaneous transformation of Tamarix bristles

Method used

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  • Tamarix hispida instantaneous transformation method
  • Tamarix hispida instantaneous transformation method
  • Tamarix hispida instantaneous transformation method

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specific Embodiment approach 1

[0025] Specific embodiment one: the instant transformation method of Tamarix bristles in this embodiment, carries out according to the following steps:

[0026] 1. Select Tamarix bristles tissue-cultured seedlings subcultured for 30-35 days, with a plant height of 2.5-3.0 cm, and put the selected Tamarix bristles tissue-cultured seedlings into the high-osmotic treatment solution to soak for 5-10 minutes;

[0027] 2. Rapidly transfer the T. tumefaciens tissue culture seedlings treated with hypertonicity in step 1 to the infection solution containing the Agrobacterium tumefaciens containing the gene of interest, wherein the infection solution OD 600 0.6-0.8, put it in a vacuum pump to evacuate it for 5-10min, then place it in a constant temperature oscillator at 24-26°C, 100-130rpm for 4-5h;

[0028] 3. Take out the tissue culture seedlings after the infection in step 2, wash them with sterile water for 3-4 times, blot the water with sterile filter paper, insert the co-cultivati...

specific Embodiment approach 2

[0029] Embodiment 2: This embodiment differs from Embodiment 1 in that: the hyperosmotic treatment solution in Step 1 is 1.0 mol / L mannitol. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0030] Embodiment 3: This embodiment differs from Embodiment 1 or Embodiment 2 in that: the Agrobacterium tumefaciens containing the target gene in step 2 is transformed with the Agrobacterium tumefaciens containing the expression vector of the target gene. Others are the same as in the first or second embodiment.

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Abstract

The invention discloses a tamarix hispida instantaneous transformation method, and relates to a plant instantaneous transgenosis method. The invention aims at solving the problems that an existing tamarix hispida stable transformation cycle is too long and an original tamarix hispida instantaneous transformation method is low in transformation efficiency. The method comprises the following steps: I, selecting tamarix hispida tissue culture seedlings obtained from subculture, and soaking the selected tamarix hispida tissue culture seedlings in high-permeability treatment liquid; II, rapidly transferring the tamarix hispida tissue culture seedlings which have undergone high-permeability treatment into agrobacterium tumefaciens infection liquid which contains a target gene, conducting vacuumizing in a vacuum pump and conducting infection in a constant-temperature oscillator; and III, taking out the infected tissue culture seedlings, washing the tissue culture seedlings, sucking water, inserting the tissue culture seedlings into a sodium co-culture medium and conducting co-culture. With the application of the method, the instantaneous transformation efficiency of the tamarix hispida is greatly improved. On the basis of an enzyme activity determination result of a marker gene GUS instantaneously transformed gene, it shows that the method, in comparison with the original tamarix hispida instantaneous transformation method, can improve a GUS activity by 2.45 times. The method provided by the invention is applicable to the field of plant instantaneous transgenosis.

Description

technical field [0001] The invention relates to a method for instantaneous transgenic plants. Background technique [0002] Tamarix hispida is a shrub or small tree in the genus Tamarix of the family Tamarix. It has strong drought resistance, salt tolerance, water and humidity resistance, and sand burial resistance. At the same time, due to its excellent resistance to stress, it is an ideal material for studying the mechanism of plant stress resistance and cloning of stress resistance genes. However, due to the long genetic transformation cycle of Tamarix, it is difficult and time-consuming to study the stress resistance function and mechanism of the gene by overexpressing and RNAi inhibiting the expression of the target Tamarix gene. Finding a fast and effective genetic transformation method is of great significance for the study of gene function in Tamarix. Contents of the invention [0003] The invention aims to solve the problems that the existing stable transformati...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H5/00
CPCC07K14/415C12N15/8205C12N15/8271
Inventor 高彩球王玉成刘中原赵玉琳及晓宇王培龙
Owner NORTHEAST FORESTRY UNIVERSITY
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