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Kit comprising serial culture mediums and its application in soybean genetic transformation

A technology of genetic transformation and culture medium, applied to a kit composed of a series of culture medium and its application in soybean genetic transformation, can solve the problems of low transformation efficiency, restricting the development of soybean molecular breeding and the like

Active Publication Date: 2016-06-08
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing Agrobacterium tumefaciens transformation system has the problem of low transformation efficiency, which restricts the development of soybean molecular breeding

Method used

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  • Kit comprising serial culture mediums and its application in soybean genetic transformation
  • Kit comprising serial culture mediums and its application in soybean genetic transformation
  • Kit comprising serial culture mediums and its application in soybean genetic transformation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1, the preparation of kit

[0051] One, the preparation of kit provided by the invention

[0052] The kit provided by the invention consists of the following culture medium (each culture medium is individually packaged):

[0053] Germination medium (pH5.8): 3.12g / LB5 salt, 1ml / LB5 organic, 20g / L sucrose, 7.5g / L agar, the balance is water.

[0054] Liquid medium (pH5.4): 0.43g / LMS salt, 1ml / LB5 organic, 40mg / L acetosyringone, 150mg / L dithiothreitol, 100mg / LL-cysteine, 30g / L sucrose, 3.9mg / L 2-morpholineethanesulfonic acid, the balance is water.

[0055] Co-cultivation medium (pH5.4): 0.43g / LMS salt, 1ml / LB5 organic, 40mg / L acetosyringone, 150mg / L dithiothreitol, 100mg / LL-cysteine, 30g / L sucrose , 7.5g / L agar, 3.9mg / L 2-morpholineethanesulfonic acid, and the balance is water.

[0056] Recovery medium Ⅰ (pH5.4): 3.1g / LB5 salt, 1ml / LB5 organic, 30g / L sucrose, 150mg / L cephalosporin, 150mg / L Timentin, 1mg / L6-BA, 0.98g / L2 - Morpholinoethanesulfonic acid, 7.5g / L...

Embodiment 2

[0069] Embodiment 2, soybean genetic transformation

[0070] 1. Seed sterilization

[0071] 1. Take the plump, uniform and dry soybean variety Jack seeds without pests and spots, and spread them completely in a petri dish, and then put the petri dish into a desiccator.

[0072] 2. After completing step 1, put a 100ml beaker in the desiccator, pour 80ml of 12M sodium hypochlorite aqueous solution into the beaker, then slowly add 4ml of concentrated hydrochloric acid, then quickly cover the desiccator, seal it with Vaseline, and place it for 12 -16h, carry out chlorine gas sterilization.

[0073] 2. Preparation of infection solution

[0074] 1. Transform Agrobacterium tumefaciens EHA101 with pTF101.1 vector to obtain recombinant Agrobacterium.

[0075] 2. Take the recombinant Agrobacterium obtained in step 1 and resuspend it with the liquid medium prepared in Example 1 to obtain OD 600nm =0.6 of the infection bacteria solution.

[0076] Three, adopt the kit prepared in step...

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Abstract

The invention discloses a kit comprising serial culture mediums and its application in soybean genetic transformation. The kit comprises recovery culture medium, screening culture medium, growth culture medium, and rooting culture medium. The recovery culture medium comprises 45-55 mg / L of L- asparagine, 45-55 mg / L L-glutamine, 28-32 mg / L EDTA salt, and 11-13 mg / L FeSO4. The screening culture medium contains 45-44 mg / L L-asparagine, 45-55 mg / L L-glutamine, 28-32 mg / L EDTA salt, and 11-13 mg / L FeSO4. The growth culture medium contains 45-55 mg / L L-asparagine, 45-55 mg / L L-glutamine, 28-32 mg / L EDTA salt, and 11-13 mg / L FeSO4. The rooting culture medium contains 45-55 mg / L L-asparagine and 45-55 mg / L L-glutamine. The application of the kit in soybean genetic transformation belongs to the protection scope of this invention. The kit helps improve soybean genetic transformation efficiency and has high application value.

Description

technical field [0001] The invention relates to a kit consisting of a series of medium and its application in soybean genetic transformation. Background technique [0002] Soybean (Glycinemax L.Merr.) is an important grain and oil crop, and it is also the crop with the largest planting area of ​​transgenic varieties. Due to the limitations of traditional breeding methods, transgenic technology has become an important means of cultivating new soybean varieties. [0003] Currently, soybean transformation systems are widely used, mainly cotyledon node transformation system mediated by Agrobacterium tumefaciens and immature embryo transformation system mediated by particle gun. Since the transformation of immature embryos mediated by the particle gun is limited by the explant material, the cotyledonary node transformation method mediated by Agrobacterium tumefaciens is mainly used in practical applications. The existing Agrobacterium tumefaciens transformation system has the p...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H4/00A01H5/00
CPCA01H4/001A01H4/008C12N15/8205
Inventor 侯文胜陈莉韩天富姚伟伟陶金璐吴存祥孙石于洋郭艳翠
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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