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Agrobacterium tumefaciens mediated genetic transformation method for maize backbone inbred line

A technology of genetic transformation and inbred lines, applied in the field of biology

Active Publication Date: 2020-05-15
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But for the corn backbone inbred line Jing 724, there is no report on its efficient tissue culture and genetic transformation methods

Method used

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  • Agrobacterium tumefaciens mediated genetic transformation method for maize backbone inbred line
  • Agrobacterium tumefaciens mediated genetic transformation method for maize backbone inbred line
  • Agrobacterium tumefaciens mediated genetic transformation method for maize backbone inbred line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1, the medium involved in the tissue culture and genetic transformation of the corn backbone inbred line Jing 724

[0086] 1. Callus induction medium

[0087] The medium designed for the callus induction of the corn backbone inbred line Jing 724 includes callus induction medium MC-1, callus induction medium MC-2 and callus induction medium MC-3.

[0088] The callus induction medium MC-1 is composed of solute and solvent, the solvent is double distilled water, the solute and its concentration are: 0.5ml / L 1000X N6 basic salt, 2g / L MS basic salt, 1ml / L 1000X N6 organic, 0.2g / L Calcium Chloride Dihydrate, 20g / L Sucrose, 1.4g / L Proline, 0.5g / L Hydrolyzed Casein, 0.1g / L Inositol, 0.5mg / L 2,4-D, 2.5mg / L dicamba, 0.2mg / L 6-benzylaminopurine, 3g / L plant gel. The pH value of callus induction medium MC-1 was 5.8.

[0089] Callus induction medium MC-2 is composed of solute and solvent, the solvent is double distilled water, the solute and its concentration are: 0.35ml / ...

Embodiment 2

[0107] Example 2, Agrobacterium-mediated genetic transformation method of maize backbone inbred line Jing 724 immature embryo

[0108] 1. Carrier construction

[0109] Insert the pr2X35s::GFP::t35s / / prZmUbi-01::HPT::tNos fragment shown at position 131-5144 of Sequence 1 into the T-DNA region of the transformed pCAMBIA3301 vector to obtain the pY105 recombinant vector. The nucleotide sequence of the pY105 recombinant vector is shown as sequence 1 in the sequence listing.

[0110] 2. Preparation of Agrobacterium

[0111] The pY105 vector prepared in step 1 was introduced into EHA105 (EHA105 is a product of Shanghai Weidi Biotechnology Co., Ltd., CAT#:AC1010) Agrobacterium, and after identification, EHA105 Agrobacterium containing the pY105 vector was obtained. The EHA105 Agrobacterium liquid containing the pY105 vector was streak-coated on the Agrobacterium activation medium (the medium in Step 2 of Example 1), and cultivated in the dark at 28° C. for 36-48 hours.

[0112] 3....

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Abstract

The invention discloses an Agrobacterium tumefaciens mediated genetic transformation method for a maize backbone inbred line Beijing 724. The method comprises the following steps: 1) infecting embryosof maize inbred line Beijing 724 with vector Agrobacterium tumefaciens containing a target gene, so as to obtain infected embryos; 2) inoculating a coculture medium with the infected embryos for culture, so as to obtain co-cultured embryos; 3) inoculating a recovery culture medium with the co-cultured embryos for culture, so as to obtain recovery-cultured embryos; 4) inoculating a screening culture medium with the recovery-cultured embryos for culture, so as to obtain resistant calli; 5) inoculating a regeneration culture medium with the resistant calli for culture, so as to obtain young seedlings; and 6) culturing the young seedlings in a rooting culture medium, thereby obtaining transgenic plants of the maize inbred line Beijing 724.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for genetic transformation of a corn backbone inbred line, in particular to a method for genetic transformation of a corn backbone inbred line Jing 724 mediated by Agrobacterium. Background technique [0002] Corn is an important crop for the supply of food, feed, and industrial raw materials. It plays an important role in the modern era of population growth, animal husbandry, and high-speed industrial development. According to the statistics of the Ministry of Agriculture, the gap between my country's corn supply and demand in 2017 / 2018 was 4.3 million tons. According to the analysis of the corn industry development trend report, the growth rate of China's corn production will decline during 2016-2030, while the growth rate of total corn consumption It will obviously exceed the growth rate of production, and the gap between domestic supply and demand will further...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00A01H6/46A01H4/00
CPCC12N15/8205A01H4/008
Inventor 武莹贺晓庆宋金岭杨进孝
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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