High-frequency plant regeneration method of rice cropping mature embryo

A technology for mature embryos and rice, applied in the field of Agrobacterium-mediated gene transformation, which can solve the problems of incompatibility of improvement and inappropriate use of large-scale test-tube seedlings for rice genetic engineering screening, so as to reduce the time for development and shorten the breeding cycle , cost reduction effect

Inactive Publication Date: 2009-09-23
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is: aiming at the high dependence on the genotype in the regeneration technology of cultivated rice mature embryos, a kind of rice material has only one specific optimum medium formula, and the development of each medium formula and culture technology requires Half a year, or even longer, is not suitable for the improvement of a large number of existing rice cultivars, and is not suitable for large-scale test-tube seedling propagation and future rice genetic engineering screening and other defects.

Method used

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  • High-frequency plant regeneration method of rice cropping mature embryo
  • High-frequency plant regeneration method of rice cropping mature embryo
  • High-frequency plant regeneration method of rice cropping mature embryo

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Selection of callus induction medium

[0029] Selection of callus induction medium:

[0030] M8 Major elements, trace elements and organic components, iron salts are the basic components, and 2mg / L of 2,4-dichlorophenoxyacetic acid is added;

[0031] N6 macroelements, trace elements and organic components, iron salts are the basic components, and 2mg / L of 2,4-dichlorophenoxyacetic acid is added;

[0032] MS macroelements, trace elements and organic components, iron salts are the basic components, and 2mg / L of 2,4-dichlorophenoxyacetic acid is added;

[0033] Rice: The seeds of Wuyujing 3 harvested in the same year are used as explant materials.

[0034] experiment method:

[0035] The mature seeds of Wuyujing No. 3 were dehulled, treated with 75% ethanol for 5 minutes, soaked in 0.1% mercuric chloride solution for 15 minutes, rinsed with sterile water for 4-5 times, and cultured overnight at 26°C±2°C. The next day, it was treated with 75% ethanol for 5 min...

Embodiment 2

[0038] The substitution effect of commercially available white granulated sugar to different single carbon sources of embodiment 2

[0039] carbon source choice

[0040] M8 Macroelements, trace elements and B 5 Vitamins are the basic components, add mass ratio 8% agar strips, 0.5% sorbitol, 2mg / L 2,4-dichlorophenoxyacetic acid, 0.5mg / L 6-benzyl adenine, select mass ratio 3% commercially available white granulated sugar is used as carbon source (white granulated sugar is purchased from Jiangsu Nanjing Suguo Supermarket, 3.5 yuan / 500g);

[0041] M8 Macroelements, trace elements and B 5 Vitamins are the basic components, add mass ratio 8% agar strips, 0.5% sorbitol, 2mg / L 2,4-dichlorophenoxyacetic acid, 0.5mg / L 6-benzyl adenine, select mass ratio 3% sucrose as carbon source (purchased from Shanghai Kaimi Chemical Co., Ltd., price: 82 yuan / 500g);

[0042] M8 Macroelements, trace elements and B 5 Vitamins are the basic components, add mass ratio 8% agar strips, 0.5% sorbitol, ...

Embodiment 3

[0050] Example 3 Induction of Callus from Different Rice Mature Embryos

[0051] Test group callus induction medium:

[0052] Indica rice callus induction medium: based on the basic components of macroelements, organic components and trace elements of M8, add 3% white sugar, 0.8% agar strips, 0.5% sorbitol, 1mg / L 2,4-dichlorophenoxyacetic acid, 1mg / L abscisic acid, 1.5mg / L 6-benzyladenine and 0.2mg / L kinetin;

[0053] Japonica rice callus induction medium: based on the basic components of macroelements, organic components and trace elements of M8, add 3% white sugar, 0.8% agar strips, 0.5% sorbitol, 2mg / L 2,4-dichlorophenoxyacetic acid, 1mg / L abscisic acid, 0.5mg / L 6-benzyladenine and 0.2mg / L kinetin;

[0054] Hybrid rice callus induction medium: based on the basic components of macroelements, organic components and trace elements of M8, add 3% white sugar, 0.8% agar strips, 0.5% sorbitol, 2mg / L 2,4-dichlorophenoxyacetic acid, 1mg / L abscisic acid, 1mg / L 6-benzyl adenine and...

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Abstract

The invention relates to a high-frequency plant regeneration method of rice cropping mature embryo, which is characterized by comprising the following steps: adopting white granulated sugar as compound carbon sources in callus induction culture medium, successive basic culture medium, differential medium and seedling strong root culture medium of rice; performing all-black culture 7 days before callus induction culture, and then switching to culture under light, obtaining callus; selecting pale yellow, dried and granular callus to be switched to the successive basic culture medium for culturing under light; selecting fluey callus to be switched to the differential medium for culturing under culturing light; selecting green shoot containing granular callus to be switched to the seedling strong root culture medium for continuously culturing, and forming strong tube plant.

Description

technical field [0001] The invention relates to a technical method for cultivating mature rice embryos for high-frequency regeneration plants, which belongs to the field of biotechnology and is generally applicable to the tissue culture of mature rice embryos cultivated by different genotypes such as indica rice, japonica rice and hybrid rice, screening of somatic cell mutants and Agrobacterium-mediated gene transformation. Background technique [0002] In recent years, due to the increase of population and the reduction of arable land, the demand for increasing the yield of rice, which is more than half of the world's staple food, has become more urgent, but in fact, due to the wide application of dwarf genes and heterosis utilization in my country, rice breeding Great achievements have been made, and the space for improving rice through conventional breeding is getting smaller and smaller. In the future, there is an urgent need to adopt more efficient technologies and new ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 李霞阎丽娜周月兰
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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