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
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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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