Culture medium for overcoming variety and genetype restriction in regeneration culture of alfalfa high frequency somatic embryos

A technology for genotype of varieties and regeneration medium, applied in the field of botany, can solve the problems of great difference in genotype, low self-seeding rate, difficulty in solving alfalfa genotype obstacles, etc., achieve efficient and stable genetic transformation, and improve breeding efficiency. , the effect of increasing the frequency of regenerated plants

Inactive Publication Date: 2010-05-12
JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY +1
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Problems solved by technology

[0003] Alfalfa is a perennial cross-pollinated plant with a very low self-seed setting rate, less than 1%. Most alfalfa varieties currently used in cultivation are comprehensive varieties, and the genotypes of individuals within a variety vary greatly. It can be said that each A single seed is a single genotype, and the prerequisite for cultivar improvement using biotechnology is the ability to regenerate high-frequency somatic embryos in the tissue culture process, but the somatic embryogenesis ability of alfalfa is strongly dependent on its genotype, so the genotype Obstacles have become bottlenecks that limit the high-frequency somatic embryo regeneration of alfalfa, especially high-quality alfalfa
[0004] There have be

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  • Culture medium for overcoming variety and genetype restriction in regeneration culture of alfalfa high frequency somatic embryos
  • Culture medium for overcoming variety and genetype restriction in regeneration culture of alfalfa high frequency somatic embryos
  • Culture medium for overcoming variety and genetype restriction in regeneration culture of alfalfa high frequency somatic embryos

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specific Embodiment approach

[0067] figure 1 It is a schematic diagram of Jinda embryogenic callus according to the present invention; figure 2 It is a schematic diagram of Suntory granular embryogenic callus according to the present invention; image 3 It is a schematic diagram of the development of Jinda somatic embryos into globular embryos, pear-shaped embryos and torpedo embryos according to the present invention; Figure 4 It is a schematic diagram of the development of Zhongmu No. 1 somatic embryos into spherical embryos, pear-shaped embryos and torpedo embryos according to the present invention; Figure 5 It is a schematic diagram of embryoid bodies in different stages of Zhongmu No. 1 according to the present invention; Figure 6 It is a schematic diagram of the differentiation and aggregation of Jinda embryoid bodies into bud clusters according to the present invention; Figure 7 It is a schematic diagram of the differentiation and aggregation of Suntory embryoid bodies into bud clusters acc...

Embodiment 1

[0114] A. Embryogenesis callus induction medium (improved SH+6mg / L 2,4-D+0.2mg / L 6-BA+30g / L sucrose+0.3%phytagel)+B. Embryo subculture formation medium ( Improved SH+5mg / L 2,4-D+0.2mg / L 6-BA+500mg / L hydrolyzed casein+50g / L sucrose+0.35% phytagel)+C. Embryo maturation and germination medium (improved MS+ 500mg / L hydrolyzed casein + 30g / L sugar + 0.7% agar) + D. Seedling growth acclimatization medium (1 / 2 improved MS + 20g / L sugar + 0.7% agar), the pH of all the above-mentioned media after autoclaving Around 5.8.

Embodiment 2

[0116] A. Somatic embryogenesis induction medium (improved SH+6mg / L 2,4-D+0.5mg / L 6-BA+30g / L sucrose+0.3%phytagel)+B. Embryo subculture formation medium (improved SH+5mg / L 2,4-D+0.5mg / L 6-BA+500mg / L hydrolyzed casein+50g / L sucrose+0.35% phytagel)+C. Embryo maturation and germination medium (improved MS+500mg / L hydrolyzed casein+30g / L white sugar+0.7% agar)+D. Seedling growth and acclimatization medium (1 / 2 improved MS+20g / L white sugar+0.7% agar), the pH of all above-mentioned mediums after autoclaving is at Around 5.8.

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Abstract

The invention discloses a culture medium for overcoming variety and genetype restriction in regeneration culture of alfalfa high frequency somatic embryos. The culture medium comprises seedling growth and acclimatization culture medium, a somatic cell embryo occurrence and induction culture medium, an embryo subgeneration formation culture medium and an embryo maturing and sprouting culture medium. The seedling growth and acclimatization culture medium is half modified MS+20g/L white sugar+0.7% agaragar; the somatic cell embryo occurrence and induction culture medium is modified SH+6-10mg/L2,4-D+0.2-0.5mg/L 6-BA+30g/L cane sugar+0.3% phytagel; the embryo subgeneration formation culture medium is modified SH+2-5 mg/L 2,4-D+0.2-0.5mg/L6-BA+500mg/L hydrolyzed casein+50g/L cane sugar+0.35% phytagel; and the embryo maturing and sprouting culture medium is modified MS+500mg/L hydrolyzed casein+30g/L white sugar+0.7% agaragar. The invention can effectively overcome the variety and genetype restriction in regeneration culture of alfalfa high frequency somatic embryos, vastly improve somatic cell embryo occurrence frequency and regeneration plant frequency, and enhance breeding efficiency.

Description

technical field [0001] The invention relates to a high-frequency somatic embryo regeneration medium of leguminous herbage, more specifically relates to a high-frequency somatic embryo regeneration medium which overcomes the genotype limitation of alfalfa varieties, and belongs to the field of botany. Background technique [0002] Alfalfa (Medicago sativa L.) has been widely planted in my country due to its outstanding economic value and ecological function, and plays an irreplaceable role in the development of agriculture and animal husbandry and the improvement of ecological environment. In the face of production needs and existing production conditions, alfalfa needs to be further improved in terms of yield, quality and resistance. Combining biotechnology with conventional breeding methods to breed new alfalfa varieties is an effective way to improve alfalfa quality and breeding efficiency. [0003] Alfalfa is a perennial cross-pollinated plant with a very low self-seed s...

Claims

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Application Information

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IPC IPC(8): A01H4/00C05G1/00
Inventor 梁慧敏夏阳燕丽萍王小春鲍荣静
Owner JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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