High-frequency somatic embryo regeneration culture method for overcoming alfalfa variety genotype limitation

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

Inactive Publication Date: 2010-12-29
JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY +1
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  • Application Information

AI Technical Summary

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...

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  • High-frequency somatic embryo regeneration culture method for overcoming alfalfa variety genotype limitation
  • High-frequency somatic embryo regeneration culture method for overcoming alfalfa variety genotype limitation
  • High-frequency somatic embryo regeneration culture method for overcoming alfalfa variety genotype limitation

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specific Embodiment approach

[0073] 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

[0131] 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

[0133] 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, germination and seedling growth 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 high-frequency somatic embryo regeneration culture method for overcoming alfalfa variety genotype limitation, which comprises the following steps of: (1) selecting materials: selecting three explants of the epicotyl, the hypocotyl and the regenerated plant blade of an aseptic seedling after flushing, sterilizing and disinfecting as well as drying a mature alfalfa seed; (2) inoculating the selected explants onto a somatic embryogenesis induced culture medium for culture; (3) inoculating the explants subjected to somatic embryogenesis induced culture onto an embryo successive formation culture medium for culture; (4) inoculating the explants subjected to embryo successive formation culture onto an embryo maturation germination seeding-formation culture medium for culture to form a seedling; (5) carrying out seedling strengthening and rooting culture; and (6) transplanting a regenerated plant to survive. The invention can effectively overcome the problems of different varieties and genotype limitation in alfalfa high-frequency somatic embryo regeneration culture, greatly improves the somatic embryogenesis frequency and the plant regeneration frequency and improves the breeding efficiency.

Description

technical field [0001] The invention relates to a method for high-frequency somatic embryo regeneration and cultivation of leguminous herbage, and more specifically relates to a method for overcoming the genotype restriction 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 setting rate, less th...

Claims

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Application Information

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IPC IPC(8): A01H4/00C05G3/00
Inventor 梁慧敏夏阳汤荣娣王小春燕丽萍
Owner JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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