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

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...

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 been more than 30 years of alfalfa tissue culture research, and the explants used in domestic and foreign research reports include almost all organs or tissues of alfalfa, such as shoot tips, leaves, stem segments, sprouts, cotyledons, hypocotyls, epicotyls, Anthers, pollen, ovary, ovules, young roots, petioles, etc., the medium is liquid or solid, and a variety of basic mediums have been used. The best explant obtained is the hypocotyl. About alfalfa somatic embryo regeneration plants There have been some reports, but the results obtained in these studies all indicate that the somatic embryogenesis ability of alfalfa is genotype-dependent, so these studies are also difficult to solve the problem of alfalfa genotype barriers

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