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

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...

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

Images