A kind of efficient and stable broad-spectrum alfalfa genetic transformation method

Abstract

The invention discloses a high-efficiency stable wide-spectrum alfalfa genetic transformation method, and a transformation system obtained by using the method. The method comprises the following steps: 1, preparing a sterilization material; 2, preparing Agrobacterium Tumefaciens; 3, carrying out invasion; and 4, culturing tissues. The Agrobacterium Tumefaciens mediated alfalfa high-efficiency genetic transformation system is established in the invention, lays foundation for advanced research of a transgenic technology and excavation of the gene function of alfalfa, and creates conditions for molecule improvement of the alfalfa. Seven species are compared under the same background (same conversion vector, same insert gene and same explant number), molecular genetic transformation researches of representative species (northeast species, north species and northwest species in China and introduced representative species) of the alfalfa are scientifically and accurately carried out, and research results can represent the conditions of most alfalfa species in China, so the high-efficiency stable wide-spectrum alfalfa genetic transformation method is established.

Description

technical field [0001] The invention relates to the field of genetic transformation of alfalfa, in particular to a method for genetic transformation of alfalfa. Background technique [0002] Alfalfa (Medicago sativa) is an excellent legume forage with the largest planting area and the widest distribution in my country and even in the world. Alfalfa has strong adaptability, rich nutrition and good quality. Stems and leaves are rich in protein, minerals, multivitamins and carotene, and it is an important source of protein and mineral feed for various livestock. In addition to being used as feed, the root nodules of alfalfa also provide nitrogen sources for the soil and play an important role in improving soil structure and physical and chemical properties. Its well-developed root system and excellent resistance make it an important vegetation for water and soil conservation, windbreak and sand fixation. Known as the "King of Grass". [0003] With the development of modern mo...

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Problems solved by technology

The regeneration frequency of different varieties of alfalfa varies greatly, and there are generally disadvantages such as long regeneration cycle, low regeneration efficiency and poor repeatability (Liu Fang. Establishment of Xinjiang yellow alfalfa regeneration system and Agrobacterium-mediated AK-APR gene transformation[D ].Xinjiang Agricultural University, 2012; Ge Jun. Research on tissue culture regeneration system of alfalfa [D]. Beijing: Beijing Forestry University, 2004: 5-6)

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