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Agriculture bacillus mediated alfalfa genetic conversion method

A genetic transformation method, Agrobacterium-mediated technology, applied to other methods of inserting foreign genetic materials, horticultural methods, botanical equipment and methods, etc., can solve the problem of complex somatic embryo culture procedures, inhibition of somatic embryo development, and difficulty in mastering, etc. problem, to achieve the effect of improving the efficiency of genetic transformation, reducing the concentration of antibiotics, and shortening the induction cycle

Inactive Publication Date: 2011-05-25
NORTHEAST NORMAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

However, the somatic embryogenesis pathway also has its certain disadvantages, such as the long cycle of the somatic embryogenesis pathway, complex somatic embryo culture procedures, difficult to master, uneven growth status of somatic embryos, and high concentrations of antibiotics severely inhibit the development of somatic embryos, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] A method for genetic transformation of alfalfa hypocotyls mediated by ultrasound-assisted Agrobacterium

[0019] 1 Material:

[0020] 1) Alfalfa seeds are Gongzhuling No. 1, purchased from Jilin Academy of Agricultural Sciences.

[0021] 2) Strains and plasmids: Agrobacterium strain LBA4404 and transformation vector pBI121 were purchased from Beijing Dingguo Biotechnology Co., Ltd.; containing the target gene SsNHX1 (Na + / H + Antiporter gene) transformation vector pBI121 was preserved and constructed by the College of Life Sciences, Northeast Normal University, and is available publicly. The accession numbers of the target gene and the transformation vector pBI121 in the US NCBI database are AF370358 and AF485783, respectively.

[0022] 2 methods:

[0023] 1) Alfalfa seeds should first be polished 5-6 times with fine sandpaper, then soaked in 75% alcohol for 10 minutes, rinsed with sterile water for 3-4 times after pouring off the alcohol, then treated with 0.1% me...

Embodiment 2

[0038] Agrobacterium-mediated Genetic Transformation Method of Alfalfa True Leaf

[0039] 1 Material:

[0040] 1) The alfalfa seed Gongzhuling No. 1 was purchased from Jilin Academy of Agricultural Sciences.

[0041] 2) Strains and plasmids: Agrobacterium strain LBA4404 and transformation vector pBI121 were purchased from Beijing Dingguo Biotechnology Co., Ltd.; containing the target gene SsNHX1 (Na + / H + Antiporter gene) transformation vector pBI121 was preserved and constructed by the College of Life Sciences, Northeast Normal University, and is available publicly. The accession numbers of the target gene and the transformation vector pBI121 in the US NCBI database are AF370358 and AF485783, respectively.

[0042] 2 methods:

[0043] 1) Alfalfa seeds should first be polished 5-6 times with fine sandpaper, then soaked in 75% alcohol for 10 minutes, rinsed with sterile water for 3-4 times after pouring off the alcohol, then treated with 0.1% mercury liter for 20 minutes, po...

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Abstract

The invention discloses an alfalfa genetic inverting method in plant transferring gene technical domain, which comprises the following steps: 1, comprising two methods; evoking callus with alfalfa hypocotyls through floating culture mode; assisting by super sound; sieving body embryo in medium; germinating the body embryo; generating root; getting the transforming plant; 2, using tender true leafof alfalfa as transferring acceptor material; transforming with agricillin medium guild; sieving in traumatic evoked medium; seeding resistance callus in differential medium; evoking out of body embryo; generating root; getting the transforming plant. This invention increases genetic transferring efficiency of alfalfa and shorten genetic transferring period.

Description

technical field [0001] The invention belongs to the field of plant transgenic technology, in particular to a method for genetic transformation of alfalfa Background technique [0002] Although the genetic transformation research of alfalfa has been carried out for more than 20 years, the establishment of an efficient and stable genetic transformation system has always been an important factor restricting the research and application of transgenic alfalfa. In the genetic transformation of alfalfa, somatic embryogenesis is the main pathway of alfalfa regeneration. At present, the reported transgenic alfalfa is mainly obtained through this approach. For example, Mckersied et al. (1996) applied the Mn-SOD gene to transform alfalfa through somatic embryo transformation to obtain drought-tolerant alfalfa. Winicov et al. (1999) successfully integrated the Alfin1 transcription factor into the alfalfa genome using the somatic embryo transformation approach, and obtained salt-alkali-...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N15/87A01H4/00
Inventor 刘立侠李望丰王德利贾广和金太成申玉华常青马强殷东旭许守民
Owner NORTHEAST NORMAL UNIVERSITY
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