Method for obtaining transgenic cotton through transformation of live immature embryos mediated by Agrobacterium

An Agrobacterium-mediated, living immature embryo technology, applied in biochemical equipment and methods, horticultural methods, genetic engineering, etc., can solve the problem of low multi-copy integration probability of foreign genes, different cotton regeneration ability, and long tissue growth cycle. problems, to achieve the effect of overcoming the long growth cycle, overcoming the limitations of cotton lines, and short growth cycles
CN104726489BInactive Publication Date: 2018-07-10SHAANXI NORMAL UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
SHAANXI NORMAL UNIV
Publication Date
2018-07-10
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a method for obtaining transgenic cotton by transforming live immature embryos mediated by Agrobacterium, which consists of the steps of peeling off aseptic embryos, preparing transformation liquid, shaking and infecting, preparing transgenic cotton, and identifying transgenic plants. In the aseptic embryo stripping, 30% hydrogen peroxide and sterile water are prepared into a disinfectant solution, the cotton seeds are plucked and put into the disinfectant solution, soaked at 25°C for 24 hours, the seeds are taken out and placed in a petri dish, and the seed coat and 1 / 3 to 1 / 2 cotyledons, and the remaining cotton embryos are placed in a petri dish containing basal medium to injure the cotton embryos; in the step of preparing the transformation solution, the induction solution is made of acetosyringone and magnesium sulfate, and acetosyringone Ketones can induce the activation of the Vir gene of Agrobacterium and improve the efficiency of exogenous gene integration; in the shaking infection step, Silwet L‑77 and MS liquid medium were added to the transformation solution to make Agrobacterium cover cotton embryos in a wider range. The invention has the advantages of easy screening, short cotton growth cycle, easy operation, transformation efficiency and the like.
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Description

technical field

[0001] The invention belongs to the technical field of transgenic cotton, and in particular relates to the establishment of a method for obtaining transgenic cotton by transforming live immature embryos mediated by Agrobacterium. Background technique

[0002] Cotton is an important fiber crop, and its fiber production accounts for 40% of the world's total fiber production, and the planting amount of upland cotton reaches 90% of the total cotton. Cottonseed is an oil-containing plant seed second only to soybeans and rapeseeds, so cotton plays an important role in economic crops.

[0003] Improving the quality of cotton fiber and increasing the oil yield of cottonseed through transgenic technology are important means for cultivating new cotton varieties. In 1987, transgenic cotton plants were obtained for the first time by Agrobacterium-infected cotyledon hypocotyl transformation method, followed by particle bombardment embryo suspension cell method and pollen...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
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