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Method of quickly obtaining transgenic cotton plant

A transgenic, cotton technology, applied in the field of plant molecular biology, can solve the problems of low transformation efficiency, low frequency of occupancy, and long transformation time, and achieve the effects of short cycle, simple operation and high transformation efficiency.

Inactive Publication Date: 2005-01-26
HUAZHONG AGRI UNIV
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Problems solved by technology

The biggest problem of this method is that the conversion takes a long time (at least one and a half years), and the conversion efficiency is low
Although Gould's shoot tip culture method can obtain transformants quickly, the degree of chimerism of transgenic plants is very high, and the frequency of transgenic seedlings in the entire regenerated plant population is very low, so the efficiency of obtaining transgenic plants is not very high.

Method used

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  • Method of quickly obtaining transgenic cotton plant
  • Method of quickly obtaining transgenic cotton plant
  • Method of quickly obtaining transgenic cotton plant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] First soak the shelled cotton seeds with 0.1% mercury chloride for 8-10 minutes, then soak them with 70% alcohol for 3 minutes to disinfect the cotton seeds in the experiment, and then place the sterilized cotton seeds in MS medium ( Add sucrose 20g / L) on aseptic germination, prepare hypocotyl section. The medium used for co-cultivation and selection was commercial MS medium, supplemented with vitamin components of B5 medium, niacin 1mg / L, vitamin B1 20mg / L, vitamin B6 2mg / L, and glucose 30g / L. For the antibiotic used in the medium for selecting transformants, if it is kanamycin, the concentration used is 10-50 mg / L; if it is hygromycin, the concentration is 5 international units / L. For rapid identification of transformants, if the reporter gene is the GFP gene, it can be directly observed with a fluorescent stereo microscope; if it is the GUS gene, it can be observed by conventional methods (see literature: Cell: A Laboratory manual, D.L. Spector et al. 1997). Rooting...

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Abstract

A method for quickly obtaining transgenic cotton plant includes such steps as improving MSB5 culture medium by adding VB1, VB6, nicotinic acid and auximone, culturing the hypocotyl dissection of aseptic cotton, and direct induction. Its advantages are simple and feasible method and high transgenic efficiency (60-70%).

Description

technical field [0001] The invention belongs to the technical field of plant molecular biology and is related to plant transgenic technology. Background technique [0002] Cotton is an important crop that is difficult to use Agrobacterium-mediated transformation to obtain transgenic plants. Although many laboratories at home and abroad have reported examples of cotton transformation using Agrobacterium, it is difficult for the crop to pass through the callus. Plants are differentiated again, so the frequency of successful transformation is extremely low, and even transgenic plants cannot be obtained at all. (P.Umbeck, genetically transformed cotton plants, Bio / Technology, 1987, 5 volumes, 3 phases, 263-266 pages) methods and the shoot tip transformation method such as J.H.Gould etc. (J. H. Gould et al. 1999, Plant Mol. Bio. Rptr. 16:3). The former adopts the process of inducing callus, then inducing embryoid body formation, and finally inducing transgenic plants. The bigg...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 杨业华王学奎
Owner HUAZHONG AGRI UNIV
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