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Method for converting island cotton by agrobacterium-mediated transformation

An Agrobacterium-mediated, Agrobacterium-mediated technology, applied in the field of plant genetic engineering, can solve the problems of low frequency of cell regeneration, unstable conditions, unsuitable for large-scale production of transgenic plants, etc., and achieve the effect of improving the efficiency of genetic transformation

Inactive Publication Date: 2012-01-04
陈全家 +1
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Problems solved by technology

The successful report on the regeneration system of sea-island cotton was not made public until 2001. The regeneration frequency of somatic cells in this system is low, and the seedling cycle is long. Cell suspension culture is used, and the unstable conditions are not suitable for large-scale production of transgenic plants.

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  • Method for converting island cotton by agrobacterium-mediated transformation
  • Method for converting island cotton by agrobacterium-mediated transformation

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Embodiment Construction

[0020] The method for Agrobacterium-mediated transformation of Gossypium sea-islands, comprising the following steps:

[0021] 1) Receptor culture: Disinfect the surface of sea island cotton seeds. The disinfection method is to first soak in 75% alcohol for 30 seconds, remove the alcohol and soak in 10% hydrogen peroxide for 4 hours, and then use sterilized Rinse with distilled water 4-5 times repeatedly, and finally add appropriate sterile distilled water to soak overnight. The surface-sterilized seeds were inoculated on 1 / 2 MS medium for cultivation. The cultivation conditions were 28°C, three days of dark cultivation, and then 8 hours of 2000 LX light and 16 hours of darkness for 4 days. The hypocotyls obtained from the aseptic vaccines were cut into 0.5-1cm segments with a sharp scalpel, and the incision should not be too large, otherwise it would cause browning and pollution.

[0022] 2) Cultivation of Agrobacterium: pick a single colony, culture it overnight in YEB liqu...

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Abstract

The invention provides an agrobacterium-mediated transformation method of G.barbadense L., which comprises the following steps: (1) receptor culture; (2) agrobacterium culture; (3) the co-culture of the receptor and the agrobacterium with a plant expression carrier; (4) resistant callus induction; (5) resistant cell embryo division; (6) body cell embryo division and resistant regeneration. In theinvention, a regeneration plant of the agrobacterium-mediated transformation of G.barbadense L., is obtained to induce the resistant callus and resistant cell embryo with a hypocotyledonary axis directly serving as the transformation receptor with up to 30 percent of body cell embryo rate and 20 percent of body cell planting rate. The method improves the inheritance transformation efficiency of G.barbadense L. so that the prior biotechnology can be largely applied to G.barbadense L. breeding, solves the key problem of the application of the agrobacterium-mediated method to the G.barbadense L., and provides a technical theory for the G.barbadense L. inheritance improvement.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to a method for Agrobacterium-mediated transformation of sea-island cotton. Background technique [0002] Cotton is an important fiber crop in the world, and it is an urgent problem to be solved in cotton production to efficiently and quickly cultivate excellent cotton varieties with various excellent traits. The application of modern biotechnology in plant breeding is mostly successful on the basis of plant tissue culture technology. Compared with other plants, cotton tissue culture work was carried out later. Cotton wild species Klauski cotton was the first to achieve embryogenesis, but no regenerated plants could be obtained at that time. In 1983, Davidonis et al obtained regenerated plants with the embryogenic callus of Kezi 310 of upland cotton induced for two years. Other investigators have also reported successful embryogenesis and plantlet regeneration. Umbeck et...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N15/87C12N5/10A01H4/00
Inventor 陈全家曲延英贺雅婷孔庆平
Owner 陈全家
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