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Agrobacterium mediated method for obtaining transgenic plant of hevea brasiliensis

An Agrobacterium-mediated, rubber tree technology is applied in the field of plant genetic engineering to achieve the effects of fast growth, high transformation efficiency and great application value

Inactive Publication Date: 2012-09-19
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limitations of various factors, there are still many difficulties in establishing an efficient genetic transformation system. How to solve these difficulties and finally establish a stable and efficient genetic transformation system has become the most urgent task at present.

Method used

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Examples

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Embodiment Construction

[0019] The following examples are intended to illustrate the present invention, but not to limit the scope of the present invention.

[0020] Various culture media and engineering bacteria liquids were prepared in advance as required. Take the strain of Agrobacterium tumefaciens EHA105 (containing plasmid pCambia2301-AtCBF1) stored at -80℃, streak on YEB solid medium containing 50mg / L kanamycin and 50mg / L rifampicin, and invert at 28℃ for 60h , pick a single colony and place it in YEB liquid medium supplemented with the same antibiotics, shake and culture at 220rpm for 30h at 28°C until Agrobacterium tumefaciens is in the logarithmic growth phase, then take 20ml of bacterial liquid, centrifuge with sterile 50ml The tube was centrifuged at room temperature 4000rpm for 10min to collect the thalline, the collected thalline was resuspended with 100ml of AAM medium, and after shaking cultured for 5h under the same conditions, it was diluted with fresh AAM medium to OD600=0.4 (OD600...

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Abstract

The invention relates to an agrobacterium mediated method for obtaining a transgenic plant of hevea brasiliensis. The method comprises the following steps of: preparing an engineering bacteria liquid, infecting embryogenic suspension cells of hevea brasiliensis with engineering bacteria, co-culturing the embryogenic suspension cells of hevea brasiliensis with the engineering bacteria, removing agrobacterium from the embryogenic suspension cells of hevea brasiliensis and generating embryogenic calli through induction, screening and subculturing the resistant embryogenic calli, inducing an embryoid with the resistant embryogenic calli and culturing the mature embryoid to obtain a regeneration plant. The transgenic plant can be a fine variety with high hevea brasiliensis cultivation yield, strong resistance and high growth speed, has high transformation efficiency and high regeneration transgenic plant obtainment rate and has great application value.

Description

technical field [0001] The invention relates to an Agrobacterium-mediated method for obtaining transgenic plants of rubber trees, belonging to the technical field of plant genetic engineering. Background technique [0002] Rubber tree (Hevea Brasiliensis Mull.Arg) belongs to the genus Hevea of ​​Euphorbiaceae and is a perennial cross-pollinating tree cultivated in tropical regions. Due to the long life cycle of rubber trees, it takes more than 6 years from the germination of seeds to the cutting of young rubber trees, so the conventional breeding cycle is longer, generally about 30 years. The genetic modification of Brazilian rubber tree by transgenic technology can create excellent conditions for the improvement of rubber tree varieties and shorten the breeding cycle. At present, many countries have invested a lot of manpower and material resources to conduct extensive research and have made some progress (Leclercq J, Martin F, Lardet L, et al. Genetic transformation and r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H4/00A01H5/00
Inventor 周权男姜泽海李哲黄华孙黄天带戴雪梅谢黎黎毕政鸿
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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