Rubber tree protolast culture plant regeneration method

A protoplast and rubber tree technology, applied in the field of plant tissue and cell engineering, can solve the problems of rubber tree protoplast regeneration difficulties, difficulty in obtaining, complicated preparation procedures, etc., to achieve continuous division and further growth and development, simplification of ingredients and preparation process , The effect of the system is simple and easy to implement

Inactive Publication Date: 2014-12-17
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Foreign scholars took the lead in the research on the isolation and cultivation of rubber tree protoplasts in the 1980s. However, because the regeneration of rubber tree protoplasts is quite difficult, and there are great differences in the responses of different varieties to the same culture conditions, many scholars are in this research. failed in the field
Until 2000, Sushamakumari et al first reported that the protoplast culture of the rubber tree variety PRII105 successfully obtained complete regeneration plants, but there have been no successful reports of other varieties since then.
The reason may be that this culture system is not suitable for other species of rubber tree, especially the protoplast regeneration culture of domestic rubber tree species, and the culture system has the following deficiencies: (1) The KPR medium used for protoplast culture (Abdullah et al. , 1986) complex ingredients, containing a variety of rare and expensive vitamins, high experimental costs and complicated preparation procedures; (2) using Poaceae ryegrass suspension cells as care cells, which are not easy to obtain, and the protoplasts of rubber trees after care and culture divide to form Cell clones are less frequent
In view of the above-mentioned technical bottlenecks in the cultivation and regeneration of rubber tree protoplasts, there is no relevant research report in this field in my country at present.

Method used

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Embodiment Construction

[0027] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0028] When a kind of rubber tree protoplast of the present invention cultivates the method for plant regeneration to implement, step is as follows:

[0029] (1) Establishment of embryogenic cell suspension line: Pick 3g of embryogenic callus induced by immature anthers of rubber tree, transfer it into a 100ml Erlenmeyer flask containing 30ml of liquid medium, place it on a shaker at 100rpm, and keep it at 28°C in the dark Suspension culture was carried out under the following conditions, subcultured once a week for 80 days, and a stable and homogeneous suspension cell line was obtained.

[0030] The composition of the liquid medium is: improved MS basal medium, 1mg L -1 6-BA, 1mg·L -1 NAA, 1.5mg·L -1 2,4-D, 0.2g·L -1 Asparagine, 0.4g·L -1 Acid Hydrolyzed Casein, 75ml·L -1 Coconut water and 50g·L -1 Sucrose; liquid medium pH 5.8...

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Abstract

The invention discloses a rubber tree protolast culture plant regeneration method which comprises the following steps of: establishing an embryonic cell suspension system, separating and purifying protoplast, nursing and culturing the protoplast, carrying out mulitiplication in protoplast regeneration cell clone, inducing somatic embryos and regenerating plants. The rubber tree protolast culture plant regeneration method disclosed by the invention is simple and easy to carry out, the components of a nursing and culture medium employed to nursing and culturing the protoplast are greatly simplified, and the suspension cell systems of the varieties of the rubber tree self are used as the nursing cells, so that the production cost is greatly lowered, the efficiency that the rubber tree protolast is split to form cell clone and the efficiency that the somatic embryos are generated subsequently are effectively improved. A good receptor system is provided for rubber tree variety improvement and quality innovation by utilizing biotechnological breeding modes with significant application prospects, such as protoplast fusion (namely somatic hybridization) and genetic transformation.

Description

technical field [0001] The invention belongs to the field of plant tissue and cell engineering, in particular to a method for cultivating and regenerating rubber tree protoplasts. Background technique [0002] Hevea brasiliensis Müll.Arg. (hereinafter referred to as rubber tree) is an important tropical economic crop, and the natural rubber it produces is an irreplaceable industrial raw material and national defense strategic material. With the development of the national economy, my country has become the world's largest consumer of natural rubber, with an annual demand of more than 3.8 million tons. However, my country's current annual output does not exceed 800,000 tons, and the self-sufficiency rate is only about 20%, which is far from reaching the international level. Security cordon, if effective measures are not taken as soon as possible to increase the output of natural rubber, it will directly threaten the country's economic development and national defense security....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 戴雪梅周权男黄华孙华玉伟李哲黄天带孙爱花
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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