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Method for establishing brazilian panama rubber tree suspending cell line

A technology of Hevea brasiliensis and suspension cells, which is applied in the field of establishment of Hevea brasiliensis suspension cell lines, can solve problems such as the inability to obtain a large number of autogenous clone plants, the inability to obtain embryogenic callus at low cost, and the inability to proliferate embryogenic callus. Achieve the effect of low production cost, wide application field and high efficiency

Inactive Publication Date: 2009-12-23
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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Problems solved by technology

However, the main problems in the production of self-rooted clonal plants of Hevea brasiliensis are: the acquisition of anthers and inner integuments is limited by seasons, it is impossible to obtain embryogenic callus in a cheap, large, continuous and long-term manner, and it is impossible to control embryogenic callus. Large-scale multiplication of sexual callus, so it is impossible to obtain a large number of self-rooted clones, and thus unable to meet the demand for a large number of self-rooted clones in field production

Method used

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Embodiment

[0024] A method for setting up Hevea brasiliensis suspension cell line, the concrete steps are:

[0025] A. Taking rubber tree flower buds as explants, the flower buds were surface-sterilized with 75% alcohol for 60 seconds, then sterilized with 0.1% mercuric chloride for 15 minutes, and finally rinsed 3 times with sterile water, 5 minutes each time;

[0026] B. Peel off the stamens in the sterilized flower buds, inoculate them on the induction medium and carry out dark culture at a temperature of 28°C for 50 days; the induction medium is based on MS medium, and MS is basically cultured The content of potassium dihydrogen phosphate, calcium chloride anhydrous, manganese sulfate tetrahydrate and magnesium sulfate heptahydrate in the base is adjusted to potassium dihydrogen phosphate 350mg / L, calcium chloride anhydrous 200mg / L, manganese sulfate tetrahydrate 20mg / L L. Magnesium sulfate heptahydrate 400mg / L, add 2,4-D 2mg / L, KT 1mg / L, NAA 1mg / L, Phytagel 2g / L, sucrose 70g / L, coco...

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Abstract

The invention discloses a method for establishing a suspension cell line of Hevea brasiliensis, which comprises the preparation process of explants, the induction process of embryogenic callus, the subculture process of embryogenic callus, and the suspension culture process of embryogenic callus. The invention has simple technological process, low production cost, rapid and stable multiplication of the established suspension cell line, and can be subcultured for a long time, and solves the problem of conventional Hevea brasiliensis embryogenic callus being affected by seasons and climates, as well as large-scale multiplication and costing a lot The problem of manpower and material resources provides an important foundation for the large-scale commercialization, factory production and large-scale field planting of Hevea brasiliensis root clones, and for the creation of Hevea brasiliensis using biotechnology methods such as somatic cell hybridization, genetic transformation and mutant induction. The new excellent germplasm resources provide new approaches and conditions, laying the foundation for the research and development of Hevea brasiliensis bioreactors.

Description

technical field [0001] The invention belongs to the technical field of plant cell engineering, and more specifically relates to a method for establishing a suspension cell line of Hevea brasiliensis. Background technique [0002] Natural rubber is one of the four major industrial raw materials and an extremely important strategic material. At present, among more than 2,000 kinds of rubber-producing plants in nature, about 99% of commercial natural rubber comes from Hevea brasiliensis. Hevea brasiliensis is a perennial tree belonging to the family Euphorbiaceae, and the conventional breeding cycle is 20 to 30 years. The ultra-long breeding cycle has greatly restricted the selection and renewal of new varieties of Hevea brasiliensis. In order to broaden the breeding approaches and innovate breeding methods of Brasilia, since the 1950s, breeders from all over the world have begun to study the application of biotechnology in Brasilian breeding. The success of somatic embryogen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/02C12N5/04
Inventor 华玉伟黄华孙黄天带吴紫云曾霞胡彦师李维国李哲
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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