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Method for establishing hevea brasiliensis suspension cell line

A suspension cell and rubber tree technology, which is applied to plant cells, horticultural methods, botany equipment and methods, etc., can solve the problems of time-consuming and laborious, increased risk of cell line variation, and decreased differentiation ability, so as to reduce production costs and shorten production time. Experimental period, method is simple and easy to implement

Inactive Publication Date: 2016-05-11
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current method for establishing the suspension cell line of Hevea nigra is to induce callus from anther or inner integument solid culture, and after multiple subcultures to screen out fragile embryogenic callus, it is then transferred to liquid medium for suspension Cultivation, at this stage, it still needs to go through multiple subcultures to finally obtain a stable and dispersed suspension cell line, and it takes 6-10 months or even longer from the initial culture of explants to the final establishment of the cell suspension line time, quite time-consuming
The long experimental cycle in the early stage is also very unfavorable for the acquisition of subsequent experimental materials, thus limiting the development of some valuable scientific research
In addition, the differentiation ability of the suspension cell line gradually decreases or even disappears with the increase of the number of subcultures after establishment, and the risk of cell line variation increases

Method used

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Comparison scheme
Effect test

Embodiment 1

[0017] When the method for establishing a rubber tree suspension cell line of the present invention is implemented, the steps are as follows:

[0018] (1) The selection and disinfection of materials is to pick the immature male flower buds of rubber tree at the side-side stage before nine o'clock in the morning, and disinfect the surface of the buds with 75% alcohol for 60 seconds, and then use 0.1% mercury chloride for 10 minutes. After rinsing with sterile water for 4 times, the flower buds were collected with sterilized metal mesh and placed in a petri dish covered with sterile filter paper.

[0019] (2) The process of inducing callus by suspension culture is to completely peel off the anthers in the sterilized flower buds with tweezers and dissecting needles on a clean workbench, and inoculate 60 anthers in a triangular flask containing 15 ml of liquid medium. Cultivate in the dark at a temperature of 27°C on a 90 rpm shaker for 30 days, changing the liquid medium once during t...

Embodiment 2

[0024] When the method for establishing a rubber tree suspension cell line of the present invention is implemented, the steps are as follows:

[0025] (1) The selection and disinfection of materials is to pick out the immature male flower buds of rubber tree at the side-side stage before nine o'clock in the morning, first use 75% alcohol to disinfect the surface of the flower buds for 50 seconds, and then use 0.1% mercury chloride for 12 minutes After rinsing with sterile water for 4 times, the flower buds were collected with sterilized metal mesh and placed in a petri dish covered with sterile filter paper.

[0026] (2) The process of inducing callus by suspension culture is to completely peel off the anthers in the sterilized flower buds with tweezers and dissecting needles on an ultra-clean workbench, inoculate 80 anthers in a triangular flask containing 20 ml of liquid medium, and place Cultivate in the dark at a temperature of 27°C on a 90 rpm shaker for 30 days, changing the ...

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Abstract

The invention discloses a method for establishing a hevea brasiliensis suspension cell line. The method comprises material selection and disinfection, suspension culture induction process for the callus tissue and a subculture screening culture process. The method for rapidly establishing the hevea brasiliensis suspension cell line, which is provided by the invention, is simple in technological process and relatively short in time consumption, is capable of establishing a stable, uniform, good-dispersity and rapid-growth suspension cell line within only 60-90 days which is reduced by at least a half in comparison with that spent in establishing the hevea brasiliensis suspension cell line by use of a conventional method, greatly lowering the production cost, shortening the experiment period and rapidly providing favorable material sources for technical systems of synchronously inducing regeneration plants from a batch of somatic embryos, or performing protolast culture and somatic hybridization and performing direct conversion on foreign genes by utilizing protoplast as a receptor, and the like.

Description

Technical field [0001] The invention belongs to the field of plant tissue and cell engineering, and specifically relates to a method for establishing a rubber tree suspension cell line. Background technique [0002] A good plant suspension cell line is not only the best material for the simultaneous occurrence of a large number of somatic embryos, but also the main source of protoplasts in many crops. After enzymolysis, the cell wall is explained and exposed single cells surrounded by plasma membranes can be introduced as exogenous genes. The direct recipients of this species can also carry out somatic cell hybridization between species and even between species, thereby creating new germplasm resources. In addition, plant suspension cells can also be used to make artificial seeds, screen mutants, and produce secondary metabolites. Therefore, how to establish a good plant suspension cell culture system is a research topic with very practical value and application prospects. [000...

Claims

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Application Information

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IPC IPC(8): A01H4/00C12N5/04
CPCA01H4/00A01H4/001C12N5/04
Inventor 戴雪梅杨先锋黄天带李季辛士超黄华孙
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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