Method for establishing rice suspension cell line and preparation process thereof

A suspension cell and rice technology, which is applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of long duration and shortening the process of suspension cell culture.

Inactive Publication Date: 2018-11-13
江西省超级水稻研究发展中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] The present invention aims at the problem that the existing suspension cell culture procedure of rice takes a long time, and provides a method for establishing a rice suspension cell line, which can shorten the suspension cell culture process, thereby improving the embryonicity of the suspension cell line and reducing clonal variation

Method used

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  • Method for establishing rice suspension cell line and preparation process thereof
  • Method for establishing rice suspension cell line and preparation process thereof
  • Method for establishing rice suspension cell line and preparation process thereof

Examples

Experimental program
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Embodiment 1

[0046] The establishment of embodiment 1 rice variety Kasalath suspension cell line

[0047] The Kasalath suspension cell line was obtained by the following steps:

[0048] 1. Acquisition of embryogenic callus for inoculation

[0049]The mature seeds of Kasalath were used as explants for callus induction, and the method of Toki et al [Toki S, Hara N, Ono K, et al..Plant J, 2006,47(6):969-976] Induction, the medium was N6D solid medium, and the callus was excised and subcultured in the same medium after 1 week. In subsequent subcultures (1 time every 2 weeks), attention should be paid to the combination of loose and friable, hard texture, and granules. The thin embryogenic callus was used for suspension cell culture after subculture for 2 months, and the fresh embryogenic callus grown on fresh medium (N6D solid medium) for 10-12 days was selected for suspension culture.

[0050] 2. Suspension cell culture

[0051] 1) 3-5g rice embryogenic callus is inoculated in 30ml NBL liq...

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Abstract

The invention discloses a method for establishing a rice suspension cell line. The method includes the following steps: (1) obtaining an embryogenic callus for inoculation; (2) inoculating the obtained rice embryogenic callus into a medium and conducting shaking culture in a shaker; (3) filtering out large particles in the culture, collecting a suspension and centrifuging to collect a precipitate;(4) adding a fresh medium, conducting suspension culture on the collected precipitate, conducting subculture for many times until small particle cell masses at the bottom of the medium are produced in large quantities and a culture solution becomes clearer; (5) absorbing small particles in the culture solution for subculture, adding the medium, transferring the small particles into a different medium for subculture after the subculture is conducted twice, and absorbing small particles for subculture for several several days to obtain the rice suspension cell line. By means of the method for establishing the rice suspension cell line, the suspension cell culture process can be shortened, the embryogenicity of the suspension cell line can be improved, and the clonal variation is reduced.

Description

technical field [0001] The invention belongs to the technical field of rice tissue cell culture, and in particular relates to a method for establishing a rice suspension cell line. Background technique [0002] Suspension cell culture is an important plant biotechnology, and the suspension cell line obtained by suspension culture is an important material for plant physiology and biochemistry research, protoplast isolation and bioengineering. [0003] In rice, the establishment of embryogenic suspension cell lines generally goes through the following steps: [0004] First is the preparation of callus, which requires that the callus used for inoculation should be loosely combined, have smaller particles and a harder texture, and have regeneration (embryogenicity); [0005] The second step is preliminary suspension culture. A certain amount of small particle callus is obtained after suspension culture for about one month after inoculation. When subculture, a part of the old cu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005
Inventor 龙起樟万建林黄永兰唐秀英王会民芦明
Owner 江西省超级水稻研究发展中心
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