Plant protoplast culturing method

A protoplast and culture method technology, applied in the field of cell biology, can solve problems such as poor repeatability, low plant efficiency, and difficulty in fusion breeding, and achieve the effect of overcoming the bottom of yield and improving efficiency and yield

Pending Publication Date: 2019-07-09
HEFEI NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But until now, plant protoplast fusion breeding is still very difficult, the main reason is that the regeneration eff

Method used

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Examples

Experimental program
Comparison scheme
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Example Embodiment

[0009] Hereinafter, the technical solution of the present invention will be described in detail through specific embodiments.

[0010] 1. Thawing and activation of protoplasts

[0011] (1) Take a centrifuge tube and use a dropper to drop 5-9ml of culture solution into it.

[0012] (2) Take out the freezing tube from the liquid nitrogen tank, and quickly put it in a 36℃-38℃ water bath, shake it from time to time to make it melt rapidly, and it will be completed within 30-60s.

[0013] (3) After the cryopreservation tube is wiped and disinfected with 70% alcohol, the lid is opened, and the protoplast suspension is transferred into the above-mentioned centrifuge tube with a straw.

[0014] (4) Centrifuge at low speed (1200r / min) for 6 minutes, remove the supernatant, and wash and centrifuge again with 5-9ml of culture medium.

[0015] 2. Bottling culture

[0016] (1) Remove the supernatant after centrifugation, add 3ml of culture medium (MS) dropwise to the centrifuge tube, and mix.

[0017] ...

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Abstract

The invention discloses a plant protoplast culturing method. According to the plant protoplast culturing method, in a suspension culture condition, compound sodium nitrophenolate is employed to activate protoplasm and reinforce the intensity of stimulation to the protoplasm, so that in-vitro amplification capacity of the protoplasm is improved; a high-temperature water bath and then a quick ice bath are performed before elution of the protoplasm, so that a low yield rate caused by adhesion of the protoplasm is avoided. The method is easily controllable in culture process, simple and convenientto operate and easy for expansion.

Description

technical field [0001] The invention relates to the field of cell biology, in particular to a plant protoplast culture method. Background technique [0002] Plant protoplast fusion technology is based on the research results of animal cell fusion and established on the basis of protoplast isolation and culture. Using plant protoplasts as materials, the two protoplasts are fused through the induction of physical and chemical factors. together so as to form fused cells. It is not a union between male and female spores, but a fusion between somatic cells with complete genetic material, a hybrid between two kinds of protoplasts. Two cells with different genomes can be combined through protoplast fusion. Compared with conventional sexual hybridization, it can not only expand the range of hybrid parent combinations, but also shorten the breeding cycle and reduce the risk of breeding new varieties. Small. But so far, plant protoplast fusion breeding is still very difficult, the ...

Claims

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Application Information

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IPC IPC(8): C12N5/04
CPCC12N5/04
Inventor 王子迎徐德聪吴林生沈章军
Owner HEFEI NORMAL UNIV
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