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Method of using Na2S2O3 (sodium thiosulfate) to preventing browning of tobacco explant by using Na2S2O3 (sodium thiosulfate)

A technology of sodium thiosulfate and explants, which is applied in the field of plant tissue culture, can solve the problems that plants cannot continue to be cultivated, plants die, etc., and achieves the effects of relieving browning phenomenon, good effect and improving differentiation state.

Inactive Publication Date: 2018-06-12
CHINA TOBACCO YUNNAN IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the browning problem of tobacco in the tissue culture process is common and almost inevitable. Browning will lead to the death of the plant, that is, the plants in the culture medium cannot continue to be cultivated, so preventing browning is in tobacco tissue culture. problems to be solved
[0005] The present invention utilizes adding anti-browning agent sodium thiosulfate (Na 2 S 2 o 3 ) to alleviate the browning of tobacco explants, and related methods have not been reported

Method used

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  • Method of using Na2S2O3 (sodium thiosulfate) to preventing browning of tobacco explant by using Na2S2O3 (sodium thiosulfate)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Step (1), Disinfection of Tobacco Seeds: Take the plump tobacco seeds, then move them to the ultra-clean workbench, soak the seeds with 75% alcohol for 30s, rinse them twice with sterile water, and then wash them with 1% AgNO 3 Disinfect for 10 minutes, soak and wash in sterile water for 5 times, dry the surface moisture of the explants with sterile filter paper, and then inoculate, wherein the sterile water is ultrapure water sterilized by autoclaving;

[0025] Step (2), culturing tobacco seeds: Inoculate the sterilized tobacco seeds obtained in step (1) into MS medium at a culture temperature of 25±1°C and light intensity of 30-50 μmol / (m 2 s), the light time is 16h / d under the condition of cultivating 60d, and the seeds germinate and grow into seedlings. The MS medium is: the agar of MS basic medium+30g / L sucrose+4g / L, the pH of the medium The value is 5.7;

[0026] Step (3), tobacco explant culture: the seedlings obtained in step (2) are punched with a hole puncher...

Embodiment 2

[0034] Step (1) to step (3) are the same as in Embodiment 1.

[0035] Step (4), anti-browning cultivation of tobacco explants: place the tobacco explants obtained in step (3) in MS primary anti-browning medium, culture at a temperature of 28 / 25°C for 16 / 8 hours, Light intensity 30-50μmol / (m 2 Cultivate under the condition of s) and cultivate respectively for 20d, and the MS primary anti-browning medium is that the anti-browning additive 4.0g / L sodium thiosulfate (Na 2 S 2 o 3 ), 30g / L sucrose, the agar of 4g / L, the pH value of culture medium is 5.7;

[0036] Step (5), subculture anti-browning culture of tobacco explants:

[0037] 1) to 3) are identical with embodiment 1;

[0038] 4) Place the preliminary anti-browning tobacco explants obtained in step (4) in the MS subculture anti-browning medium, and culture at a culture temperature of 28°C for 16h / d in the light and 8h / d in the dark at 25°C , light intensity 30-50μmol / (m 2 Cultivate under the condition of s) for 20 da...

Embodiment 3

[0040] Step (1) to step (3) are the same as in Embodiment 1.

[0041] Step (4), anti-browning cultivation of tobacco explants: place the tobacco explants obtained in step (3) in MS primary anti-browning medium, culture at a temperature of 28 / 25°C for 16 / 8 hours, Light intensity 30-50μmol / (m 2 Cultivate under the condition of s) and cultivate respectively for 20d, and the MS primary anti-browning medium is that adding anti-browning additive 5.0g / L sodium thiosulfate (Na 2 S 2 o 3 ), 30g / L sucrose, the agar of 4g / L, the pH value of culture medium is 5.7;

[0042] Step (5), subculture anti-browning culture of tobacco explants:

[0043] 1) to 3) are identical with embodiment 1;

[0044] 4) Place the preliminary anti-browning tobacco explants obtained in step (4) in the MS subculture anti-browning medium, and culture at a culture temperature of 28°C for 16h / d in the light and 8h / d in the dark at 25°C , light intensity 30-50μmol / (m 2 Cultivate under the condition of s) for 20...

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Abstract

The invention relates to a method using Na2S2O3 (sodium thiosulfate) to of preventing browning of a tobacco explant by using Na2S2O3 (sodium thiosulfate). The method comprises the following steps of disinfection of plant seeds, culturing of the plant seeds, culturing of the plant explant, anti-browning culturing of the plant explant, and subculture type anti-browning culturing of the plant explant, so that the browning rate of the plant explant is 18.7 to 23.3%. The method has the advantages that the browning of the explant is effectively relieved, and the whole differentiation state of the explant can be improved; the Na2S2O3 and natural anti-oxidizing components are added into a culture medium; the convenience in use is realized, the cost is low, the effect is good , and the like.

Description

technical field [0001] The invention belongs to the field of plant tissue culture, and in particular relates to a method for preventing tobacco explants from browning by using sodium thiosulfate. Background technique [0002] Tobacco belongs to the dicotyledonous class in botany ( Dicotycoledoneac ), Tubeflowers ( Tubiflorae ), Solanaceae ( Solanaceae ), Nicotiana, Nicotiana is the fifth largest family of Solanaceae plants. Currently Nicotiana species except common tobacco ( N. tabacum L. ) and Nicotiana nicotiana are widely used in the tobacco industry, and the rest are classified as wild species of Nicotiana. It has been reported that tobacco is often used in the research of basic biochemical metabolism, genome evolution, transgenic and other fields, and common tobacco, as one of the model organisms for studying plants, is an important tool commonly used in the field of plant molecular research. [0003] Browning of explants in the process of plant tissue culture i...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/008A01H4/001
Inventor 许力向海英蒋佳芮邓乐乐张建铎曾婉俐高茜宋春满张涛李雪梅陈章玉
Owner CHINA TOBACCO YUNNAN IND
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