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Method for inducing litsea cubeta multiple shoot

A technology of litsea cubeba and explants, which is applied in the field of rapid propagation of tissue culture for inducing clustered buds of woody aromatic plant litsea cubeba, can solve the problem of low seedling rate, unstable survival rate, and tissue culture of litsea cubeba seedlings Insufficient rate stability and other issues, achieve stable genetic traits, improve disinfection effect, and inhibit browning of medium and explants

Inactive Publication Date: 2008-03-26
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the existing tissue culture method can obtain a higher multiplication coefficient, it has the problems of low seedling yield and unstable survival rate after transplanting.
Therefore, it is necessary to explore and study a method of inducing clustered bud culture that can not only guarantee a certain multiplication coefficient but also ensure an effective number of sprouts, so as to solve the key problem of unstable seedling rate of Litsea Cubeba in tissue culture

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Get the young and tender branches of Litsea Cubeba dry base, remove the leaves, put in 3% H2O after washing with running water. 2 o 2 Soak in medium for 15 minutes, take it out and immediately transfer to 0.1% HgCl containing 5% Tween-80 2 Soak in the solution for 20 minutes, constantly brush the branches with a soft brush, then take them out and rinse them with sterile water for 5 to 6 times and dry the water with sterile absorbent paper, then cut out the stems containing nodes as explants, and quickly inoculate them on the The explants were cultured in the dark on MS medium, and the explants were transferred to new MS medium the next day.

[0021] The explants that were no longer browned on the MS medium were transferred to MS+6-BA1.0mg / L+IBA0.5mg / L+GA2.0mg / L+agar 7.0g / L+sucrose 30g / L, pH 6.0 On the induction medium, place at a temperature of 25±2°C and a photoperiod of 12h.d -1 , Induction culture was carried out in an incubator with a light intensity of 1200 lux....

Embodiment 2

[0025] Get the young and tender branches of Litsea Cubeba dry base, remove the leaves, put in 3% H2O after washing with running water. 2 o 2 Soak in medium for 20 minutes, take it out and immediately transfer to 0.1% HgCl containing 5% Tween-80 2 Soak in the solution for 10 minutes, constantly brush the branches with a soft brush, then take them out and rinse them with sterile water for 5 to 6 times and dry the water with sterile absorbent paper, then cut out the stems containing nodes and inoculate them on MS medium, put Culture in the dark, and transfer to new MS medium the next day.

[0026] The explants that were no longer browned on the MS medium were transferred to MS+6-BA0.5mg / L+IBA0.2mg / L+GA1.0mg / L+agar 7.0g / L+sucrose 30g / L, pH 6.0 On the culture medium, place at a temperature of 25±2°C and a photoperiod of 12h.d -1 , Induced culture was carried out in an incubator with a light intensity of 1200 lux. After 21 days, induced shoots were obtained.

[0027] After 30 d...

Embodiment 3

[0030] According to the method of Example 1, only the hormone concentration in the subculture stage was reduced. Specific method: After 30 days of multiplication and culture, cut out the section-containing stem section of the bud and inoculate it into MS+6-BA1.0mg / L+IBA0.3mg / L+GA1.0mg / L agar 7.0g / L+sucrose 30g / L, pH6. 0 medium; after 30 days, transfer the buds to MS+6-BA2.0mg / L+IBA0.3mg / L+agar 7.0g / L+sucrose 30g / L pH 6.0 medium in the same way for subculture. The temperature is 25±2°C, the photoperiod is 14h.d-1, and the light intensity is 2000 luX; such alternate culture, the multiplication coefficient is more than 4 times, and the sprouts obtained are strong and neat.

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Abstract

The present invention relates to a method for inducing litsea cubeba tufted bud. Said method includes the following steps: adopting joint-containing stem part of litsea cubeba as explant, adopting a solution prepared by using 3% of H2O2 and 0.1% of HgCl2 containing 1-5% of Tween-80 and soaking said explant in said above-mentioned solution, and continuously brushing said explant by using soft hair brush to make disinfection instead of ethyl alcohol, at the same time of making high-effective sterilization ensuring enough effective number of explants; in the primary stage of inoculation adopting culture medium and dark culture conversion method to control browning of material; in the bud induction culture stage adding GA; in the bud enrichment culture stage adopting the method for alternatively using enrichment culture medium and induction culture medium; bud germination time is moved up and can be quickly come into the enrichment state, so that the obtained buds are strong and even.

Description

technical field [0001] The invention relates to a method for inducing clustered buds of litsea cubeba, in particular to a tissue culture rapid propagation method for inducing clustered buds of woody aromatic plant litsea cubeba. Background technique [0002] Litseacubeba is the collective name of many plants in the genus Zingiberaceae of Lauraceae. There are more than 200 species in the world, and 72 species, 18 varieties and 3 variants in my country, mainly distributed in the south of the Yangtze River. Its aliases are Litsea jasmine (Zhejiang), Litsea cypress (Guangdong), Sai camphor tree (Fujian), Fragrant leaves (Hunan), and Mujiangzi (Guangxi). The volatile aromatic oil obtained by distillation is called litsea cubeba oil, which is a bulk essential oil for export exchange. Litsea cubeba essential oil is mainly used to extract citral, with a content of 70%. It is the main raw material for the manufacture of ionone, methyl ionone, ethyl ionone and vitamin A, and is used ...

Claims

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Application Information

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IPC IPC(8): A01H4/00C12N5/04
Inventor 张艳玲姚雷王维鹏张静懿高远陆伟芳陆燕李昂
Owner SHANGHAI JIAO TONG UNIV
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