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Toona sinensis clustered shoot induction and multiplication method

A technology of bud induction and Chinese toon, applied in the field of plant tissue culture, can solve the problems of adventitious bud multiplication of only 5.5, limitations of maternal age and annual fruiting, increase of production cost, etc., to overcome slow propagation speed, reduce production cost, The effect of improving efficiency

Active Publication Date: 2019-10-15
INST OF FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, in the authorized patent ZL201610947913.3 and the patent application CN107006372A, callus differentiation is used for rapid propagation of Toona sinensis, but the callus induction medium of the above two patents needs to be added with TDZ to improve the ability to form callus, but Adding TDZ obviously increases the production cost, which is not conducive to popularization and application in production practice, and the reproduction efficiency of the above two patents is low, and needs to be further improved
In addition, the patent ZL20161094 uses seeds and leaves as explants, and the source of materials is easily limited by the age of the mother and the size of the year.
Kang Bing et al. also disclose the method of adopting callus-induced bud differentiation to carry out the vegetative propagation of Toona sinensis in the article "Callus Induction and Plant Regeneration of Chinese Toon Clones" ("Journal of Horticultural Science" 2004, the first phase, page 15), However, the multiplication factor of its adventitious buds was only 5.5, and it did not reuse the callus many times

Method used

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  • Toona sinensis clustered shoot induction and multiplication method
  • Toona sinensis clustered shoot induction and multiplication method
  • Toona sinensis clustered shoot induction and multiplication method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1, the selection of explant and disinfection mode

[0030] On April 25, 2018, unlignified branches of Toona sinensis tree were collected from the Chinese Academy of Forestry as explants. Remove the leaves on the branches and cut them to about 4-6cm, rinse with washing powder water for 10min, then rinse with running water for 1h, and drain. In the ultra-clean workbench, treat with 75% alcohol for 30s, wash with sterile water for 3 times, and then use 2% sodium hypochlorite (NaClO 3 ) for 5 min, 8 min, and 10 min respectively, after the treatment, wash with sterile water for 3 times, blot the water, and cut the explant into about 1 cm.

[0031] Conclusion: Under the three disinfection methods, the explants were not polluted and browned. Therefore, for saving time, the explant disinfection can adopt the first method, remove the blade on the branch and cut it into 4-6cm, rinse with washing powder water for 10min, then rinse with running water for 1h, and drain....

Embodiment 2

[0032] Embodiment 2, stem segment induced callus

[0033] The sterilized Chinese toon stems were used as explants. Callus culture medium was selected by adding hormones to MS medium. Hormone uses different concentrations of 6-BA, NAA, 2,4-D and hormone combinations (see Table 1-5 for details). The temperature is 25°C, the humidity is 60%, and the light intensity is 20-60 μmolm -2 the s -1 , The light induction time is 16h / d.

[0034] Table 1. Callus induction of Toona sinensis explants treated with different concentrations of 6-BA

[0035]

[0036] Combining Table 1 and figure 1 It can be seen that when callus is induced by stem segments, callus can be induced by adding 6-BA with a concentration of 0.5-5.0mg / L in the medium, and it can be seen that 6-BA is an essential hormone for inducing callus.

[0037] Table 2. Callus induced by different concentrations of NAA

[0038]

[0039] Combining Table 2 and figure 2 It can be seen that when the NAA concentration is ...

Embodiment 3

[0049] Embodiment 3, cluster bud induction

[0050] When the callus grows to about 2cm*2cm, use MS+6-BA2.0mg / L+NAA0.1mg / L, MS+6-BA2.0mg / L+NAA0.2mg / L and MS+6-BA1 respectively .0mg / L+NAA0.1mg / L three kinds of media induce clustered shoots. The temperature is 250C, the humidity is 60%, and the light intensity is 20~60μmolm -2 the s -1 , The light induction time is 16h / d.

[0051] After 30 days, MS+6-BA2.0mg / L+NAA0.1mg / L had no cluster shoots, and the callus induced by MS+6-BA2.0mg / L+NAA0.2mg / L was more than the cluster shoots induced , the cluster bud rate that MS+6-BA1.0mg / L+NAA0.1mg / L induces is 50% (see Figure 5 ). Such as Figure 5 Shown, adopt MS+NAA0.5mg / L+6-BA 0.5mg / L medium to induce the callus differentiation cluster shoot growth vigor than MS+NAA0.5mg / L+6-BA 1.0mg / L medium induction Callus differentiation clustered shoots are more robust, so the induction medium for inducing callus is further preferably MS+NAA0.5mg / L+6-BA0.5mg / L, and the best differentiation me...

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Abstract

The invention discloses a toona sinensis clustered shoot induction and multiplication method. Annual branches which are not lignified are taken as explants, put on a specific induction medium, and induced under the culture conditions of the temperature of 25 DEG C, the humidity of 60%, the illumination intensity of 20-60 mu mol m<-2>s<-1> and the photoinduction time of 16 h / d, callus is formed, and then the callus is induced on a specific differentiation medium under the same culture conditions to form clustered shoots; when the clustered shoots grow to be 3 cm or longer, the clustered shootsare cut, and dense callus is selected for continued multiplication culture. With the adoption of the method, the callus inductivity of a stem can reach 100%, the induction rate of the clustered shootsis higher than 50%, and the multiplication rate of the clustered shoots is higher than 8 and is up to 26.

Description

technical field [0001] The invention relates to the field of plant tissue culture, in particular to a method for inducing and multiplying clustered buds of Toona sinensis. Background technique [0002] Toona sinensis is a precious fast-growing tree species unique to my country, which is used as both wood and vegetables, and has important economic value in agricultural and forestry production. [0003] In the natural state, Toona sinensis mainly reproduces by root tillers. The advantage is that it grows rapidly and can maintain the excellent traits of the mother tree. The disadvantage is that the limited number cannot meet the needs of large-scale cultivation. Seed sowing and cutting seedling propagation are also commonly used methods of Chinese toon propagation. However, the progeny of seed propagation are prone to separation and degeneration of traits, which greatly reduces the commodity value of Chinese toon, while cutting propagation is limited by materials, and the low ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 杨桂娟麻文俊贾子瑞卢楠欧阳芳群王军辉
Owner INST OF FORESTRY CHINESE ACAD OF FORESTRY
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