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A high-efficiency regeneration method using yellow beam leaves as explants

A technology of explants and yellow beam wood, applied in the field of plant tissue culture, can solve the problems of inaccurate verification of gene function, inconvenient operation, slow propagation speed, etc., to promote industrial development, facilitate regeneration, and facilitate material extraction Effect

Active Publication Date: 2019-01-08
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The existing reports at home and abroad on the tissue culture of chrysanthemum are as follows: 1) Propagation by stem section with buds, but the speed of propagation is slow and low efficiency, which is far less than that induced by adventitious buds; 2) Cotyledon and hypoembryon The axis is an explant, and the regeneration system is constructed by induction of adventitious buds, but these materials are too young, easy to be damaged and inconvenient to operate during the establishment of the genetic transformation system, and there is a lack of controls with consistent genetic backgrounds when using this material for transgenic function research. plants, cannot accurately verify gene function

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  • A high-efficiency regeneration method using yellow beam leaves as explants

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Experimental program
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Effect test

Embodiment 1

[0021] (1) Inducing adventitious buds: select the young leaves of the subcultured aseptic seedlings as explants, remove the edges and cut 2-3 cuts horizontally to increase the wound. Inoculated on the induction medium, cultured in total darkness at 25°C for 3 days, then transferred to a light intensity of 1500 lx, light time of 12 hours, and cultured at a temperature of 25°C until adventitious buds were induced, and adventitious bud induction The rate can reach 92.7%. The induction medium is: MS+0.1mg / L TDZ+0.5mg / L 2,4-D+1.0mg / L IBA+20g / L sucrose+4.5g / L agar, the pH is 5.5;

[0022] (2) subculture: the adventitious buds obtained in step (1) are cut off from the base, inoculated on the proliferation medium and carried out subculture, after inoculation, place the light for 13 hours every day, the light intensity is 2500lx, and the culture temperature is 25 Cultivate under the condition of ℃, transfer once every 20 days, and the proliferation coefficient is 7.64. The proliferat...

Embodiment 2

[0026] (1) Inducing adventitious buds: select the young leaves of the subcultured aseptic seedlings as explants, remove the edges and cut 3 times across the main veins to increase the wound. Inoculated on the induction medium, cultured in total darkness at 26°C for 4 days, then transferred to a light intensity of 2000lx, light time of 13 hours, and cultured at 26°C until adventitious buds were induced, and adventitious bud induction The rate can reach 95.4%. The induction medium is: MS+0.2mg / L TDZ+1.0mg / L 2,4-D+2.0mg / L IBA+30g / L sucrose+3.5g / L agar, pH is 5.4;

[0027] (2) subculture: the adventitious buds obtained in step (1) are cut off from the base, inoculated on the proliferation medium and carried out subculture, after inoculation, place the light for 14 hours every day, the light intensity is 2000lx, and the culture temperature is 26 Cultivate under the condition of ℃, transfer once every 25 days, and the proliferation coefficient is 8.13. The proliferation medium is:...

Embodiment 3

[0031] (1) Inducing adventitious buds: select the young leaves of the subcultured aseptic seedlings as explants, remove the edges and cut 3 times across the main veins to increase the wound. Inoculated on the induction medium, cultured in total darkness at 28°C for 4 days, then transferred to a light intensity of 2500lx, light time of 15 hours, and cultured at a culture temperature of 28°C until the formation of adventitious buds was induced, and adventitious bud induction The rate can reach 89.1%. The induction medium is: MS+0.3mg / L TDZ+1.5mg / L 2,4-D+3.0mg / L IBA+15g / L sucrose+6.0g / L agar, pH is 5.4;

[0032] (2) Subculture: the adventitious buds obtained in step (1) are cut off from the base, inoculated on the proliferation medium and carried out subculture, after inoculation, place the light for 15 hours every day, the light intensity is 2500lx, and the culture temperature is 28 Cultivate under the condition of ℃, transfer once every 20 days, and the proliferation coefficie...

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Abstract

The invention discloses a method for efficient regeneration with a neolamarckia cadamba leaf as an explant, and belongs to the technical field of plant tissue culture. Adoption of the leaf as the explant has the great advantages of ready availability of the material, easiness for operation, extensive sources, favorability for repeated regeneration and genetic transformation and the like. Therefore, an aseptic tender seedling leaf of neolamarckia cadamba is adopted as the explant to successfully obtain an in-vitro regenerated plant of the neolamarckia cadamba and establish an efficient regeneration system for the neolamarckia cadamba by processes of adventitious bud induction, proliferation, rooting, acclimatization and transplantation and the like. The method is significant for realizing rapid propagation and large-scale popularization of an excellent neolamarckia cadamba variety and promoting industrial development of the neolamarckia cadamba, and a foundation is laid for construction of a genetic transformation system for the neolamarckia cadamba, so that a platform is provided for molecular biological researches on gene functions and the like of the neolamarckia cadamba.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a high-efficiency regeneration method using yellow beam leaves as explants. Background technique [0002] Neolamarckia cadamba, also known as tuanhua tree, belongs to the evergreen broad-leaved tree of Rubiaceae Tuanhua, widely distributed in South China (Ouyang et al, 2013). The height can reach 35m, the diameter at breast height can reach 1m, and the trunk is straight and round. It takes about 10 years to mature, and the diameter at breast height can reach 40-50cm. The decline period is late, and it is suitable for cultivating large-diameter timber. The high growth period is 5 years ago, and the annual average growth rate is high. The volume reaches 3.0-3.5m, and the diameter growth period is 5-10 years, with an average annual growth volume of 3-4cm (Deng Xiaomei et al., 2011). Because of its rapid growth, it was hailed as the "jewel tree" in the Phili...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/008
Inventor 欧阳昆唏李景剑陈晓阳张俊杰李俊成李培周玮
Owner SOUTH CHINA AGRI UNIV