Green globular body (GGB) pathway drynaria roosii seedling tissue culture rapid propagation method

A green spheroid, tissue culture rapid propagation technology, applied in horticultural methods, botanical equipment and methods, horticulture and other directions, can solve problems such as unreported, time-consuming operation, low survival rate, etc. Cost, improvement of proliferation efficiency reduction effect

Active Publication Date: 2020-07-14
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some progress has been made in the artificial breeding of Quercus fern. The patent "A Breeding Method of Quercus fern" (Shi Lei and Xu Yan, 2008) reported that sterilized Quercus fern spores were sown on the culture medium to obtain aseptic seedlings. method, which can quickly obtain the sporozoites of Quercus fern, but it is limited by the vigor of the spores. This method still needs to collect the fertile sporophytes of Quercus fern, and properly preserve the collected spores.
Establishing the GGB breeding route of Quercus fern seedlings can break through this limitation, but this method has not been reported so far
In addition, the tissue-cultured seedlings of Quercus fern need to be transplanted out of the bottle when the rhizome is about 0.6-0.8 cm long. Too small rhizome will result in time-consuming operation and low survival rate. So far, there has been no improvement in the promotion of the rhizome of the young sporophyte of Quercus fern in the bottle. fast growing technology

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] (1) Disinfection of explants: select the vigorously grown Quercus fern rhizomes as explants, soak the rhizomes in a detergent solution with a concentration of 5g / L for 8min, rinse them under running water for 60min, and remove them. Blot the surface moisture with filter paper, soak in 75% alcohol solution for 30s, then quickly put it into 0.1% mercuric solution for surface disinfection for 6 minutes, and finally rinse it with sterile water for 5 times.

[0026] (2) Induction of GGB: use 1 / 2MS+2.0mg / L 6-BA+3% sucrose+0.7% agar as the induction medium, cut the rhizome after step (1) into 0.5cm long pieces , inoculated into the induction medium, cultured in the culture room, a small amount of GGB with a diameter of 0.1-0.2 cm can be seen after 55 days, and the GGB induction rate is 35%.

[0027] (3) Proliferation culture of GGB: transfer the GGB obtained in step (2) to the GGB proliferation medium, place it in a culture room for dark cultivation, and the light-proof condit...

Embodiment 2

[0031] (1) With embodiment 1 step (1).

[0032] (2) With embodiment 1 step (2).

[0033] (3) Proliferation culture of GGB: transfer the GGB obtained in step (2) to the GGB proliferation medium, place it in a culture room for dark culture, and the dark condition is to cover double-layer black cloth on the petri dish, and proliferate. The base is 1 / 2MS+2.0mg / L 6-BA+3% sucrose+0.7% agar. After 30 days, the average fresh weight of GGB was 51.6 mg, and the differentiation rate was about 27%; after 60 days, the average fresh weight of GGB was 88.4 mg, and the differentiation rate was about 32%.

[0034] (4) Differentiation culture of GGB and division of bud clusters: inoculate the GGB proliferated in step (3) in the differentiation medium and place it in the culture room for cultivation. The differentiation medium is 1 / 2MS+3% sucrose+0.7% agar . After 30 days, GGB differentiated to produce a large number of clustered buds. At this time, the clustered buds were of different sizes....

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Abstract

The invention provides a green globular body (GGB) pathway drynaria roosii seedling tissue culture rapid propagation method. The method is characterized by comprising the following steps of disinfection of explant; induction of GGB; proliferation culture of GGB; differentiation culture of GGB; bud cluster segmentation; and juvenile sporophyte culture. After the drynaria roosii GGB is obtained, inthe proliferation culture of GGB, the use of cytokinin is reduced, and a dark condition is adopted, so that the reduction of the proliferation efficiency of GGB due to differentiation under illumination conditions is obviously improved, the proliferation speed of GGB is increased, and the use cost of light energy is reduced. After the drynaria roosii GGB is differentiated to generate the bud clusters, the root-shaped stem of the tender juvenile sporophyte obtained by segmenting the bud clusters is extremely small, and the time required for culturing the root-shaped stem to reach the volume which can come out of a bottle can be relatively long. By adding the relatively high-concentration of sucrose to treat the juvenile sporophyte in the culture medium, the growth speed of the late root-shaped stem is greatly increased, and then the propagation period of the seedlings is shortened.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for obtaining tissue culture seedlings of Quercus fern through the green spheroid approach. Background technique [0002] Green Globular Body (GGB) is a special structure that appears in the fern tissue culture system. It is usually a spherical body composed of many green particles, hence the name. GGB can proliferate continuously in the induction medium, and changing the composition of the medium can make it rapidly differentiate into seedlings, which has the characteristics of high proliferation rate, fast differentiation, and short seedling cycle. Therefore, the use of GGB for the propagation of fern seedlings has great development prospects. At present, GGB tissue culture rapid propagation pathways have been established for a variety of ferns, such as staghorn fern (Ye Xiuxian et al., 2020), pen container tree (Zhang Yan et al., 2020), South...

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 石雷王頔李东
Owner INST OF BOTANY CHINESE ACAD OF SCI
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