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A method for establishing a high-efficiency in vitro regeneration system of Qishu mature embryos

A technology for in vitro regeneration and mature embryos, applied in the field of plant biology, can solve the problems of low reproduction coefficient, low seed germination rate, and ecological environment damage, etc., and achieve the effect of increasing reproduction coefficient, growth ability and regeneration rate

Active Publication Date: 2022-04-19
HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, due to the slow growth of Qishu, less flowering and fruiting, some seeds are not viable, and with the increasing amount of folk excavation, the ecological environment of Qishu is severely damaged, resulting in the distribution of wild Qishu in the natural ecological environment. rare
At present, in terms of production, Qishu usually adopts the traditional sowing method for propagation, but there are problems such as low seed germination rate, weak seedling resistance, and vulnerable to various diseases and insect pests such as blight and root rot.
In addition, it is also feasible to use rhizomes to propagate Qishu. However, since rhizomes are the main medicinal parts of Qishu, the cost of propagation in this way is high, the propagation speed is slow, the propagation is limited by materials, and it is easy to introduce diseases, etc. question
At present, there have been reports on the propagation of Qishu by using tissue culture technology, but there are problems such as incomplete disinfection and low reproduction coefficient in the reproduction process, which seriously limits the large-scale production and popularization of Qishu seedlings.

Method used

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  • A method for establishing a high-efficiency in vitro regeneration system of Qishu mature embryos
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  • A method for establishing a high-efficiency in vitro regeneration system of Qishu mature embryos

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Embodiment 1

[0033] A method for establishing a high-efficiency in vitro regeneration system of Qishu mature embryos, the specific operations are as follows:

[0034] (1) Select vigorous and mature seeds of Qizhu as explants, place them in a 250ml plastic culture bottle, and rinse them under running water for 10 minutes. Then put the seeds in the aseptic operating table, rinse them with sterile water for 3 times, and then use 75% absolute ethanol to carry out surface disinfection twice, each time for 15 seconds, rinse them twice with sterile water between the two times, and then use 0.1% (w / v) mercuric chloride solution was sterilized for 3 minutes, and then rinsed 6 times with sterile water; further above-mentioned seeds were filtered and sterilized with low-temperature plasma-activated water (Ar was the ion source, and treated with a dielectric barrier discharge device). steamed water for 2 minutes) soaked for 2 minutes, then dry the moisture on the surface of the seeds with sterile filt...

Embodiment 2

[0040] A method for establishing a high-efficiency in vitro regeneration system of Qishu mature embryos, the specific operations are as follows:

[0041] (1) Select vigorous and mature seeds of Qizhu as explants, place them in a 250ml plastic culture bottle, and rinse them under running water for 20 minutes. Then put the seeds in the aseptic operating table, rinse them with sterile water for 4 times, and then use 75% absolute ethanol to carry out surface disinfection for 2 times. 0.1% (w / v) mercuric chloride solution was sterilized for 4 minutes, and then rinsed 8 times with sterile water; then the above seeds were filtered and sterilized by low-temperature plasma-activated water (Ar was the ion source, and treated with a dielectric barrier discharge device). steamed water for 2 minutes) soaked for 3 minutes, and finally dry the moisture on the seed surface with sterile filter paper, and set aside.

[0042] (2) The pretreated seeds in step (1) were inoculated in the induction...

Embodiment 3

[0047] A method for establishing a high-efficiency in vitro regeneration system of Qishu mature embryos, the specific operations are as follows:

[0048] (1) Select vigorous and mature seeds of Qizhu as explants, place them in a 250ml plastic culture bottle, and rinse them under running water for 15 minutes. Then put the seeds in the aseptic operating table, rinse them with sterile water for 3 times, and then use 75% absolute ethanol to carry out surface disinfection twice, each time for 17 seconds, rinse them twice with sterile water between the two times, and then use 0.1% (w / v) mercuric chloride solution was sterilized for 5 minutes, and then rinsed 7 times with sterile water; further above-mentioned seeds were filtered and sterilized by low-temperature plasma activated water (Ar was the ion source, and treated with a dielectric barrier discharge device). steamed water for 2 min) and soaked for 4 min, then use sterile filter paper to dry the water on the surface of the seed...

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Abstract

The invention discloses a method for establishing a high-efficiency in vitro regeneration system for mature embryos of Qizhu, comprising the following steps: selecting and pretreating seeds, applying low-temperature plasma activated water to the pretreatment process; pretreating mature embryos, Inoculate in the adventitious bud induction medium for the induction culture of adventitious buds; transfer the mature embryos that induce adventitious buds to the proliferation medium for the proliferation and culture of adventitious buds; cut the proliferated adventitious bud clusters into cut pieces of appropriate size Then transfer them to the elongation medium for elongation culture of adventitious buds; when the adventitious buds grow to 2-3 cm with complete leaves, transfer them to rooting medium for rooting culture, and finally obtain complete regeneration plants. The present invention applies low-temperature plasma activated water to the pretreatment process, and then realizes the rapid propagation of qizhu seedlings by directly inducing the regeneration of clustered buds, and provides a powerful technology for the efficient breeding and large-scale production of qizhu seedlings ensure.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, and in particular relates to a method for establishing a high-efficiency in vitro regeneration system of mature embryos. Background technique [0002] Qishu is a medicinal perennial plant of the Atractylodes family of Asteraceae. It is named after it is produced in Qimen. Its rhizome can be used as medicine. It is one of the most representative Chinese medicinal materials in Qimen, and it is also a rare and precious medicinal material in Anhui. A large number of clinical studies of traditional Chinese medicine have shown that Qizhu is warm in nature, sweet in taste, and slightly bitter. In addition to the general effects of Atractylodes macrocephala invigorating the spleen and replenishing qi, drying dampness and diuresis, nourishing the mind, preventing miscarriage, and eliminating fatigue, it is also effective for jaundice, heart disease, and gastric ulcer. And ascites disease has a significa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/008A01H4/001
Inventor 侯金艳吴丽芳
Owner HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI