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Vitro quick di-wu breeding method

A kind of Diwu, rapid technology, applied in the field of plant tissue culture, can solve the problems such as the depletion of Diwu wild resources, low vitality of Diwu seeds, and the rapid propagation of Diwu has not been reported, and achieve good proliferation effect and somatic embryo differentiation rate Significant, significant effect of pseudocorm induction rate

Active Publication Date: 2009-03-25
广州康和药业有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the rarity of wild Diwu, the development of Chinese patent medicines for Diwu cannot rely solely on wild resources, and must be cultivated on a large scale. Otherwise, the wild resources of Diwu will be exhausted and the natural ecological environment will be destroyed.
[0004] Through inspection, the vigor of the seeds is very low. At present, it is difficult to directly germinate the seeds to produce the seedlings of the seeds in the laboratory.
Therefore, the seedlings of large-scale artificial cultivation of Diwu at this stage are only obtained through the in vitro propagation (tissue culture) of Diwu, and there is no report about the in vitro rapid propagation of Diwu in domestic and foreign literature.

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0034] Embodiment 1 (general culture program)

[0035] A method for in vitro rapid propagation of ground crow, the steps are as follows:

[0036] (1) Selection and disinfection of explants:

[0037] 1) select the dormant buds with the length of 1-2cm of the old tissue of the Rhizome of Rhizome and the unexpanded leaves of the just germinated as explants;

[0038] 2) The explant is sterilized by the following method:

[0039] a. Disinfection of explants as dormant buds: first wash off the silt on the surface of the buds of the root of the root of the root of the root of the root, and use a soft brush to clean the dead tissue on the surface of the shoot of the root of the root of the root until the bud appears white. Soak in 1.5% carbendazim aqueous solution for 2 hours, rinse with tap water for 0.5 hours, transfer to ultra-clean workbench, disinfect with 75% alcohol for 30-60 seconds, rinse with sterile water for 1-2 times, and then use 0.2% mercuric chloride solution Disinf...

Embodiment 2

[0074] Test material selection, medium design and inoculation culture

[0075] 1. Sources and processing of experimental materials

[0076] The rhizomes of wild black root from Machi Tonggubao, Changyang Tujia Autonomous County, Hubei Province, at an altitude of about 1,800 meters were selected as test materials. Before use, use the method introduced by the applicant's prior invention patent application (patent application number: 200810047075.X, publication number: CN101243741, title of invention: the method for removing the dormancy of the rhizome of the Radix Rhizome) of the applicant. It germinates, grows, and provides explants for testing. The selection criteria and induction results of Diwu explants are shown in Table 1:

[0077] Table 1 Selection and induction results of suitable explants for in vitro propagation of Diwu

[0078]

[0079] Table 1 shows that the Radix Radix bud body is the best explant of the present embodiment, followed by the unexpanded Radix Rad...

Embodiment 3

[0095] Effects of Different Concentrations of Naphthaleneacetic Acid (NAA) on In vitro Propagation

[0096] In plant tissue culture, 6-BA is a widely used auxin growth regulator. It is recognized as an effective auxin for initiating cell dedifferentiation to form callus or organogenesis.

[0097] After cultured for 2 weeks, white and transparent callus tissue can be seen from the incision. After subculture once in 30 days, and after 2 subcultures, three different regeneration tissues can be seen, namely, adventitious buds, pseudocorms and Somatic embryos correspond to three different regeneration pathways, adventitious bud regeneration pathway, pseudocorm regeneration pathway and somatic embryogenesis pathway. It was observed that when the concentration of 6-BA was 2.0mg / L, the concentration change of NAA would lead to the occurrence of different regeneration pathways,

[0098] Table 3 Effects of different concentrations of NAA on Diwu propagated in vitro

[0099]

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Abstract

The invention belongs to the technical field of plant tissue culture, particularly relating to a rapid in-vitro propagation method for rhizome of flaccid anemone; the method comprises explant selection and sterilization of the rhizome of flaccid anemone, inducement and proliferation of the adventitious buds, pseudobulb inducement and proliferation, embryogenic callus inducement, proliferation, differentiation, rooting and other steps. The invention provides an explant aseptic sterilization and cultivation system for rhizome of flaccid anemone, a cultivation process, a special culture medium and other key techniques, and successfully produces the tube-cultivated plantlet of the rhizome of flaccid anemone. The invention overcomes the disadvantages of long breeding cycle and low breeding coefficient of the rhizome of flaccid anemone under natural conditions, and provides a practical technology for rapidly propagating tube-cultivated plantlets of the rhizome of flaccid anemone in an industrialized mode.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for in vitro rapid propagation of Aconiti japonica. technical background [0002] Diwu, whose scientific name is Anemone Flaccida Fr. Schmidt, is a perennial herb belonging to the Ranunculaceae Anemone plant, and its dry rhizome is called Diwu, commonly known as Diwu. From March to May every year is the growth and development period of the aboveground part of Diwu. Basal leaves 1-2, long petiole, pentagonal, 2-7cm long, 3-10cm wide. The rhizome is nearly cylindrical, with a maximum diameter of 1 cm, and it can be harvested as medicine in 3 to 5 years. The wild resources of Diwu are mainly distributed in the damp places under the hillside forest at an altitude of 1500-3000m in the middle and lower reaches of the Yangtze River, and the distribution in the mountains of western Hubei is relatively concentrated. From this point of view, the distrib...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00C12N5/04
Inventor 朱端卫张忠池杨特武耿明建黄漫翔方贤东李宏飞王俊辛龙
Owner 广州康和药业有限公司
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