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A kind of tissue culture propagation method of golden retriever dog green spheroid pathway

A technology of green spheroids and golden retrievers is applied in the field of plant tissue culture to achieve the effects of inhibiting differentiation, high reproductive efficiency and high proliferation rate

Active Publication Date: 2017-01-04
FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, at present, there is no relevant report on the research on the tissue culture reproduction of the rare and endangered fern Golden Retriever through the green spheroid approach.

Method used

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  • A kind of tissue culture propagation method of golden retriever dog green spheroid pathway
  • A kind of tissue culture propagation method of golden retriever dog green spheroid pathway
  • A kind of tissue culture propagation method of golden retriever dog green spheroid pathway

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Method of the invention

[0041] (1) Obtaining sterile explants of golden retriever dogs

[0042] Inoculate the golden retriever's sterile spores on a medium of 1 / 4MS+sucrose 30.0g / L+agar 7.0g / L, pH 5.80, culture conditions: 20~25℃, daily light time of 12 hours, light intensity 2000lx, get young and sterile seedlings. In the ultra-clean workbench, the obtained sterile seedlings are peeled off the stem tip with tweezers under a dissecting microscope, and the stem tip is used as a sterile explant induced by the golden dog green spheroid.

[0043] The method of obtaining aseptic spores of Golden Retriever is: put 10mg spores in a 1.5ml centrifuge tube, soak in sterile water for 1-2 hours, centrifuge at 6000r / min for 2min to obtain spore precipitate, add 5% NaClO solution in volume fraction Sterilize for 4 minutes and wash with sterile water for 3-5 times to obtain sterile golden retriever spores.

[0044] (2) Induction of green spheroids of golden retriever dog

[0045]...

Embodiment 2

[0062] Example 2 and Example 3 are both methods of the present invention

[0063] In Example 2 and Example 3, except for the different concentrations of the components in each culture medium listed in Table 1, the other measures are the same as in Example 1, and will not be repeated.

[0064] Table 1 The difference between the steps of Example 2 and Example 3 and Example 1

[0065]

[0066]

[0067] The reproduction results of Example 1: The induction rate of green spheroids is 94%, the differentiation rate is 100%, and the rooting rate is 100%, that is, the formation rate of tissue cultured seedlings is 100%. The rooting culture is 28 days, and the average height of the tissue cultured seedlings is 3.15cm . The fresh weight of the green spheroids with a diameter of 2mm is 8mg, the average fresh weight of the green spheroids increases by 76mg every 18 days, and the proliferation multiple is 9.5 times on average. The obtained green spheroids are intermediate propagules, which are su...

Embodiment 3

[0069] The reproduction results of Example 3: The induction rate of green spheroids is 92%, the differentiation rate is 100%, and the rooting rate is 100%, that is, the formation rate of tissue cultured seedlings is 100%. The rooting culture is 28 days, and the average height of the tissue cultured seedlings is 3.30cm . The fresh weight of green spheroids with a diameter of 2mm was 8mg, and the average fresh weight of green spheroids increased by 77.6mg every 21 days, and the average proliferation multiple was 9.7 times. The obtained green spheroids are intermediate propagules, which are subjected to two-step propagation culture for 42 days, differentiation culture for 28 days, rooting culture to form tissue culture seedlings for 28 days, and propagation time for 98 days.

[0070] A 1-3mm green spheroid can obtain 20-25 tissue culture seedlings once differentiated.

[0071] The foregoing Examples 1 to 3 show that the induction rate of the green spheroids of the present invention i...

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Abstract

The invention discloses a tissue culture propagation method for obtaining a cibotium barometz green globular body. The method includes the steps of obtaining an aseptic explant; inducing the green globular body, wherein the green globular body is obtained after being cultivated in two inducing pre-culture media and then cultivated in a green globular body inducing culture medium; cultivating the green globular body in two propagation culture media alternately, and conducting differentiation culture and rooting culture to obtain a tissue culture seedling. By means of the method, the rare and endangered fern type cibotium barometz green globular body is successfully induced, the propagation efficiency is high, the inducing rate of the green globular body reaches 90-94%, the propagation cycle is short, the method has great significance in increasing the population quantity of rare and endangered fern type cibotium barometz and relieving the endangered situation, and meanwhile the in-vitro conservation of germplasm resources of endangered species is achieved.

Description

Technical field [0001] The invention belongs to the technical field of plant tissue culture, and specifically relates to a method for inducing and multiplying golden retriever green spheroids. [0002] technical background [0003] Golden Retriever (Cibotium barometz), belonging to the genus Golden Retriever of the Clamshell Fern family, has a prone, thick rhizome, and an upturned end. The exposed part of the ground is densely covered with golden yellow hairs, resembling a golden retriever dog with a unique shape. With ornamental value, the golden hair can be used as medicine, and it is included in the "Pharmacy of the People's Republic of China". The "List of National Key Protected Wild Plants (First Batch)" lists all species of Pteridaceae as national-level II key protected wild plants. There is only 1 genus and 1 species of Pteridaceae in mainland China, namely: Golden Retriever Golden retriever dog. [0004] Under natural conditions, golden retrievers use spores for reproductio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 余蓉培王继华莫锡君蒋亚莲田敏李进昆
Owner FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI