Green globular body tissue culture propagation method for Alsophila costularis

A technology of green spheroids and cypress, which is applied in the field of plant tissue culture to achieve the effects of high proliferation rate and high reproductive efficiency

Active Publication Date: 2015-12-16
FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, at present, there is no relevant report on the research on the tissue culture and pr...

Method used

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  • Green globular body tissue culture propagation method for Alsophila costularis
  • Green globular body tissue culture propagation method for Alsophila costularis
  • Green globular body tissue culture propagation method for Alsophila costularis

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Experimental program
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Embodiment 1

[0040] Embodiment 1 The inventive method

[0041] (1) Acquisition of sterile explants of C. sinensis

[0042] Inoculate the sterile spores of Cymbidium sinensis on the medium of 1 / 4MS + 30.0g / L sucrose + 7.0g / L agar, pH 5.80, the culture conditions are: 20~25℃, the light time is 12 hours per day, and the light intensity is 2000lx, to obtain young tender aseptic seedlings. In the ultra-clean workbench, the obtained sterile seedlings were peeled off the shoot tips with tweezers under a dissecting microscope, and the shoot tips were used as sterile explants induced by the green spheroids of C. sinensis.

[0043] The method for obtaining aseptic spores of Cylathelus sinensis is as follows: put 10 mg of spores in a 1.5ml centrifuge tube, soak in sterile water for 1 to 2 hours, centrifuge at 6000r / min for 2 minutes to obtain spore precipitates, and add 5% NaClO solution Sterilize for 4 minutes, and wash with sterile water for 3-5 times to obtain sterile spores of C. sinens...

Embodiment 2

[0062] Embodiment 2 and embodiment 3 are the method of the present invention

[0063] Example 2 and Example 3 except that the concentration of each component in each culture medium listed in Table 1 is different, other measures are the same as Example 1, and will not be repeated.

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

[0065]

[0066]

[0067] The propagation result of embodiment 1: the induction rate of green spheroid is 88%, the differentiation rate is 100%, and the rooting rate is 100%, that is, the formation rate of tissue cultured shoots is 100%, rooted and cultivated for 28 days, and the average height of tissue cultured shoots is 2.87cm . The green spheroid with a diameter of 2mm has a fresh weight of 10 mg, and the average fresh weight of the green spheroid increases by 75 mg every 21 days, and the multiplication factor is about 7.5 times. The obtained green spheroids are intermediate propagules, which a...

Embodiment 3

[0069] The propagation result of embodiment 3: the induction rate of green spheroid is 82%, and the differentiation rate is 100%, and the rooting rate is 100%, that is, the formation rate of tissue cultured shoots is 100%, rooted and cultivated for 28 days, and the average height of tissue cultured plants is 2.70cm . The green spheroid with a diameter of 2mm has a fresh weight of 10 mg, and the average fresh weight of the green spheroid increases by 83.67 mg every 21 days, and the multiplication factor is about 8.4 times. The obtained green spheroid is an intermediate propagule, which undergoes two-step multiplication 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 1-3 mm green spheroid can be differentiated once to obtain 20-25 tissue cultured seedlings.

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Abstract

The invention discloses a green globular body tissue culture propagation method for Alsophila costularis. The method includes the steps of obtaining aseptic explants and inducing green globular bodies. In the inducing process, culture is performed through two inducing pre-culture media, then culture is performed through a green globular body inducing culture medium, and the green globular bodies are obtained; then, alternate culture is performed through two proliferation culture media, differentiation culture and rooting culture are performed, and tissue culture seedlings are obtained. Through the method, the dendritic woody fern green globular bodies are successfully induced, the propagation efficiency is high, the green globular body inducing rate reaches 82%-88%, the proliferation multiple reaches 7.5-9.3, the differentiation rate of the green globoids reaches 100%, the rooting rate reaches 100%, and the propagation period is short; the method has great significance in increasing the population quantity of the rare endangered fern Alsophila costularis and relieving the endangered situation of the rare endangered fern Alsophila costularis, and meanwhile in-vitro preservation of germplasm resources of endangered species is achieved.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for inducing and proliferating green spheroids of Cyaphila sinensis. technical background [0002] Alsophila costularis (Alsophila costularis) belongs to the family Alsophilaceae (Alsophila genus). The stems above the ground are erect, tree-like, up to 10m high, and are one of the few woody ferns. [0003] Alsophilaceae plants first appeared in the Early Jurassic or Late Triassic of the Mesozoic Era, and are the contemporary species of dinosaurs. Later, due to geological changes and climate changes, a large number of species became extinct, and finally only existed in certain "refuges" with particularly suitable climates in the tropics and subtropics, so they were called "relict plants" and "living fossils" in the plant kingdom. Chinese spinosa and all species of Cylatheaceae are listed as the second-class national key protected wild plants "Li...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 余蓉培李绅崇桂敏蒋亚莲周旭红卢珍红施自明
Owner FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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