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Method for establishing in-vitro regeneration system of Acrostichum aureurm

A technology of in vitro regeneration and brine fern, applied in the field of agricultural biology, can solve the technical bottlenecks of seedling production and other problems, and achieve the effect of highly intensive and high-density factory production and reliable technical support

Inactive Publication Date: 2016-01-13
NANCHANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The plants currently used for mangrove restoration are all obtained through seed propagation and viviparous seedling propagation. As the only fern group in true mangrove plants, Halofern ( Acrostichum ) is rarely used, which is related to the bottleneck of the seedling production technology of the halogenated fern. At present, there is no report on the establishment method of the in vitro regeneration system of the halogenated fern.

Method used

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  • Method for establishing in-vitro regeneration system of Acrostichum aureurm
  • Method for establishing in-vitro regeneration system of Acrostichum aureurm
  • Method for establishing in-vitro regeneration system of Acrostichum aureurm

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Explant collection, surface disinfection and inoculation: select fresh sporangia with dark brown sporangia, cut the sporophyll and bring it back in a ziplock bag, and process the fresh material in time. Surface disinfection of sporophylls: cut sporophylls into strip-shaped fragments of 1×3~4cm, put them in a 100ml jar, soak in saturated washing powder solution, add 1 drop of Tween-80, cover the bottle cap, and soak for 20 minutes. Gently swirl the bottle several times every 3~5 minutes; tie the mouth of the wide-mouth bottle with gauze, and wash the spore leaves slowly with tap water for 10 minutes until the liquid in the bottle is clear; on the ultra-clean workbench, discard the tap water in the bottle and add % alcohol soak for 20s, discard the alcohol, add 0.1% HgCl 2 Disinfect for 2.5 minutes, then rinse 5 times with sterile water, and set aside. Use a scalpel to scrape the sporangia group on the ventral surface of the sporophyll, connect it to the surface of the s...

Embodiment 2

[0048] The prothallium obtained in Example 1 was inoculated with the prothallus and cut into the prothallium proliferation medium for prothallium proliferation culture. The culture medium was replaced every 50 days. The culture conditions are: 12h light + 12h dark, light intensity 2500Lx, 25±2°C.

[0049] Prothallus proliferation medium formula is:

[0050] a) 1 / 4MS basic medium + 15g / L sucrose + 6-BA0mg / L + 2,4-D0.1mg / L + agar 7g / L.

[0051] b) 1 / 4MS minimal medium + 15g / L sucrose + 6-BA0.2mg / L + 2,4-D0.1mg / L + agar 7g / L.

[0052] c) 1 / 4 MS basic medium + 15g / L sucrose + 6-BA0.4mg / L + 2,4-D0.1mg / L + agar 7g / L.

[0053] d) 1 / 4 MS basic medium + 15g / L sucrose + 6-BA0.6mg / L + 2,4-D0.1mg / L + agar 7g / L.

[0054] After culturing for 50 days, the results are as follows:

[0055] a) Fragments of prothallium can form new prothallium, but the number is limited. The ventral rhizome of the prothallus is well developed.

[0056] b), c), d) prothalloid fragments can form new prothal...

Embodiment 3

[0059] The prothallus obtained in Example 2 was pulverized and inoculated into the sporophyte induction medium to induce the formation of seedlings. The culture medium was replaced every 50 days. The culture conditions are: 12h light + 12h dark, light intensity 2500Lx, 25±2°C.

[0060] The formula of sporophyte induction medium is:

[0061] a) 1 / 4MS minimal medium + GA 3 0mg / L+5g / L sucrose+agar 7g / L+hydrolyzed casein 100mg / L.

[0062] b) 1 / 4MS minimal medium + GA 3 0.4mg / L+5g / L sucrose+agar 7g / L+hydrolyzed casein 100mg / L.

[0063] c) 1 / 4MS minimal medium + GA 3 0.8mg / L+5g / L sucrose+agar 7g / L+hydrolyzed casein 100mg / L.

[0064] d) 1 / 4MS minimal medium + GA 3 1.2mg / L+5g / L sucrose+agar 7g / L+hydrolyzed casein 100mg / L.

[0065] After culturing for 50 days, the results are as follows:

[0066] a) Sporophyte seedlings are formed, with an average of 20 seedlings / bottle, and the height of young sporophylls is ≤0.6cm. The young leaves are relatively thick, and some veins are v...

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Abstract

The invention discloses a method for establishing an in-vitro regeneration system of Acrostichum aureurm. According to the method, sporangiorus is used as explant; after surface disinfection, the explant is inoculated into a variety of mediums containing a 1 / 4 MS basal medium; prothallus is obtained through in-vitro germination of spores in sporangium; a part of the prothallus can further undergo enrichment culture, while the other part of the prothallus can be used for induced production of juvenile sporophyte; and a sapling is transplanted into a matrix after rooting culture, and the seedling of Acrostichum aureurm is formed eventually. The method can obtain a great number of high-quality seedlings in a short period of time, fills a technical blank in the prior art and provides powerful technical guarantee for garden cultivation, research and development of traditional Chinese medicine and ecological restoration of mangrove forest.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and relates to a method for establishing a halide fern in vitro regeneration system with sporangia group as explants. Background technique [0002] Halogen fern ( Acrostichumauream L.) is a plant of the genus Halogenaceae, which is distributed in tropical and subtropical regions of Asia, Africa, America, and Oceania. It often grows on the inner edge of mangrove plants on tidal flats or on the banks where tidal power reaches. It is an important symbol species of coastal tidal flats. [0003] Halogen fern has high ornamental value. The plant is tall and straight, up to 2m. The rhizome is erect, and the top is densely covered with brown-brown broad-lanceolate scales. The leaves are thick and leathery, clustered; the base is pinnately compound, with up to 30 pairs of pinnae, the petiole is 30-60cm long and 2cm thick; the leaves are 60-140cm long and 30-60cm wide; the veins are reticula...

Claims

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Application Information

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IPC IPC(8): A01H4/00A01G31/00
Inventor 蔡奇英王保忠杨志旺刘以珍葛刚
Owner NANCHANG UNIV
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