Method for establishing in-vitro regeneration system of Osmunda vachellii

An in vitro regeneration and systematic technology, applied in the field of agricultural biology, can solve the problems of difficult rapid large-scale cultivation and low reproduction efficiency, 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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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The current resources of Osmanthus chinensis mainly rely on wild resources, but the reproductive efficiency is low under natural

Method used

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  • Method for establishing in-vitro regeneration system of Osmunda vachellii
  • Method for establishing in-vitro regeneration system of Osmunda vachellii
  • Method for establishing in-vitro regeneration system of Osmunda vachellii

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Explant collection and surface disinfection treatment: when the sporangia of Osmanthus chinensis are yellow-brown in color and the sporophyll axis is still yellow-green and fresh, cut the sporophyll and bring it back in a ziplock bag, and process the fresh material in time, or store it at 4°C Save and finish processing within 2d. Surface disinfection of sporophylls: cut sporophylls into small sections of 3~4cm, containing 10-12 sporangia groups, put them in a 50ml jar, soak in saturated washing powder solution, add 1 drop of Tween-80, cap the bottle, soak Wash 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 sporophyll with tap water for 10 minutes until the liquid in the bottle is clear; Tap water, add 70% alcohol to soak for 15s, discard alcohol, add 0.1% HgCl 2 Disinfect for 2 minutes, rinse with sterile water 5 times, and set aside. The sporangia on the sporophylls were separat...

Embodiment 2

[0047] The prothallus obtained in Example 1 was inoculated with the prothallus and cut into the proliferation medium for prothallium proliferation culture. The culture medium was replaced every 50 days. The culture conditions are all: 12h light + 12h dark, light intensity 2500Lx, 25±2°C.

[0048] Proliferation medium formula is:

[0049] a) Improved Beneck basic medium + 15g / L sucrose + 6-BA0mg / L + 2,4-D0.1mg / L + agar 7g / L.

[0050] b) Improved Beneck basic medium+15g / L sucrose+6-BA0.2mg / L+2,4-D0.1mg / L+agar 7g / L.

[0051] c) Improved Beneck basic medium+15g / L sucrose+6-BA0.4mg / L+2,4-D0.1mg / L+agar 7g / L.

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

[0053] After culturing for 50 days, the results are as follows:

[0054] a) Prothallium fragments can form new prothallus, but the number is limited, 22 per bottle on average. The ventral rhizome of the prothallus is well developed.

[0055] b) Prothallium fragments can fo...

Embodiment 3

[0060] 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 all: 12h light + 12h dark, light intensity 2500Lx, 25±2°C.

[0061] The formulation of sporophyte induction medium is:

[0062] a) Modified Beneck basic medium + GA 3 0mg / L+5g / L sucrose+agar 7g / L+hydrolyzed casein 100mg / L.

[0063] b) Improved Beneck basic medium + GA 3 0.4mg / L+5g / L sucrose+agar 7g / L+hydrolyzed casein 100mg / L.

[0064] c) Improved Beneck basic medium + GA 3 0.8mg / L+5g / L sucrose+agar 7g / L+hydrolyzed casein 100mg / L.

[0065] d) Improved Beneck basic medium + GA 3 1.2mg / L+5g / L sucrose+agar 7g / L+hydrolyzed casein 100mg / L.

[0066] After culturing for 50 days, the results are as follows:

[0067] a) Sporophyte seedlings are formed, with an average of 15 seedlings / bottle, and the height of young sporophytes is ≤1cm. The young leaves are ...

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Abstract

The invention discloses a method for establishing an in-vitro regeneration system of Osmunda vachellii, which belongs to the field of agricultural biotechnology. According to the method, sporangiorus is used as explant; after surface disinfection, the explant is inoculated into a variety of mediums containing an improved Beneck 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 (a sapling); and the sapling is transplanted into a matrix after rooting culture, and the seedling of Osmunda vachellii is formed eventually. The method provided by the invention can realize rapid and continuous obtainment of a great number of high-quality seedlings.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and relates to a method for establishing an in vitro regeneration system of Osmanthus chinensis with sporangia group as explants. Background technique [0002] South China purple Osmanthus ( Osmundavachellii Hook.) is a plant of the Osmunda genus in the Osmanthus fern family, which is produced in the humid subtropical region of Asia. Distributed in my country's Hainan Island, Guangdong, Guangxi, Fujian, Jiangxi, Zhejiang, Guizhou, southern Yunnan and Hong Kong, and also produced in Vietnam, Myanmar and India. Often wild in moist mountains, grass or streams, it is an indicator plant for acidic soil in southern my country. [0003] Osmanthus chinensis has high ornamental value. The height of the plant is 1m, the rhizome is erect and cylindrical, and the leaves are clustered on the top, resembling cycads. The plant shape is beautiful and the leaves are elegant. It can be planted in gar...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 蔡奇英杨志旺王保忠刘以珍葛刚
Owner NANCHANG UNIV
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