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Method for sterile seeding of lithops and establishing regeneration system

A technique for aseptic sowing and lithotripsy, which is applied in the field of plant tissue culture technology and plant shoot tip culture, and can solve problems such as unsuccessful examples.

Active Publication Date: 2015-06-03
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, before this, there is no report on the establishment of Lithops tissue culture regeneration system, let alone successful examples

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The present invention provides a kind of Lithops aseptic sowing and regeneration system establishment method, and its steps are:

[0027] 1) Preparation of culture medium

[0028] (1) Basic medium: MS medium, in which sucrose 30g / L, agar 7.5g / L, pH=5.8;

[0029] (2) Induction medium: MS+6-BA0.1~0.5mg / L+NAA0.05~0.1mg / L;

[0030] (3) Differentiation medium: MS+6-BA0~0.05mg / L;

[0031] (4) Proliferation medium: MS+6-BA0.05~0.3mg / L+NAA0.05~0.1mg / L;

[0032] 2) Disinfection and inoculation of seeds

[0033] Take Lithops seeds as explant material, put them in 1.5ml or 2ml PE centrifuge tubes for disinfection treatment, first soak them in detergent solution for 0.5h, then wash them with tap water, and then wash them in a volume ratio of 70 % alcohol solution and sodium hypochlorite solution with an effective chlorine concentration of 1% for 30s and 6min, respectively, and finally rinsed with sterile water for 3 to 5 times, each time for 2min (operated with a pipette).

[...

Embodiment 2

[0041] The present invention provides a kind of Lithops aseptic sowing and regeneration system establishment method, and its steps are:

[0042] 1) Preparation of culture medium

[0043] (1) Basic medium: MS medium, in which sucrose is 20-30g / L, agar is 5-9g / L, pH=5.8;

[0044] (2) Induction medium: MS+6-BA0.2mg / L+NAA0.05mg / L;

[0045] (3) Differentiation medium: MS+6-BA0.05mg / L;

[0046] (4) Proliferation medium: MS+6-BA0.1mg / L+NAA0.05mg / L;

[0047] 2) Disinfection and inoculation of seeds

[0048] Take Lithops seeds as explant material, put them in 1.5ml or 2ml PE centrifuge tubes for disinfection treatment, first soak them in detergent solution for 0.5h, then wash them with tap water, and then wash them in a volume ratio of 70 % alcohol solution and sodium hypochlorite solution with an effective chlorine concentration of 1% for 30s and 6min, respectively, and finally rinsed with sterile water for 3 to 5 times, each time for 2min (operated with a pipette).

[0049] 3) I...

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PUM

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Abstract

The invention discloses a method for sterile seeding of lithops and establishing a regeneration system. The method specifically comprises the following operation steps: (1) medium preparation, specifically comprising preparation of minimal medium and mediums at various culturing stages; (2) seed disinfection and inoculation; (3) induction and differentiation of a callus; (4) proliferation of adventitious buds; (5) strong seedling culturing. The method has the beneficial effects that the plant tissue culture technology is adopted for carrying out sterile seeding and generated rapid propagation on the lithops, so that a large number of high-quality strong seedlings with consistent inheritable characters are obtained within short periods, the defect of low speed of the conventional propagation method is overcome, and the method has positive significances for large-scale production of seedlings of lithops and collection and storage of germplasm resources.

Description

technical field [0001] The invention relates to plant tissue culture technology and plant shoot tip culture technology, and mainly relates to a method for aseptic seeding and regeneration system establishment of lithopsia. Background technique [0002] Lithops pseudoruncatella (Lithops pseudoruncatella) is a plant of the genus Lithops in the Apricot family. Because of its unique shape and colorful colors, it is a very popular small ornamental succulent. Its natural reproduction rate is low, and its origin is not in my country. In addition, due to excessive human excavation and destruction of its habitat, the current quantity is small and the market price remains high, which limits its promotion. Therefore, the rapid propagation of Afang Gongshou by plant tissue culture technology is of great significance for the preservation of excellent germplasm resources and the propagation of famous and rare varieties, so as to realize its large-scale production to meet the needs of dom...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 牟豪杰王燕陈剑平汪一婷吕永平陈志
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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