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Virus-elimination method of plant tissue cultured virus-free explant

A technology for plant tissue culture and explants, applied in the field of detoxification of plant tissue culture detoxification explants, can solve the problems of explant detoxification culture failure, etc., achieve simple operation of tissue culture detoxification, increase differentiation rate and The effect of high survival rate, differentiation rate and survival rate

Inactive Publication Date: 2017-05-17
DALIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In order to solve the problem that the explants used in the shoot tip detoxification culture method in the prior art are too small to cause the failure of detoxification culture, the present invention adopts mature tissue [1] As an explant, it makes the tissue culture detoxification operation easier, greatly improves the differentiation rate and survival rate, and makes the detoxification success rate higher

Method used

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  • Virus-elimination method of plant tissue cultured virus-free explant
  • Virus-elimination method of plant tissue cultured virus-free explant

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Choose the leaves of the dicotyledonous plant "Red Fuji" apple in the growth period as explants, wash them in running water, sterilize them in 30% garlic aqueous solution for 30 minutes, and then cut them into 1cm 2 The squares were immediately inoculated on the optimal primary medium (MS+2,4-D 1.0mg / L+3% sucrose+agar 8g / L). After the callus is induced, it is transferred to the most suitable subculture medium (2 / 3MS+6-BA0.6mg / L+IBA0.2mg / L+GA1.0+3% sucrose+8g / L agar) for cultivation, Proliferation and cultivation are carried out continuously, and when the subcultured rootless seedlings reach a large number, 50-100 mg of leaves are taken for detection of main viruses. The virus detected this time is one of the main viruses in apples, that is, apple chlorotic leaf spot virus (ACLSV), such as figure 1 There is no virus-specific fragment with a length of 358bp in the electrophoresis graph shown, that is, the test result is negative, and the subculture seedlings with a negat...

Embodiment 2

[0030] The phloem of the dicotyledonous plant "Red Fuji" apple in the dormant period was selected as explants, washed with running water, sterilized in 50% garlic aqueous solution for 30 minutes, and immediately inoculated in the optimal primary medium (MS+2,4-D1 .5mg / L+3% sucrose+agar 8g / L). After the callus is induced, it is transferred to the most suitable subculture medium (2 / 3MS+6-BA 0.6mg / L+IBA0.2mg / L+GA1.0+3% sucrose+8g / L agar) for cultivation, Proliferate and cultivate continuously, and when the subcultured rootless seedlings reach a large number, take 50-100 mg of leaves to detect the main virus ACLSV. The test result is then negative (i.e. electrophoresis figure 1 The progeny seedlings that do not have a length of 358bp virus-specific fragment) continue to be cultivated in the proliferation medium, and after reaching a certain number, they are transferred to a suitable rooting medium (1 / 2MS+IBA1.0mg / L+3 % sucrose+8g / L agar) to carry out rooting culture, transfer to...

Embodiment 3

[0032] Choose the leaves of the monocotyledon Phalaenopsis, rinse with running water, soak in 30% garlic solution for 30min, and then cut into 1cm 2 Immediately inoculated on the optimal primary medium (MS+2,4-D1.0mg / L+BA1.0mg / L+3% sucrose+agar 8g / L). After the callus is induced, it is transferred to the most suitable subculture medium (MS+6-BA 1.0mg / L+IBA0.2mg / L+3% sucrose+8g / L agar) for cultivation, and the proliferation culture is continuously carried out. When the subcultured rootless seedlings reach a large number, 50-100 mg of leaves are taken to detect the main virus. The virus detected this time is one of the main viruses of Phalaenopsis, namely Cymbidium mosaic virus (CyMV). The test result is then negative (i.e. electrophoresis figure 2 The progeny seedlings that do not have a length of 431bp virus-specific fragment) continue to be cultivated in the proliferation medium, and after reaching a certain number, they are transferred to a suitable rooting medium (1 / 2MS+...

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Abstract

The invention belongs to the technical field of virus elimination of plants, and particularly relates to a virus-elimination method of a plant tissue cultured virus-free explant. The method comprises the following steps: selecting plant mature tissues, performing sterilizing treatment on the selected plant mature tissues, performing primary culture on the sterilized tissues in a primary culture medium, performing propagation culture based on the primary culture, performing virus detection based on the propagation culture, sorting secondary seedlings of which the viruses are negative, extending reproductive output, then performing rooting culture until favorable root systems grow, and then performing domestication and transplanting. Mature tissues are used as explants, so that the tissue culture virus elimination operation is simple, the differentiation rate and the survival rate are greatly increased, and the virus-elimination success rate is high.

Description

technical field [0001] The invention belongs to the technical field of plant detoxification, in particular to a detoxification method for plant tissue culture detoxification explants. Background technique [0002] Viruses are seriously harmful to plants. Viruses can lead to weakening of plant trees, smaller and deformed fruits, poor quality, and serious decline in yield. Virus diseases are difficult to treat, and so far there is no effective drug to solve them. [2-7] . Therefore, cultivating virus-free plant seedlings has become the main measure to prevent virus damage. At present, the method usually adopted to cultivate virus-free plant seedlings is shoot tip tissue culture technology. Shoot tip culture can remove virus from plant tissue, resulting in virus-free (or virus-free) plants. Shoot tip culture, sometimes called micro shoot tip culture, utilizes the principle that the shoot tip is a meristematic tissue, and there is no virus in it, and the purpose of virus remov...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/008
Inventor 侯义龙
Owner DALIAN UNIV
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