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Tissue culture bulb preserving method for narcissus pseudonarcissus virus

A preservation method and technology of daffodils, which are applied in the directions of viruses/phages, biochemical equipment and methods, and the determination/inspection of microorganisms, can solve the problems of inconvenient portability and utilization of poison sources, labor-intensive, unsatisfactory effects, etc., to prevent The effect of virus mixing and loss, improving the preservation effect, and being easy to carry and communicate

Inactive Publication Date: 2008-07-09
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional preservation is to inoculate the virus on indicator plants or host plants for regular transfer in the greenhouse, which is a heavy workload and the effect is not ideal
Because a variety of virus source plants are stored in the same greenhouse under bare conditions, it is easy to cause confusion; at the same time, a certain greenhouse area is required, and suitable temperature and humidity conditions are required, especially for heating in winter in cold regions, which consumes manpower and financial resources; The poisonous plants are in the shed, which is easy to be infected with bacteria and die, resulting in the loss of the virus; at the same time, the poisonous source preserved by this method is also inconvenient to carry and use

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] 1) Identification of the pathogenic species of Narcissus virus

[0062] (1) Take disease samples: take the disease samples of daffodil mosaic and leaf disease, and store them at -80°C for later use.

[0063] (2) Observation by electron microscope: after negative staining of the diseased juice, observe the morphology of virus particles by electron microscope.

[0064] (3) Purification of virus: Utilize the separation method of differential centrifugation and sucrose step density centrifugation to separate and purify the related virus of infected leaves.

[0065] (4) Identification of virus species: RT-PCR was used to detect and identify the virus species of Narcissus.

[0066] 2), the preparation of culture medium, including each component of basic culture medium and culture medium of each stage of tissue culture and the contained weight per liter are:

[0067] (1) Basic medium: select MS medium, sucrose or white sugar 20-40g / L, agar 7-9g / L, pH 5.6-5.8;

[0068] (2) D...

Embodiment 2

[0103] 1) Identification of the pathogenic species of Narcissus virus

[0104] (1) Take disease samples: take the disease samples of daffodil mosaic and leaf disease, and store them at -80°C for later use.

[0105] (2) Observation by electron microscope: after negative staining of the diseased juice, observe the morphology of virus particles by electron microscope.

[0106] (3) Purification of virus: Utilize the separation method of differential centrifugation and sucrose step density centrifugation to separate and purify the related virus of infected leaves.

[0107](4) Identification of virus species: RT-PCR was used to detect and identify the virus species of Narcissus.

[0108] 2), the preparation of culture medium, including each component of basic culture medium and culture medium of each stage of tissue culture and the contained weight per liter are:

[0109] (1) Basic medium: select MS medium, sucrose or white sugar 20-40g / L, agar 7-9g / L, pH 5.6-5.8;

[0110] (2) Da...

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PUM

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Abstract

The invention relates to a method for preserving tissue culture bulbs of narcissus l viruses, which belongs to the technical field of plant virus preservation and comprises following steps: identifying pathogeny species of the viruses, preparing culture medium, culturing virus-free tissue culture bulbs, testing virus ELISA, vaccinating virus by friction, and preserving virus bulbs. The invention has the advantages that the tissue culture bulbs of the narcissus l viruses are used to infect the purified viruses, and then the viruses of narcissus l tissue culture bulbs are preserved, which overcomes the defects in traditional preservation, can prevent viruses from confounding and losing, is easy to extract and prepare infectious titer, provides virus with high purity for preparing antiviral breeding and antiviral serum, lowers preserving cost, increases preserving effects, and is easy to carry and communicate.

Description

technical field [0001] The invention relates to the technical field of plant virus-free tissue culture and rapid propagation, in particular to a method for preserving tissue culture bulbs of narcissus virus. Background technique [0002] Narcisses (Narcissus) is a general term for new varieties of narcissus introduced into my country from Europe, and it is a perennial herb of the genus Narcissus in the family Amaryllidaceae. After the 1980s, large-scale introductions were carried out, mainly for spring flower exhibitions, and a small amount of potted plants were used as New Year's Eve flowers. Compared with Chinese narcissus, foreign narcissus has the characteristics of large flowers, bright colors, and pleasant fragrance; most varieties have trumpet-shaped corollas, and most of them are yellow, so they are also called "trumpet narcissus" or "yellow narcissus". Narcissus is an excellent garden ground cover flower, which can be used as potted ornamental and cut flowers. Daf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12Q1/70C12Q1/68
Inventor 陈剑平徐刚汪一婷吕永平牟豪杰郑红英林林程晔
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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