Tissue culture bulb preserving method for thunberg fritillary virus
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A kind of preservation method, technology of fritillary
Inactive Publication Date: 2008-07-09
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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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
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Embodiment 1
[0063] 1), the identification of the pathogenic species of Fritillaria virus
[0064] (1) Take disease samples: Take the disease samples of Fritillaria mosaic disease and store them at -80°C for later use.
[0065] (2) Observation by electron microscope: after negative staining of the diseased juice, observe the morphology of virus particles by electron microscope.
[0066] (3) Identification of virus species: RT-PCR was used to detect and identify the virus species of Fritillaria fritillata.
[0067] 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:
[0068] (1) Basic medium: select MS medium, sucrose or white sugar 20-40g / L, agar 7-9g / L, pH 5.6-5.8;
[0069] (2) Daughter bulb induction medium: MS+2, 4-D 0.5~1.5mg / L and ZT 1~3mg / L+ sucrose or white sugar 30g / L;
[0070] (3) Daughter bulb proliferation medium: MS+BA 1~3mg / L and NAA 1~3mg / L+sucrose...
Embodiment 2
[0106] 1), the identification of the pathogenic species of Fritillaria virus
[0107] (1) Take disease samples: Take the disease samples of Fritillaria mosaic disease and store them at -80°C for later use.
[0108] (2) Observation by electron microscope: after negative staining of the diseased juice, observe the morphology of virus particles by electron microscope.
[0109] (3) Identification of virus species: RT-PCR was used to detect and identify the virus species of Fritillaria fritillata.
[0110] 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:
[0111] (1) Basic medium: choose MS basic medium, 20-40g / L sucrose or white sugar, 7g / L agar, pH5.8;
[0112] (2) Daughter bulb induction medium: MS+2, 4-D 1mg / L and ZT 2mg / L+ sucrose 30g / L;
[0113] (3) Daughter bulb proliferation medium: MS+BA 2mg / L and NAA 2mg / L+white sugar 40g / L+thiamine hydrochl...
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Abstract
The invention relates to a method for preserving tissue culture bulbs of fritillary 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 fritillary viruses are used to infect the purified viruses, and then the viruses of fritillary 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 detoxification tissue culture rapid propagation, in particular to a method for cultivating the detoxification tissue culture bulbs of Fritillaria fritillaria. Background technique [0002] Fritillaria thunbergii Miq. (Fritillaria thunbergii Miq.) is a plant of the genus Fritillaria in the family Liliaceae. Its dried bulbs are one of the famous Chinese medicinal materials "Zhe Bawei". Zhejiang Fritillaria is native to Xiangshan County, so it is also called Elephant Fritillaria. It is said that during the Kangxi period of the Qing Dynasty, farmers in Xiangshan began to transfer Fritillaria from wild to home. For a long time, Pan'an and Yinzhou are the main producing areas of Fritillaria fritillaria, accounting for about 70% of the total output of Fritillaria fritillata in China. It is also cultivated in Jiangsu, Jiangxi, Shanghai, Hubei and Hunan provinces. With the in-depth adjustment of planting i...
Claims
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