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Method for removing strawberry light yellow edge virus by ultra low temperature technique

A light yellow edge virus and ultra-low temperature technology, applied in the field of bioengineering, can solve the problems of cumbersome operation and low detoxification rate, and achieve the effect of improving the detoxification rate and the detoxification efficiency.

Inactive Publication Date: 2009-04-29
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

Therefore, solving the problem of strawberry virus disease has become the focus of future research
The purpose of the present invention is to study the method of using ultra-low temperature to remove plant virus in view of the current situation that the current traditional detoxification method has a low detoxification rate in plant virus detoxification and the operation is too cumbersome.

Method used

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  • Method for removing strawberry light yellow edge virus by ultra low temperature technique
  • Method for removing strawberry light yellow edge virus by ultra low temperature technique
  • Method for removing strawberry light yellow edge virus by ultra low temperature technique

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Embodiment Construction

[0039] 1) Using the strawberry cultivar 'Mingbao' as the material, refer to the mild yellow edge disease symptoms of strawberries described by Wang Guoping (Chinese Fruit Tree Virus Pathogen Color Atlas [M]. Beijing: Golden Shield Publishing House. 2000), and look for leaf edge chlorosis in the field , dwarfed strawberry plants with yellowed and curled leaves. The taken plants are carried out total RNA extraction, reverse transcription is cDNA, according to the SMYEV coat protein sequence (accession number D12517) and ndh B gene sequence (accession number AJ316582), with reference to the research design primers such as Thompson (Thompson J R, Wetzel S, Klepks M.M, D, Schoen C.D, J, Jelkmann W. Multiplex RT-PCR detection of four aphid-borne strawberry viruses in Fragariaspp. in combination with a plant mRNA specific internal control [J]. Journal of Virological Methods, 2003, 111(2): 85-93.).

[0040] The primers for ndh B are:

[0041] P1: 5′-GGACTCCTGACGTATACGAAGGATC-3′ ...

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Abstract

The invention relates to 'a method for removing strawberry mild yellow edge virus (SMYEV) by the ultralow temperature technique', which belongs to the field of bioengineering. During the virus removal process, the concentration of pre-cultured sucrose is 0.5 mol per liter, and 3d is treated; the 3d is subjected to loading treatment for 60 minutes at a temperature of 25 DEG C; the 3d is subjected to vitrification treatment for 120 minutes at a temperature of 0 DEG C; and the 3d is subjected to liquid nitrogen treatment for 60 minutes and water bath treatment for 2 minutes at a temperature of 40 DEG C, wherein the survival rate of strawberry stem tips is 76 percent, and the detoxication rate of the strawberry mild yellow edge virus reaches 95 percent. The virus can be only removed by means of the liquid nitrogen treatment, and the detoxication rate of the vitrification treatment before the liquid nitrogen treatment is 0 percent. Compared with the prior detoxication method such as the stem tip culture detoxication method, the heat treatment detoxication method, and so on, the ultralow temperature detoxication method not only has quite high detoxication rate, but also is simple and feasible, convenient to operate, and does not require expensive instruments, so as to provide strong technical support for demonstration and popularization and industrialization of nontoxic strawberry seedlings.

Description

(1) Technical field [0001] The "method for removing strawberry mild yellow edge virus (SMYEV) by ultra-low temperature technology" of the present invention belongs to the field of bioengineering, and the method can effectively remove strawberry mild yellow edge virus. (2) Background technology [0002] 1 The principle of ultra-low temperature removal of virus [0003] The method of ultra-low temperature removal of plant virus is based on ultra-low temperature preservation combined with tissue culture and virus detection technology to achieve the purpose of detoxification. The combination of cryopreservation and shoot tip culture is an effective method for plant detoxification and preservation of shoot tips. The apical cells containing the virus are larger, the vacuoles are also larger, and the vacuoles contain more water, which are easily damaged and killed by ice crystals formed during ultra-low temperature storage, while the meristems with a faster proliferation rate cont...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68A01H4/00
Inventor 张计育渠慎春蔡斌华高志红乔玉山陶建敏徐长宝章镇
Owner NANJING AGRICULTURAL UNIVERSITY
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