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Tobacco mosaic virus (TMV) resistant bacillus amyloliquefaciens

A technology of amylolytic spores and mosaic viruses, applied in the direction of bacteria, disinfectants, microbial-based methods, etc.

Inactive Publication Date: 2014-07-23
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no high-quality and high-resistance variety that can be widely promoted

Method used

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  • Tobacco mosaic virus (TMV) resistant bacillus amyloliquefaciens
  • Tobacco mosaic virus (TMV) resistant bacillus amyloliquefaciens
  • Tobacco mosaic virus (TMV) resistant bacillus amyloliquefaciens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Take 500g of tobacco root soil 5cm below the topsoil from the tobacco base of Hunan Agricultural University in Changsha City, Hunan Province, weigh 5g of the test soil and put it into a triangular flask filled with 45ml sterile water. After shaking for 1h, incubate in a constant temperature incubator at 28°C Let stand overnight. Remove the precipitate by suction filtration, which is 10 -1 bacterial suspension. Dilute the bacterial suspension by 10 with sterile water -2 、10 -3 、10 -4 、10 -5 、10 -6 After performing gradient dilution, spread them in NA medium respectively, and culture them upside down in a constant temperature incubator at 28°C for 1 day, observe the growth of the colonies, and pick different single colonies in time to streak and purify for 2-3 times to obtain the strain of the present invention, - Store at 80°C.

Embodiment 2

[0022] Example 2 Inhibition of bacterial strain Z5 of the present invention to tobacco common mosaic virus

[0023] Pick bacterial strain Z5 of the present invention and inoculate in 25ml NB substratum with 4% inoculum weight, 32 ℃, 180r / min cultivates 64h, gained fermented liquid removes thalline at 10500r / min centrifugation 15min, bacteria filter (0.45um) filters to obtain Sterile fermentation broth.

[0024] Take 0.1 g of TMV susceptible dry leaves and grind it into a homogenate, add PBS buffer solution at a ratio of 1:200 (W / V) to prepare a 20-fold virus inoculum. Select healthy tobacco leaves at the 4-6 leaf stage for friction inoculation, the left half leaf is inoculated with equal volume mixture of NB medium and virus as control group, and the right half leaf is inoculated with sterile fermentation liquid mixed with equal volume of virus liquid as treatment group (the mixing time is 10 minutes), spray the leaves with water 5 minutes after inoculation. Inoculate 3-5 le...

Embodiment 3

[0030] Example 3 Anti-TMV Activity Detection of Fermentation Supernatant Crude Protein of Bacterial Strain Z5 of the Present Invention

[0031] According to 100:56.1 (V / W), slowly add ammonium sulfate to the sterile fermentation broth in an ice-water bath, stir while adding, precipitate the protein in the fermentation broth, let stand at 4°C for 12h, 8000r / min, centrifuge at 4°C for 15min , PBS to dissolve the precipitate, then place it in a dialysis bag (molecular weight cut-off: 8000-14000), and dialyze to remove salt to obtain a crude protein solution (because it is the method of using conventional ammonium sulfate to precipitate protein, after adding ammonium sulfate, the substance is separated out in the fermentation broth , which means that the precipitated substance should be crude protein).

[0032] The dry leaves infected with TMV were treated with the above-mentioned crude protein liquid, sterile fermentation liquid and deproteinized liquid respectively, and the resu...

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Abstract

The invention provides a tobacco mosaic virus (TMV) resistant bacillus amyloliquefaciens strain Z5. The strain was collected in the China Center for Type Culture Collection (CCTCC) on March 24, 2014, with the collection number of CCTCC NO:M2014102. Fermentation liquor of the strain Z5 has inhibiting effects on the TMV. The in-vitro TMV inhibition ratio of the aseptic fermentation liquor is measured to be above 96% by utilizing a local lesion method. Coarse protein liquor prepared by precipitating the fermentation liquor has inhibiting effects on the TMV as well, has good antivirus effects and obvious control effects in the fields and can be used for effectively reducing the occurrence of TMV.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a Bacillus amyloliquefaciens resistant to tobacco common mosaic virus, in particular to a Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) Z5 strain, and at the same time relates to an antagonistic crude protein component obtained from the fermentation liquid of the strain Application in the control of tobacco common mosaic virus. Background technique [0002] Tobacco mosaic virus (TMV) is the most widely distributed and common disease in tobacco production so far. There are 16 kinds of tobacco virus diseases that have been found in my country, among which 3 kinds of viruses, TMV, CMV and PVY, are the main viruses that cause tobacco mosaic disease. They can parasitize and replicate in plant cells, and the materials, energy and places they need also come from the host. After tobacco is infected with the virus, chlorophyll is destroyed, photosynthesis is weakened, leaf growth is...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02A01N63/00A01P1/00C12R1/07
Inventor 唐前君周鹏刘双清肖启明李魏周志成李迅
Owner HUNAN AGRICULTURAL UNIV
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