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Biological control pseudomonas monteilii strain against tobacco mosaic virus (TMV)

A technology of Pseudomonas monsonii and mosaic virus, which is applied in the direction of bacteria, microorganisms, and methods based on microorganisms, can solve the problems that the mechanism of physiological and biochemical characteristics of disease prevention has not been studied in depth, so as to prevent virus transmission, promote growth, Inhibition of infection and proliferation

Active Publication Date: 2013-09-04
TOBACCO RES INST CHIN AGRI SCI ACAD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] There have been reports on the biological control of common tobacco mosaic virus disease in China, but most of them are limited to the screening of bio-control bacteria and pot experiments, and the extraction of antibacterial substances, physiological and biochemical characteristics and disease prevention mechanism of bio-control bacteria, etc. Many aspects have not been studied in depth in China

Method used

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  • Biological control pseudomonas monteilii strain against tobacco mosaic virus (TMV)
  • Biological control pseudomonas monteilii strain against tobacco mosaic virus (TMV)
  • Biological control pseudomonas monteilii strain against tobacco mosaic virus (TMV)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1.1 Screening of TMV antagonistic bacteria 4A1 strain

[0029] Pick the purified strain and inoculate it in LB liquid medium, keep the constant temperature at 28°C, and shake at 140rpm for 48 hours, take 20ml and mix it with an equal volume of 40 times TMV juice for 15 minutes, rub the TMV-inoculated rotten spot host Sansheng-NN tobacco, and inoculate the upper part of each strain For 2 leaves, use the mixture of sterile water and TMV as a control, and investigate the dead spot inhibition rate for 3 to 5 days. The strains with antagonistic activity in the preliminary test were tested repeatedly in the same way, with 3 strains for each treatment, and three repetitions.

[0030] Calculation formula of bruise inhibition rate:

[0031]

[0032] The investigation of rot inhibition rate showed that strain 4A1 had the best inhibitory effect on TMV, and the rot inhibition rate of TMV reached 98.0%. Biocontrol bacteria produced antibiotic substances with anti-virus activity ...

Embodiment 2

[0045] Observation of Effect of Active Substances of 4A1 Strain on TMV Virus Particles under the Perspective Electron Microscope

[0046] (1) Rough purification of TMV granules

[0047] Purify TMV mitochondria with reference to the PEG method (Hu Weizhen et al., 1989), and the specific operation steps are as follows:

[0048] Add 100g of live TMV poison source leaves, add the same amount of sterile water, fully grind until homogenized, filter with nylon gauze, centrifuge at 1000rpm for 10min, and discard the precipitate;

[0049] The supernatant was heated at 60°C for 10 minutes in a water bath, centrifuged at 4000 rpm for 20 minutes, and the precipitate was discarded;

[0050] Add 4% PEG to the supernatant 6000 (0.1M NaCl) or 2% PEG 6000 (0.3M NaCl), discard the supernatant;

[0051] The resulting precipitate is TMV purified virus.

[0052] (2) Preparation and observation of negative stain electron microscope samples

[0053] The 4A1 fermentation supernatant was mixed w...

Embodiment 3

[0059] 3.1 Comparison of the effect of strain 4A1 with other antiviral agents

[0060] Choose the virus inhibitors that are often used in production: Ningnanmycin (200 times), morpholinoguanidine (400 times), soap water (20%) and the comparison of the antiviral effects of 4A1. Each virus inhibitor was evenly mixed with 40 times the virus juice for 15 minutes, and the mixture of sterile water and TMV was used as a control, and the half-leaf method was inoculated on Sansheng-NN tobacco, and the upper 2 leaves of each plant were inoculated, 3 plants per treatment, three times Repeat, 3 ~ 5d to investigate the suppression of dead spots. See Table 3 for details.

[0061] The anti-disease effect of table 3 multiple virus inhibitors

[0062]

[0063] As can be seen from Table 3, the commonly used antiviral agents morpholinoguanidine and Ningnanmycin have a control effect of less than 70%, the inhibition rate of soapy water is only 11.6%, and the virus inhibition rate of 4A1 has ...

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Abstract

The invention discloses a biological control pseudomonas monteilii strain against tobacco mosaic virus (TMV). The code name of the strain is 4A1, the strain is collected in China General Microbiological Culture Collection Center (CGMCC) on September 7, 2011, and the collection number of the strain is CGMCC No. 5229. The strain can inhibit the infection of the TMV, and a biological control experiment shows that the strain can control the TMV. Simultaneously, the invention further provides a method for separating active substances from fermentation supernatant fluid obtained from the strain, and the active substances have a very high application value in the aspect of controlling the TMV.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a strain of Pseudomonas monsonii for the biocontrol of tobacco common mosaic virus, the bacterial strain and the use of isolated active substances in the fermentation supernatant obtained from the strain in the prevention and treatment of tobacco common mosaic virus application. Background technique [0002] Tobacco mosaic virus disease is caused by tobacco mosaic virus (TMV). The most notable symptom is the formation of yellow and green mottled mosaic leaves, and the edges of diseased leaves sometimes curl to the back. The disease can occur continuously throughout the growth period from the seedbed to the field, and the color of the diseased leaves is uneven after modulation, the taste is poor, and the quality declines. Tobacco common mosaic virus is widely distributed in various smoking areas in my country, and Heilongjiang, Jilin, Liaoning, Shandong, Henan, Anhui, Guangdong and oth...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/38
Inventor 王凤龙钱玉梅杨金广申莉莉翟熙伦陈德鑫孙惠青王新伟战徊旭
Owner TOBACCO RES INST CHIN AGRI SCI ACAD
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