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A Tobacco Common Mosaic Virus Biocontrol Endophyte Alcaligenes faecalis Strain

A technology of mosaic virus and Alcaligenes faecalis, which is applied in the direction of bacteria, biocides, microorganisms, etc., can solve the problems that there are few reports of inhibition and no reports of inhibition of tobacco endophytes.

Active Publication Date: 2018-08-31
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] Domestic reports on the biological control of tobacco common mosaic virus disease, the screening sources of bio-control bacteria are mostly from the rhizosphere soil, rhizosphere and other microbial flora, and the reports on the inhibitory effect of bio-control bacteria from inside plants on TMV Fewer, but no report on the inhibitory effect of tobacco endophytes on TMV

Method used

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  • A Tobacco Common Mosaic Virus Biocontrol Endophyte Alcaligenes faecalis Strain
  • A Tobacco Common Mosaic Virus Biocontrol Endophyte Alcaligenes faecalis Strain
  • A Tobacco Common Mosaic Virus Biocontrol Endophyte Alcaligenes faecalis Strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Screening of TMV antagonistic endophytic strain L1

[0045] Tobacco strains of TMV resistant varieties were taken from the experimental base of the Tobacco Research Institute of the Chinese Academy of Agricultural Sciences in Jimo City, Shandong Province, and 1.0 g pieces were cut from the tobacco leaves with a sterile knife, and the surfaces were disinfected with 70% ethanol and 1% sodium hypochlorite. The sterile water for the last rinse after the tissue surface disinfection was used as a blank control to test the surface disinfection effect. In the aseptic workbench, put the sterilized sample into a sterilized mortar and grind. Dilute the slurry with sterile water to 1×10 -4 , 1×10 -5 , 1×10 -6 , 1×10 -7 , 1×10 -8 concentration. Use a pipette to draw 100 μL of each dilution slurry and spread it on the enriched medium plate, place it in a 28°C constant temperature incubator for 3 days, select different single colonies and streak and purify 3 times again. The s...

Embodiment 2

[0046] Example 2 Identification of strain L1 of the present invention

[0047] 2.1 Physiological and biochemical identification of strain L1 of the present invention

[0048] The growth status of strain L1 after streaking on the LB medium plate figure 1 .

[0049] The colony is milky white and opaque, with neat edges, smooth surface and no water-soluble pigments.

[0050] The morphology of strain L1 under the transmission electron microscope figure 2 .

[0051] For the preparation of negative-stained specimens, a loop of Alcaligenes faecalis growing on the LB medium plate is used to prepare a bacterial suspension with sterile normal saline, and one drop is added to the copper mesh carrier membrane to be negatively stained with 1% phosphotungstic acid.

[0052] The negatively stained specimens were observed under an acceleration voltage of 80KV with H-600 transmission electron microscope. Critical point observation.

[0053] Strain L1 bacteria mostly exist as a single individual, the bac...

Embodiment 3

[0061] Example 3 Inhibition of the strain L1 of the present invention on tobacco common mosaic virus disease

[0062] 3-1 Preparation of fermentation broth: Pick a single colony that grows well on the LB medium plate and inoculate it into the LB liquid medium, culture it at 28° C., 180 rpm, for 24 hours for activation. Then, the activated bacterial liquid is inserted into the LB liquid medium with an inoculum of 2% for fermentation culture, and then the fermentation liquid can be obtained. The culture conditions are: temperature 28°C, pH 7.0, time 22h, rotating speed 180rpm.

[0063] 3-2 Detection method: Using the biological assay method of inoculating dead spot host Sansheng-NN tobacco to determine the inhibitory effect of the fermentation broth on tobacco common mosaic virus (TMV). Take the upper tender leaves of TMV tobacco that have fully diseased, grind them into a homogenate with a sterile mortar, and dilute with sterilized phosphate buffer (PH7.0) at a ratio of 1:80. Sele...

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Abstract

The invention discloses an endophyte strain Alcaligenes faecalis used for biocontrol of the tobacco mosaic virus. The strain has a code of L1 and is preserved in China General Microbiological Culture Collection Center on October 13, 2014, with an accession number of CGMCC No. 9758. The strain can inhibit invasion of the tobacco mosaic virus; biocontrol experiments show that the strain can prevent and treat the tobacco mosaic virus; and intracellular crude protein extract capable of preventing and treating the tobacco mosaic virus is extracted from the strain, and the crude protein extract has a high application value in prevention and treatment of the tobacco mosaic virus.

Description

Technical field [0001] The present invention belongs to the field of biological technology, and relates to a strain of Alcaligenes faecalis, an endophytic endophyte of tobacco common mosaic virus, in particular to a strain of Alcaligenes faecalis L1, and also to a cell obtained from the strain. The method of internal crude protein composition and its application in tobacco common mosaic virus control. Background technique [0002] China is a large tobacco planting country, and its tobacco planting area and output rank first in the world. At the same time, tobacco is also one of the main economic crops in my country. It is grown in more than 20 provinces and regions across the country. Yunnan, Guizhou and Sichuan are the major tobacco growing provinces in my country. [0003] However, the occurrence and prevalence of various tobacco pests and diseases have caused huge economic losses while reducing the yield and quality of tobacco. Among them, tobacco virus causes serious damage a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/02A01N63/00A01P1/00C12R1/05
Inventor 战徊旭王凤龙申莉莉杨金广钱玉梅陈德鑫刘伟刘旭
Owner TOBACCO RES INST CHIN AGRI SCI ACAD
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