Endophyte strain Alcaligenes faecalis used for biocontrol of tobacco mosaic virus

A technology of mosaic virus and Alcaligenes faecalis, applied in directions such as bacteria, biocides, microorganisms, etc., can solve the problems such as no reports on the inhibitory effect of tobacco endophytes, few reports on the inhibitory effect, etc.

Active Publication Date: 2015-12-02
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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  • Endophyte strain Alcaligenes faecalis used for biocontrol of tobacco mosaic virus
  • Endophyte strain Alcaligenes faecalis used for biocontrol of tobacco mosaic virus
  • Endophyte strain Alcaligenes faecalis used for biocontrol of tobacco mosaic virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Prevention and treatment of TMV against the screening of endophyte strain L1

[0045] Tobacco plants of TMV-resistant varieties were taken from the experimental base of Tobacco Research Institute of Chinese Academy of Agricultural Sciences in Jimo City, Shandong Province, and 1.0 g small pieces were cut from the leaves of the tobacco plants with a sterile knife, and the surface was sterilized with 70% ethanol and 1% sodium hypochlorite. The sterile water after the last flushing of the tissue surface after disinfection was used as a blank control to detect the surface disinfection effect. In an aseptic bench, grind the sterilized sample in a sterile mortar. Dilute the grinding liquid with sterile water to 1 × 10 -4 , 1×10 -5 , 1×10 -6 , 1×10 -7 , 1×10 -8 concentration. Use a pipette gun to draw 100 μL of each dilution grinding solution and spread it on the enrichment medium plate, place it in a constant temperature incubator at 28°C and cultivate it for 3...

Embodiment 2

[0046] The identification of embodiment 2 bacterial strain L1 of the present invention

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

[0048] The growth state of strain L1 after streaking on LB medium plate is shown in figure 1 .

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

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

[0051] Negative staining specimen preparation, take a ring of Alcaligenes faecalis grown on the LB medium plate, use sterile saline to make a bacterial suspension, add one drop to the copper mesh support membrane, and negatively stain with 1% phosphotungstic acid.

[0052] Negatively stained specimens were observed with an H-600 transmission electron microscope at an accelerating voltage of 80KV. Critical point observation.

[0053] Strain L1 cells mostly exist as a sin...

Embodiment 3

[0061] Embodiment 3 Inhibitory effect of bacterial 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 at 28°C, 180rpm, for 24h for activation. Then the activated bacterial liquid is inserted into LB liquid culture medium with an inoculum amount of 2% to carry out fermentation culture to obtain a fermented liquid. The culture conditions are as follows: temperature 28° C., pH 7.0, time 22 hours, rotation speed 180 rpm.

[0063] 3-2 Detection method: The inhibitory effect of the fermentation broth on tobacco mosaic virus (TMV) was measured by using the biological assay method of inoculating Sansheng-NN tobacco, the host of rotten spots. The upper tender leaves of TMV tobacco that are fully infected are taken, ground into a homogenate with a sterile mortar, and diluted with sterilized phosphate buffe...

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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 invention belongs to the field of biological technology, and relates to a tobacco common mosaic virus biocontrol endophyte Alcaligenes faecalis strain, in particular to a Alcaligenes faecalis L1 strain, and at the same time relates to the intracellular The method of crude protein composition and its application in the control of tobacco common mosaic virus. Background technique [0002] China is a big tobacco planting country, with the tobacco planting area and output ranking first in the world. At the same time, tobacco is also one of the main economic crops in my country, and it is planted in more than 20 provinces and regions across the country. Yunnan, Guizhou and Sichuan are the major provinces of tobacco planting in my country. [0003] However, the occurrence and prevalence of various tobacco diseases and insect pests have caused huge economic losses while reducing the yield and quality of tobacco. Among them, the cause of tobacco virus is...

Claims

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Application Information

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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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