Salt-tolerance bacillus E40207a2 and application thereof

A technology of e40207a2 and halotolerant bacillus, applied in the field of microorganisms, to achieve effective antagonism and control, environmental safety, and good control effects

Active Publication Date: 2019-05-31
QINGHAI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on the use of halophilic bacteria strains to carry out biological control of potato virus diseases, especially the research and development of potato virus Y antiviral agents and the mechanism of action using such microbial resources is still a blank field

Method used

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  • Salt-tolerance bacillus E40207a2 and application thereof
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  • Salt-tolerance bacillus E40207a2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Isolation and Identification of Bacterial Strain E40207a2

[0033] 1) Isolation of strain E40207a2

[0034] Take an appropriate amount of salt lake mud samples, air-dry (about 7d), and treat at 120°C for 1h. Weigh 10g of the processed lake mud sample, add 90m L of sterile seawater, put it into a sterilized Erlenmeyer flask equipped with glass balls, shake it fully for 30min, and draw the supernatant after standing still, the supernatant is 10 -1 Soil sample. the above 10 -1 Samples were serially diluted to 10 -2 and 10 -3 times, pipette 150 μL of each gradient sample, spread it on each medium plate, and repeat 3 times for each sample. The coated plate was placed upside down in an incubator, and cultured upside down at 28°C and 37°C, respectively, and colonies were observed, and a single colony was picked and purified. The purified strains were stored on the slant of the test tube and placed in a refrigerator at 4°C. The medium is ATCC213 modified mediu...

Embodiment 2

[0041] Example 2 Strain Activation, Preparation of Fermentation Broth and Inhibition of PVY Resistance Evaluation

[0042] 1.1 Test medium, strain activation and fermentation broth preparation

[0043] 1.1.1 The halophilic bacteria activation fermentation medium is as follows:

[0044] ATCC213 modified medium: MgSO 4 ·7H 2 O 10g, CaCl 2 2H 2 O 0.2g, KCl 5g, peptone 2.5g, yeast extract 10g, NaCl 30g, agar powder 12g, distilled water to 1000mL, pH 7.2-7.4.

[0045] 1.1.2 The halophilic bacteria were isolated, purified and preserved from the Qinghai Chaerhan Salt Lake by the Key Laboratory of Qinghai-Tibet Plateau Biotechnology Ministry of Education. The preserved strains were streaked on the solid medium of ATCC213 modified medium, and cultured in a constant temperature incubator at 37°C for 24 hours for later use.

[0046] 1.1.3 Fermentation broth preparation: Inoculate the activated halophilic bacteria strain with 20% inoculum volume into a 1000mL Erlenmeyer flask with 8...

Embodiment 3 3

[0064] Example 3 Response expression of related stress-resistant genes in Sansheng Tobacco

[0065] 1) Template preparation

[0066] The total RNA extracted in Example 2 and the cDNA obtained by inversion were directly used as templates.

[0067] 2) Design of primers related to stress resistance genes in Sansheng Tobacco

[0068] Referring to the primer design method in Example 2, design the fluorescent primers of the relevant resistance genes in the host tobacco Sansheng tobacco (Table 3).

[0069] Table 3 Anti-stress gene sequence

[0070]

[0071] 3) Responsive expression of related stress-resistant genes in Sansheng Tobacco

[0072] One week after PVY infects Sansheng tobacco, the expression levels of 9 stress-resistant genes of the host Sansheng tobacco were quantitatively detected by Real-time PCR. Among them, glutathione-S-transferase (GST), PVY capsid protein interacting protein-3 (CpIp), Cullin protein-1 (Cullin), box family protein (F-Box) and eukaryotic trans...

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Abstract

The invention discloses salt-tolerance bacillus E40207a2 and application thereof. The salt-tolerance bacillus E40207a2 is preserved in Guangdong Microbiological Culture Collection Center on January third, 2019, and the preservation number is GDMCC NO:60498. A fermentation solution of a secondary metabolite product of a strain has an obvious antagonistic function on viruses and diseases caused by Potatovirus Y (PVY), and the material is wide in source, low in cost, and safe to the environment.

Description

technical field [0001] The invention belongs to the technical field of microbes, and relates to a halophilic bacterium derived from Chaerhan Salt Lake and its biocontrol application, in particular to a halotolerant bacillus E40207a2 and its application. Background technique [0002] Potato virus Y (PVY) is a typical member of the genus Potyvirus in the family Potyviridae. The virion is slightly curved and linear, with a length of 680-900nm and a width of 11-12nm. The genome of PVY is a single RNA positive strand with a relative molecular mass of 3.1×10 6 -3.5×10 6 , about 10,000 amino acids in length. At the 5' end of the genome is a covalently bonded genome-binding protein (VPg) and a non-coding region, about 180 bases; at the 3' end of the genome is a poly(A) tail, and the entire genome has only one reading frame. After translation, a large polyprotein is produced, which is processed by a self-encoded protease to form a mature coat protein and at least seven nonstructur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/02A01P1/00C12R1/07
Inventor 沈硕李玮宋宝安郭青云王舰
Owner QINGHAI UNIVERSITY
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