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Bacillus velezensis 05-1205 as well as acquisition method and application thereof

A technology of Bacillus Velez and Bacillus, applied in the field of microbial application, to achieve the effect of convenient application, low cost and good colonization effect

Active Publication Date: 2021-02-19
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there are few microbial agents used to prevent and treat mildew in the baking period, only Bacillus mucilaginosus (CN110384247A) and combined microbial agents (CN110250209A) are used as biocontrol products to prevent tobacco mildew in the curing period report

Method used

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  • Bacillus velezensis 05-1205 as well as acquisition method and application thereof
  • Bacillus velezensis 05-1205 as well as acquisition method and application thereof
  • Bacillus velezensis 05-1205 as well as acquisition method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] - Acquisition of Bacillus velezensis 05-1205

[0094] A. Separation: Take 0.4g of the stem of the tobacco plant that has been thoroughly sterilized on the surface after inspection, put it into a 2 mL sterilized centrifuge tube together with sterilized steel balls, add 600 μL of sterile water, and grind it with a tissue grinder for 3 minutes at a frequency of 30r / s. Gradually dilute the ground tissue fluid to 10 -4 , take 200 μL homogenate for each gradient and apply separation medium (peptone 5.0g / L, beef extract 3.0g / L, sodium chloride 8.0g / L, agar 15.0g / L, pH 7.05), each treatment is repeated 4 times . After dark culture at 28°C for 48 hours, pick a single colony according to the colony shape, color, etc., re-stretch and purify on the separation medium, and pick the single colony cultured by the streak and transfer it to the slant of the separation medium test tube for storage;

[0095] B. Screening: Using the flat plate confrontation method, stick the mycelia blo...

Embodiment 2

[0101] ——Identification and preservation of Bacillus velezensis 05-1205

[0102] (1) Identification of Bacillus velezensis 05-1205

[0103] For the above-mentioned strain 05-1205 selected and bred, biological and physiological and biochemical characteristics were detected and identified by molecular biological methods by conventional methods. The molecular identification method is as follows: Bacteria Genomic DNA Extraction Kit, for the method, refer to the kit instruction manual. Primer F27 / R1492 was selected for PCR amplification, amplified under conventional conditions, the amplified product was recovered by gel, connected to the vector pEAZY-T5 Zero vector, heat-shock transformed Escherichia coli competent cell DH5α, and colonies were picked with M13F / M13R as primers Perform colony PCR identification. Positive clones were sent to Shanghai Yingjun Biotechnology Co., Ltd. for sequence determination.

[0104] The test results are recorded as follows:

[0105] 1. Culture c...

Embodiment 3

[0114] ——The control test of Bacillus velezensis 05-1205 on tobacco mildew during curing period

[0115] The test was carried out in Fengyi Town, Xiaguan City, Dali Prefecture, Yunnan Province, and the test tobacco leaves were the middle tobacco leaves of the flue-cured tobacco variety "Honghua Dajinyuan".

[0116] experiment method:

[0117] a. Fermentation: activate the strain 05-1205 in a slant medium (peptone 10.0 g / L, beef extract 3.0 g / L, sodium chloride 5.0 g / L, agar 15.0 g / L, pH 7.0) from a -80°C ultra-low temperature refrigerator ), placed in a constant temperature incubator at 28°C for 48 hours; pick a ring of bacteria and inoculate it in 10mL seed culture solution (peptone 10.0g / L, beef extract 3.0g / L, sodium chloride 5.0g / L, agar 15.0g / L, pH 7.0) in a 50mL centrifuge tube, placed in a constant temperature incubator at 28°C for 24h; then the seed solution was transferred to 1000mL fermentation medium (peptone 10.0g / L, beef extract 3.0g / L, chlorine NaCl 5.0g / L, ag...

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Abstract

The invention discloses Bacillus velezensis 05-1205 as well as an acquisition method and application thereof, and belongs to the technical field of microbial application. The Bacillus velezensis 05-1205 is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number of the Bacillus velezensis 05-1205 is CCTCC No: M2020568. The acquisition method comprises processes of separating, screening and purifying, firstly, endophytic bacteria are seperated from healthy tobacco plants, endophytic bacteria antagonizing rhizopus oryzae are screened by taking the tobacco mildew pathogenic bacteria rhizopus oryzae in a baking period as an indicator, and finally, purifying is conducted on a culture medium plate to obtain the Bacillus velezensis 05-1205. The application ofthe bacterial strain comprises application in tobacco mildew control, the bacterial strain Bacillus velezensis 05-1205 can achieve effective control over tobacco mildew in the baking period, and therelative control effect is 70% or above. Secondly, the Bacillus velezensis 05-1205 is wide in antibacterial spectrum, and has a very good antibacterial effect on tobacco bacterial wilt bacteria, botrytis cinerea, sclerotinia sclerotiorum, target leaf spot bacteria, root rot bacteria and alternaria alternata besides pathogenic bacteria of tobacco mildew in the baking period.

Description

technical field [0001] The invention belongs to the technical field of microbial application, and in particular relates to a strain of Bacillus veleisi 05-1205, an acquisition method and its application. Background technique [0002] Tobacco curing loss is the loss caused by abnormal curing of tobacco leaves in the barn, which causes heavy losses to tobacco leaf production and tobacco farmers. In recent years, tobacco mildew during the curing period has become one of the important economic losses during the curing process in some southern tobacco-growing areas such as Fujian, Yunnan, and Guangxi. Tobacco mildew during the curing period is mainly caused by the infection of the tobacco leaves by Rhizopus oryzae due to the low temperature and high humidity during the yellowing period of the tobacco leaves. [0003] The prevention and control methods of mildew during the baking period mainly include the improvement of baking equipment, chemical control and biological control. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02A01N63/22A01P3/00C12R1/07
CPCC12N1/20C12N1/02A01N63/22C12R2001/07C12N1/205
Inventor 卢灿华夏振远晋艳姜宁盖晓彤马俊红雷丽萍苏家恩鲁子贤高朝阳蔡永占
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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