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A strain of Bacillus subtilis and its application in the control of eggplant brown spot disease

A technology for bacillus subtilis and eggplant brown spot disease, applied in the fields of application, bacteria, and fungicides, can solve the problems of the five major circles of the earth's surface system and the increase in the degree of drug resistance of bacteria

Active Publication Date: 2021-12-21
WEIFANG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, when the disease occurs, chemical pesticides are still used to kill the bacteria, but the long-term use of chemicals not only increases the drug resistance of the bacteria, but also causes non-negligible harm to the five major circles of the earth's surface system

Method used

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  • A strain of Bacillus subtilis and its application in the control of eggplant brown spot disease
  • A strain of Bacillus subtilis and its application in the control of eggplant brown spot disease
  • A strain of Bacillus subtilis and its application in the control of eggplant brown spot disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Purification of Bacillus, preliminary identification

[0033] 1, the method: Soil samples were collected from different parts of the starch-containing medium, 37 [deg.] C culture sample coated 1-3d, characterized in colony morphology was observed using a soil dilution method. Picked off-white corrugated edge suspected colonies were inoculated onto agar plates BPY, 37 ℃ pure culture 48h. Using Gram stain, catalase reaction, V-P reaction, the reaction amylase preliminary identification of bacteria classification.

[0034] Specifically, 10% hydrogen peroxide is injected directly into the inclined surface, whether or not air bubbles observed. Bubbles were positive, negative reaction inaction.

[0035] In particular, isolates taken broth and mixed equal amounts of 40% NaOH, add a little creatine (about 0.5-1.0 mg), shaken vigorously, after 2-10min Observe for red, a red if the VP was positive .

[0036] In particular, isolates of the culture was taken starch biochemica...

Embodiment 2

[0040] Bacillus 16S-rDNA sequence of Example 2 Amplification and Analysis embodiment

[0041] 1. Method:

[0042] Subtilis strain using the obtained initial identification, inoculated into LB liquid medium at 37 ℃ constant temperature incubator, 200rpm / min shaker 24h. Then suction was centrifuged broth is repeated until a certain amount of deposit, using bacterial DNA extraction kit (OMEGA) Extraction of Bacillus total DNA.

[0043] Using bacteria 16SRNA amplification primers on DNA extracted from total Bacillus PCR amplification, the product obtained was sequenced. Sequencing results homology alignment by Blast.

[0044] Bacterial 16SRNA amplification primers comprises Eubac27F and Eubac1492R;

[0045] Wherein the nucleotide sequence of primer Eubac27F as shown in SEQ ID NO 1, in particular:

[0046] 5'-AGAGTTTGATCCTGCCTCAG- 3 ';

[0047] Wherein the nucleotide sequences of primers Eubac1492R as shown in SEQ ID NO 2, in particular:

[0048] 5'-GGATACCTTGTTACGACTT-3 '.

[0049] ...

Embodiment 3

[0054] Example 3 Isolation, activation and morphological identification of eggplant brown pathogenesis

[0055] 1, method: from the field, a disease with a typical eggplant browning disease is collected, and the strain is separated from the tissue separation method (reference: Li Yanqing, etc., Shandong Shouguang Eggplant Brown Pathogenic Identification and Compaction [J]. Northern Gardening, 2017 (9): 106-110). In the sterile operating table, in the PDA plate inoculation, pay attention to after inoculating, it is necessary to sterilize the pick-up needle, and then placed in a 25 ° C electrothermal incubator, 7-14 days, and the colony is long.

[0056] Picking the spore is placed on the slide and is released by extrusion, and the spore is released and observed under an optical microscope. According to Yongli, etc. in 1998, "Percutaneous Physicia in Biocide, I: Class I: Shell Division, etc. PHomopsis Description, referred to the fungal identification manual (Wei Jingchao, 1979) and...

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Abstract

The invention discloses a strain of Bacillus subtilis, the preservation number of which is CCTCC NO: M 2021862, which is preserved in the China Center for Type Culture Collection. The invention also discloses the application of the above bacillus subtilis in the prevention and treatment of eggplant brown spot disease. Specifically, it is applied to inhibit the growth of Phytophthora solani. It is verified by confrontation culture experiments that the Bacillus subtilis of the present invention has a high-efficiency and broad-spectrum inhibitory effect on the brown spot disease of eggplant, and the inhibition rate of different strains of the brown spot disease of eggplant is more than 50%, which can be applied to the prevention and treatment of brown spot disease of eggplant .

Description

Technical field [0001] The present invention belongs to the field of biotechnology, particularly relates to a Bacillus subtilis and Its Application in eggplant Phomopsis rot Control. Background technique [0002] Eggplant Phomopsis rot disease is one of the three growth process eggplant, tomato Phomopsis by Phomopsis ( PHomopsis vexans Harter ) A soil-borne diseases caused by bacteria, is a worldwide disease. China discovered eggplant brown grain disease as early as 1932 (Liu Xuemin et al, 1998), was first reported in Gujarat in India, currently the disease and its related hazards Solanaceae plants across the country. Currently, when the disease occurs is still using chemical pesticides to kill germs, but long-term use of chemicals not only increase the level of resistance of bacteria on the earth surface system of five rings also produces hazards can not be ignored. Now with biocontrol agents to control diseases gradually become the focus of attention of domestic and foreign res...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P3/00C12R1/125
CPCA01N63/22
Inventor 李艳青朱秀高潘好芹李春雷袁旭超王宗伟
Owner WEIFANG UNIV OF SCI & TECH
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