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Bacillus subtilis for preventing and treating phytophthora blight of pepper

A technology of Bacillus subtilis and Pepper Phytophthora, applied in the direction of chemicals for biological control, methods based on microorganisms, bacteria, etc., can solve the problems of difficult to effectively control Pepper Phytophthora, adverse effects on health, and destruction of agricultural ecological balance, etc. problem, to achieve good suppression effect, short cycle and high safety effect

Inactive Publication Date: 2014-12-24
YANGTZE UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, chemical control is mostly used for the prevention and treatment of pepper blight, and the extensive use of chemical agents has caused serious environmental pollution, destroyed the ecological balance of agriculture, and adversely affected human health.
At the same time, because there are few pepper varieties resistant to blight, and most of them are medium-resistant or disease-resistant varieties, it is difficult to effectively control pepper blight.

Method used

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  • Bacillus subtilis for preventing and treating phytophthora blight of pepper
  • Bacillus subtilis for preventing and treating phytophthora blight of pepper
  • Bacillus subtilis for preventing and treating phytophthora blight of pepper

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Experimental program
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Effect test

Embodiment 1

[0031] Embodiment 1: Isolation, screening and identification of Bacillus subtilis BS04

[0032] (1) Separation and screening: Soil samples were collected from the rhizosphere of healthy peppers using the five-point sampling method from fields with severe epidemics in pepper planting areas such as Wuhan, Jingzhou, Yichang, and Jingmen, Hubei. When sampling, shovel off the top 5cm soil layer, take 500g of fresh soil samples, and bring them back with fresh-keeping bags.

[0033] Weigh 10 g of the soil sample and add it to a conical flask filled with 90 mL of sterile water. The conical flask is filled with a number of glass beads, vibrate at 28 °C and 120 r / min for 30 min, and obtain a soil suspension after standing still.

[0034] Dilute the soil suspension to 10 -5 、10 -6 、10 -7 100 μL of soil suspension with different dilutions was drawn and spread on NA medium, cultured at 28°C for 48 hours, and single colonies of bacteria with different colors and shapes were picked and pu...

Embodiment 2

[0047] Sequence comparison of the sequencing results on NCBI Genebank (comparison number: lcl|28017), found that its similarity with the 16S rDNA sequence of Bacillus subtilis was as high as 99%, and the antagonistic strain BS04 was preliminarily determined to be Bacillus subtilis (Bacillus subtilis) . Cluster analysis was performed using MEGA4.1 software, and a phylogenetic tree was constructed. The results showed that B. subtilis (KF725636) was in a small branch with B. subtilis (KC506778.1), and was in a large branch with the other two Bacillus subtilis. Thus, combined with the morphological and physiological and biochemical characteristics, it was further demonstrated that the BS04 strain was Bacillus subtilis. Embodiment 2: the antibacterial spectrum determination of bacillus subtilis BS04 bacterial strain

[0048] Bacillus subtilis BS04 strain against pepper blight (Phytophthora capsici), tobacco black shank (P.parasitica var.nicotiana), cotton Verticillium dahliae (Vert...

Embodiment 3

[0054] Embodiment 3: the disease prevention mechanism of bacillus subtilis BS04 bacterial strain to capsicum blight

[0055] (1) Effect of BS04 strain on the mycelial growth of Phytophthora capsici

[0056] After inoculating and culturing Phytophthora capsici on PDA medium for 3 days, a 5 mm diameter bacterial block was punched out and placed in a sterile petri dish filled with 20 mL PD medium, and cultured at 28°C for 2 days. Treat the cultured Phytophthora capsici bacterial block with the sterile culture filtrate and bacterial suspension of the BS04 strain respectively for 24 hours, and use sterile water treatment as the control, pick the hyphae and observe its morphology under an inverted microscope, and repeat each treatment. 3 times.

[0057] The test results showed that after the Phytophthora capsici was treated with the bacterial suspension of strain BS04 and the sterile filtrate, the mycelium branches increased, the top appeared deformed, the growth was slow, and the ...

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Abstract

The invention relates to bacillus subtilis BS04 for preventing and treating phytophthora blight of pepper, belonging to the technical field of biological prevention and treatment on agricultural diseases. The bacillus subtilis BS04 is characterized by being separated from rhizosphere soil of pepper, and being preserved in the China Center for Type Culture Collection (CCTCC) with the preservation number of CCTCC NO: M2014294. The bacillus subtilis BS04 has a bacteriostasis for pathogenic fungi such as pepper phytophthora blight, phytophthora nicotiana, cotton verticillium wilt virus, fusarium oxysporum vasinfectum, sclerotinia sclerotiorum and lettuce botrytis cirerea, the antibacterial rate for original bacteria is within 40-80%, and the antibacterial rate for pepper phytophthora blight is greater than 80%. The bacillus subtilis BS04 provided by the invention is obtained by being separated from soil of healthy plant roots, is free of ecotoxicity, high in security, wide in antimicrobial spectrum, simple to culture and short in period, and has very wide application prospect in biological prevention and control of plant diseases.

Description

Technical field: [0001] The invention relates to a bacillus subtilis used for preventing and treating pepper blight, and belongs to the technical field of biological control of agricultural diseases. Background technique: [0002] Pepper blight, commonly known as "dead seedling disease", is caused by Phytophthora capsici and is one of the main pepper root diseases. It occurs seriously in pepper production areas in my country and other countries in the world, causing huge economic losses to pepper production. . [0003] Phytophthora capsici is usually distributed in the soil, with strong pathogenic variation and easy to develop drug resistance. At present, chemical control is mostly used in the prevention and control of pepper blight, and the extensive use of chemical agents has caused serious environmental pollution, destroyed the ecological balance of agriculture, and adversely affected human health. At the same time, it is difficult to effectively control pepper blight be...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01P3/00C12R1/125
Inventor 孙文秀李伟
Owner YANGTZE UNIVERSITY
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