CN110117552B

CN110117552B - Bacillus belgii and application thereof

Info

Publication number: CN110117552B
Application number: CN201810110929.8A
Authority: CN
Inventors: 仲乃琴; 宋双伟; 赵盼; 钞亚鹏; 刘宁; 汲恩民; 郝长虹; 赵广明; 吴建强
Original assignee: Hulunbuir Hengyi Agriculture And Animal Husbandry Co ltd
Current assignee: Hulunbuir Hengyi Agriculture And Animal Husbandry Co ltd
Priority date: 2018-02-05
Filing date: 2018-02-05
Publication date: 2022-07-08
Anticipated expiration: 2038-02-05
Also published as: CN110117552A

Abstract

The invention discloses a Bacillus subtilis HYNM-1 which is preserved in the China general microbiological center in 2017 in 10, 12 and the preservation number is CGMCC No. 14804. The strain has effect in inhibiting potato scab.

Description

Bacillus belgii and application thereof

Technical Field

The invention belongs to the technical field of bioengineering, and particularly relates to Bacillus velezensis and application thereof in preparation of a potato scab prevention and treatment microbial agent.

Background

The potato scab is a plant disease damaging potato tubers. The appearance of the disease is that the surface of the tuber is nearly round to irregular suberized scab-shaped light brown scab or plaque, and the hand feel is rough. Generally, two types of disease symptoms are developed, including a reticulated lesion and a lacerated lesion. Generally, although the disease spots are only limited to the skin layer, the quality and the yield of the damaged potato blocks can be reduced, the damaged potato blocks are not storage-resistant, the appearance of the damaged potato blocks is not elegant, the commodity grade is greatly reduced, and certain economic loss is caused. In China, the incidence of diseases is low, the area of damage is small, and therefore people do not pay attention to the disease. However, in recent years, the disease incidence area in inner Mongolia, Hebei, Shandong and other areas is increasing, and the economic loss caused by the disease incidence area is also considerable. Therefore, scientific control of such diseases is urgently required.

According to the long-term separation and identification research on pathogenic microorganisms, pathogenic bacteria causing potato scab are mainly streptomyces scabicus and the like. And the pathogenic bacteria belong to conditional pathogenic bacteria, namely the pathogenic bacteria easily grow and cause potato diseases under the alkaline condition of the soil environment, and rarely appear in the acidic soil environment.

The control method mainly comprises the following steps: (1) selecting seed potatoes without diseases; (2) applying organic fertilizer or green manure more times can partially inhibit diseases; (3) performing rotation with Cucurbitaceae, Leguminosae, and Liliaceae vegetables for more than 5 years; (4) selecting a vegetable field with good water retention for planting, and watering in time when meeting drought in a potato bearing period; (5) and chemical prevention: some chemical pesticides can be used for preventing and treating scab; (6) biological control method: the method screens microorganisms with good inhibition effect on scab pathogenic bacteria and strong planting capability, and is applied in the potato seed production process and the farmland planting process to achieve the effect of inhibiting the growth of the pathogenic bacteria.

Disclosure of Invention

The invention provides a Bacillus velezensis strain HYNM-1 which can effectively prevent and control potato scab.

The Bacillus subtilis HYNM-1 provided by the invention has been preserved in China general microbiological culture Collection center (CGMCC for short) in 10, 12 and 2017, and the preservation number is as follows: CGMCC No.14804, the preservation unit address is: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North.

Description of the drawings:

FIG. 1 is a graph showing the bacteriostatic effect of Bacillus belgii strain HYNM-1 on Streptomyces scabies 4.1765.

FIG. 2 is a graph showing the bacteriostatic effect of Bacillus belgii strain HYNM-1 on Streptomyces scabies 4.1765.

The specific implementation mode is as follows:

the strain is obtained by collecting soil sample from inner Mongolia potato scab disease soil, separating and identifying.

Colony morphology: the middle of a colony cultured on a potato culture medium for 2 days is raised and white, the surface wrinkles are irregular and dull, and the edge is irregular; the shape of the strain is: the strain is rod-shaped, has flagella, has a width of 0.76-0.94 μm and a length of 1.76-2.94 μm, and is aerobic, sporulating and gram-positive.

The identification method of the bacterial strain comprises the following steps: 16srRNA gene sequence analysis and identification as Bacillus velezensis.

The bacterial culture method comprises the following steps:

bacillus belgii was activated 3 times on potato solid medium. Then, 1 ring of the suspension was placed in 50ml of potato broth and cultured on a shaker at 28 ℃ for 2 days. After the fermentation broth was centrifuged at 12,000r/min for 1min, the supernatant was separately collected, and the precipitate was suspended in the same volume of sterile water to prepare a cell suspension. Meanwhile, the bacterial liquid and the supernatant are also used for the test.

The application and the effect of the strain are as follows:

the scab antagonistic bacteria screening material and method: the pathogen tested was Streptomyces scabies (Streptomyces scabeii) 4.1765 Potato juice medium (g/L): 1L of potato extract, 20g of glucose and KH₂PO₄ 3g，MgSO₄.7H₂O1.5 g and vitamin B1 in trace amount. pH6. solid medium 15g of agar was added.

The method for culturing pathogenic bacteria comprises the following steps: culturing pathogenic bacteria on potato culture medium for 4-5 days at 28 deg.C; scraping spores and mycelia with 0.2ml sterile water to obtain pathogen suspension with a certain concentration; the plate surface was covered with hyphae after growth when the plate was coated in a volume of 100 ul.

The bacteriostatic test method comprises the following steps: 100ul of pathogenic bacteria were spread on a Plate (PDA) and 10ul of bacterial suspension, supernatant, and cell suspension were spotted locally on the plate. Culturing at 28 ℃. Observed after 3 days. Through an experiment on Streptomyces scabies (Streptomyces scabeii) 4.1765, bacterial liquid and cell suspension can effectively inhibit the growth of pathogenic microorganisms, and supernatant has no inhibition effect.

As a result: the bacteria had an effect on pathogenic bacteria (fig. 1, fig. 2).

Claims

1. A Bacillus licheniformis (Bacillus siamensis) strain HYNM-1 with the preservation number of CGMCC No. 14804.

2. Use of a Bacillus belief (Bacillus siamensis) strain HYNM-1 according to claim 1 for the preparation of a medicament for inhibiting scab in potatoes.