Biocontrol bacterium I-5 and application thereof in prevention and treatment of Medicago sativa root rot

A technology of alfalfa and bio-control bacteria, applied in the biological field, can solve the problems of killing non-pathogenic beneficial microorganisms, poor environment and human health, and drug resistance of bacteria, achieving good thermal stability, good stability, Effects of Soil Invertase and Soil Neutral Phosphatase Activity Promotion

Active Publication Date: 2020-11-10
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, my country mainly relies on agricultural control and chemical control to control root rot, but the soil environment and farming system in different planting areas are different, and the extent of relying on agricultural control to control the disease is limited. Although chemical control measures are effective, they are soil-borne. For diseases, it is difficult to apply after the onset of the disease, and the large amount of medication is not effective for the environment and human health, and it will kill non-pathogenic beneficial microorganisms, and it will also make the bacteria resistant to antibiotics

Method used

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  • Biocontrol bacterium I-5 and application thereof in prevention and treatment of Medicago sativa root rot
  • Biocontrol bacterium I-5 and application thereof in prevention and treatment of Medicago sativa root rot
  • Biocontrol bacterium I-5 and application thereof in prevention and treatment of Medicago sativa root rot

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Isolation and screening of alfalfa root rot biocontrol bacteria

[0045] (1) Separation of biocontrol bacteria

[0046] Use the dilution method to separate from the rhizosphere soil of alfalfa that has been continuously cropped for 13 years. The specific operation is as follows: take 10g of rhizosphere soil samples and place them in a triangular flask (500mL) filled with 200mL of sterile water, and shake for 20min at 28°C and 170rpm Afterwards, take 1 mL of the supernatant and add it into a test tube filled with 9 mL of sterile water, and configure a concentration gradient of 10 -1 g / mL, 10 -2 g / mL, 10 -3 g / mL, take 0.5mL solution in a test tube and inject it into a sterile petri dish, add beef extract medium (15mL) into the petri dish, mix well, dry it, seal it with a parafilm and place it at a constant temperature of 26°C for incubation Cultivate in the box for 2-3 days, and when the single colony grows on the surface of the medium, pick it out with a tra...

Embodiment 2

[0060] Example 2 Identification of biocontrol bacteria for alfalfa root rot

[0061] According to "Berger's Bacteria Identification Handbook", "Common Bacteria System Identification Handbook" edited by Dong Xiuzhu and Cai Miaoying and other identification standards, the morphological characteristics and physiological and biochemical characteristics of the biocontrol strain were identified.

[0062] (1) Morphological identification

[0063] A. Morphological characteristics: Pick a small amount of target biocontrol bacteria I-5, inoculate it in NA liquid medium, place it under the condition of 28°C and 170rpm for 24 hours to obtain a bacterial suspension, and dilute it by 10 6 After doubling, take 15 μL of the bacterial suspension, spread it evenly on the NA medium plate, and incubate it in a 28°C incubator at a constant temperature for 2-3 days until a single colony appears, then observe its morphological characteristics.

[0064] B. Gram staining: drop distilled water on the ...

Embodiment 3

[0094] Example 3 Biocontrol bacterium I-5 measures the control effect of indoor pot test on alfalfa root rot

[0095] Preparation of bacterial suspension: put the antagonistic target biocontrol bacteria I-5 on the NA medium plate, place it in a constant temperature incubator at 28°C for 4-5 days, and pour it into the cultured NA plate 0.9% sterile saline, gently scrape the bacteria on the surface of the culture medium with a transfer hook, and measure the OD under the spectrophotometer 600 The value is 0.1 (equivalent to a cell concentration of 1×10 8 cfu / mL).

[0096] Preparation of spore suspension: Inoculate Fusarium tritinabilis of alfalfa root rot on a PDA medium plate, place it in an incubator at 28°C for propagation and culture for 4-5 days, add 15 mL of sterile water dropwise to the petri dish, and transfer bacteria with Scrape the mycelium of pathogenic bacteria on the surface of the PDA medium lightly with a hook, filter it through double-layer gauze, place it unde...

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Abstract

The invention discloses a biocontrol bacterium I-5 and application thereof in prevention and treatment of Medicago sativa root rot, and belongs to the technical field of biology. According to the invention, Fusarium tricinctum is used as a target pathogenic bacterium, biocontrol bacteria separated from Medicago sativa rhizosphere soil are screened by using a confrontation culture method to obtaina target biocontrol bacterium I-5, and the target biocontrol bacterium I-5 is determined as ochrobactrum intermedium. The biocontrol bacterium I-5 sterile filtrate plays a good role in inhibiting thegermination and production of Fusarium tricinctum spores of root rot bacteria and the growth of hyphae; the bacteriostatic active substance shows good thermal stability, is relatively sensitive to acid-base change, and has good stability to ultraviolet rays; the biocontrol bacterium I-5 has an obvious promoting effect on the activity of soil sucrase and soil neutral phosphatase, has a small increase in the activity of soil cellulase and urease, and has no obvious promoting effect on the activity of soil catalase; and the biocontrol bacterium I-5 can obviously promote the fresh weight and plantheight of Medicago sativa, and has no phytotoxicity.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to a biocontrol fungus I-5 and its application in preventing and treating alfalfa root rot. Background technique [0002] Alfalfa (Medicago sativa L.) is the leguminous forage with the largest planting area in the world, and root rot is one of the main diseases that cause alfalfa diseases. In the region, there are many pathogenic factors that cause root rot, and the disease can cause severe occurrence in a large area, resulting in the death of a large area of ​​plants, which will have a greater impact on the quality and yield of alfalfa, causing economic losses. If alfalfa disease occurs in the roots, the population density will decrease continuously, and the number of plants will drop to 16% of the original unit area within a few years, and the sustainable use value of grassland will be completely lost. In recent years, the planting area of ​​alfalfa has continued to e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/20A01P3/00A01P21/00C12R1/01
CPCC12N1/20A01N63/20C12R2001/01C12N1/205Y02P60/21
Inventor 李永刚崔国文陈雅君姜婉怡刘金鑫蔡亚男
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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