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Preparation and application of a methylotrophic bacillus b18 and its liquid preparation

A methylotrophic and Bacillus technology, applied in the field of microorganisms, can solve the problems of ecological damage and easy pollution of the environment, and achieve the effects of low cost, avoiding environmental pollution, and good storage

Active Publication Date: 2020-06-09
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the prevention and control of diseases is mainly carried out through forest management measures to remove the source of disease, generally using chemical pesticides and chemical fertilizers. However, due to the long-term application of chemical pesticides (fertilizers) is easy to pollute the environment and damage the ecology, the discussion of comprehensive disease prevention and control measures has yet to be carried out.

Method used

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  • Preparation and application of a methylotrophic bacillus b18 and its liquid preparation
  • Preparation and application of a methylotrophic bacillus b18 and its liquid preparation
  • Preparation and application of a methylotrophic bacillus b18 and its liquid preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Isolation and Identification of Methylotrophic Bacillus B18

[0030] 1.1 Isolation, purification and preservation of endophytic bacteria

[0031] Healthy Phellodendron bark was collected from the area where Phellodendron rotten skin disease occurred in Yangtianwo Pharmacy, Dayi, Sichuan. Rinse the collected samples with sterile water, weigh 1.0g of tissue, soak in 70% alcohol for 1-2min, then disinfect with 1-3% sodium hypochlorite solution for 3-5min, wash several times with sterile water, absorb Apply 100 μL of washing liquid for the last time to NA plate (3g beef extract, 10g peptone, 5g sodium chloride, 15-20g agar powder, 1000mL distilled water, pH 7.0, after mixing and dispensing, autoclave at 121°C for 30min), culture in the dark at 27°C 24h, used as a control for surface disinfection, to check whether the surface disinfection is thorough.

[0032] Cut the surface-sterilized bark tissue into pieces, put them into a sterile mortar, add sterilized quart...

Embodiment 2

[0047] The preparation of embodiment 2 methylotrophic bacillus B18 liquid preparation

[0048] The methylotrophic bacillus B18 obtained according to Example 1 is made into a liquid preparation through seed culture and fermentation culture, specifically through the following methods:

[0049] The conventional method is to prepare beef extract peptone slant medium (beef extract 7g, peptone 10g, NaCl 5g, agar 18g, water 1000mL), inoculate methylotrophic Bacillus, and cultivate it to make first-class seeds.

[0050] Nutritious succulent culture solution (beef extract 7g, peptone 10g, NaCl 5g, Phellodendron twig impregnation solution 1000mL) is autoclaved, and under the control of oxygen, it is bottled according to the ratio of 100mL liquid to 300mL triangular flask. Inoculate methylotrophic bacillus slant seeds, inoculate 2 slant seeds in each bottle, vibrate culture, and make methylotrophic bacillus liquid seeds. Wherein 10g of Phellodendron twigs was boiled in 1000mL water for ...

Embodiment 3

[0055] Embodiment 3 potted plant control effect test

[0056] Select 60 annual potted Phellodendron Phellodendron seedlings as the experimental objects, and conidia of Nectriahaematococca (spore concentration: 1×10 6 cfu / mL) 30mL after acupuncture inoculation stem 15d, adopt spraying method to divide different concentrations to treat Cortex Phellodendri seedlings with the bacterial agent that embodiment 2 makes, every pot 100mL, every 7 sprays 1 time (totally 3 times), And use sterile water as a control, repeat 3 times, and count the incidence of Cork Phellodendron. Treat 10 Phellodendron seedlings per treatment.

[0057] After 30 days of inoculation with pathogenic bacteria, conduct disease investigation and statistics according to the disease grading standards in Table 3, and calculate the incidence rate, disease index and control effect.

[0058] Table 3 Grading standards of Phellodendron cork rot disease

[0059]

[0060] Incidence rate (%)=number of diseased plants / ...

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Abstract

The invention discloses the preparation and application of a methylotrophic Bacillus B18 and its liquid preparation. The strain has been preserved in the China General Microorganism Culture Collection Management Center on September 18, 2017, and the preservation number is CGMCC No.14641 . The liquid preparation can be prepared by using the methylotrophic bacillus B18 as the strain, culturing seeds and fermenting. The methylotrophic Bacillus B18 provided by the invention has special effects on Phellodendron rot, and has stress resistance, good storage performance, high temperature resistance, low temperature resistance, and drying resistance, is suitable for commercial production, and can not only avoid chemical drugs. The serious consequences of polluting the environment, and it is sustainable and low-cost.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a methylotrophic bacillus B18 strain and its liquid preparation and its application in the prevention and treatment of cork rot. Background technique [0002] Phellodendron Phellodendri is the main raw material for extracting berberine. It is also an important economic tree species and traditional Chinese medicinal material for skin in my country. It has a medicinal history of more than 2,000 years. The bark contains a variety of alkaloids, which have a special aroma and bitter taste. It has the functions of removing steam by fire, purging fire and detoxification, reducing swelling and removing rot, and is a valuable medicinal resource. Phellodendron cortex is a commonly used traditional Chinese medicine for clearing away heat and dampness. It has a wide range of uses and a large market demand. It can be made into a variety of Chinese patent medicines, cosmetics...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P3/00C12R1/07
CPCA01N63/10C12N1/20C12N1/205C12R2001/07
Inventor 李姝江朱天辉方馨玫何倩倩黎肇家谯天敏韩珊
Owner SICHUAN AGRI UNIV
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