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Acid-resistant bacillus amyloliquefaciens Kc-5 and application thereof

A technology of amylolytic spores and kc-5, which is applied in the field of microorganisms to achieve strong antibacterial activity, good disease prevention and growth promotion effects, and a wide antibacterial spectrum

Active Publication Date: 2019-11-15
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the use of acid-resistant bacillus and microbial preparations, microbial fertilizers or soil improvers prepared from it for plant disease biocontrol and soil improvement has not been reported.

Method used

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  • Acid-resistant bacillus amyloliquefaciens Kc-5 and application thereof
  • Acid-resistant bacillus amyloliquefaciens Kc-5 and application thereof
  • Acid-resistant bacillus amyloliquefaciens Kc-5 and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Isolation, identification and biological characteristics of strains

[0020] 1. Isolation and purification of strains

[0021] In this experiment, several strains of bacteria were isolated from the acidic soil in Guangxi. The process was as follows: 1 g of the sample was added to a conical flask filled with small glass beads and 100 ml of sterile saline, enriched and cultured on a shaker at 28°C for about 30 minutes. Take 1m suspension and dilute to 10 by gradient dilution method -4 , take stock solution, 10 -2 Diluent and 10 -4 The dilutions were plated separately and cultured for at least two weeks. After the grown colonies were purified on the LB plate by streaking the plate method, single colonies were picked and inoculated on the LB plate. The resulting purified strains were stored in a -80°C refrigerator using 20% ​​glycerol.

[0022] 2. Identification of strains

[0023] 1. Molecular identification

[0024] 16S rDNA was amplified by PCR using universal pri...

Embodiment 2

[0040] Antifungal activity analysis of acid-resistant Bacillus amyloliquefaciens Kc-5

[0041] 1. Culture and collection of Bacillus amyloliquefaciens Kc-5

[0042]Inoculate Kc-5 on the LB plate by streaking method, and after a single colony grows, pick a single colony and inoculate it in liquid LB medium for 24 hours at 30°C before use.

[0043] 2. The cultivation of pathogenic bacteria

[0044] Inoculate banana Fusarium wilt, cucumber Fusarium wilt, wheat root rot umbilicaleum, capsicum phytophthora, and vegetable gray mold on PDA plates, culture at 28°C for one week before use.

[0045] 3. Tablet confrontation experiment results

[0046] Inoculate the bacterial block of pathogenic bacteria culture in the center of the PDA plate, inoculate the suspension of Bacillus amyloliquefaciens Kc-5 about 2.0cm away from the pathogenic bacteria, culture at 28°C for 3-5 days, and observe the growth of the pathogenic bacteria.

[0047] Acidfast Bacillus amyloliquefaciens Kc-5 isolated...

Embodiment 3

[0050] Preparation of Acid-resistant Bacillus amyloliquefaciens Kc-5 Inoculant

[0051] 1. Preparation of Seed Solution

[0052] Pick a single colony of Kc-5 grown on the LB plate for 24 hours, inoculate it into a 500ml Erlenmeyer flask containing 150ml liquid LB medium, and culture it at 37°C and 180rpm for 23 hours. Then transfer to a new 1 Erlenmeyer flask containing 150ml of liquid LB medium according to the ratio of 3-5%, shake and cultivate for 6 hours, and use it as a seed solution.

[0053] 2. Preparation of liquid fermentation microbial inoculum

[0054] 50L fermenter contains 30L medium: yeast powder 3g / l, peptone 5g / L, beef extract 2g / L, sodium chloride 2g / L, MnSO 4 0.005g / L, K 2 HPO 4 3g / L, MgSO 4 0.02g / L. pH: keep the pH at 7.0, and adjust it with ammonia water during the fermentation process.

[0055] DO: Dissolved oxygen is related to the speed, keep around 30%.

[0056] Tank pressure: 0.04Mpa.

[0057] Glucose feeding: start feeding when the residua...

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Abstract

The invention provides acid-resistant bacillus amyloliquefaciens Kc-5 and application thereof. The bnacillus amyloliquefaciens Kc-5 is preserved in the China General Microbiological Culture CollectionCenter, CGMCC for short, on April 29th, 2019, the address is Building No.3, NO.1 West Beichen Road, Chaoyang District, Beijing, and the preservation number of the bnacillus amyloliquefaciens Kc-5 isCGMCC No.17654. The nacillus amyloliquefaciens Kc-5 has a wide pH suitable range of 4-10 and good acid resistance; the strain has a broad antibacterial spectrum and has a good antagonism effect on various plant pathogenic fungi such as fusarium oxysporum, fusarium oxysporun f.sp.cubense, phytophthora capsici, botrytis cinerea and bipolaris sorokinianum, and it is shown through potting and field tests that the strain has good disease prevention and growth inhibition effects on the fusarium oxysporun f.sp.cubense in acid soil. The strain is high in adaptability and antibacterial activity and canbe used in neutral and acid soil for efficiently preventing and controlling fungal diseases; the strain is also suitable for improving the acid soil.

Description

technical field [0001] The invention relates to the technical field of microorganisms. Background technique [0002] Fungal diseases account for more than 70% of plant invasive diseases, and are the main reason for the reduction of yield and quality of crops and fruit trees. The frequent use of chemical pesticides has led to a sharp increase in the resistance of some pathogenic fungi, pollution of the environment and agricultural products, and damage to the balance of microorganisms in and out of plants, which has exacerbated the outbreak of diseases (Yu Fengyu et al. 2005). The use of microorganisms for the biological control of fungal pathogens is an ideal alternative. Biological control has no residue and no pollution, no resistance to pathogenic bacteria, low cost, increased production and income, is conducive to the sustainable development of agriculture, and has broad prospects. [0003] Bacillus is one of the biocontrol bacteria with the most potential and commercia...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05G3/00A01N63/00A01P3/00A01P21/00C12R1/07
CPCA01N63/00C05G3/00C12N1/205C12R2001/07
Inventor 孙宪昀李少杰张振颖
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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