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Bacillus amyloliquefaciens jk6 and biological fertilizer and its application

A technology of amylolytic spores and JK6, which is applied in the direction of biochemical equipment and methods, applications, microorganisms, etc., and can solve the problems of quantitative detection of functional gene expression in soil and the lack of in-depth research on bacterial strains, etc.

Active Publication Date: 2018-04-06
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although many biocontrol strains have been reported to contain a variety of functional genes for disease prevention, whether the strains can successfully colonize and express in large quantities in complex soil environmental factors has not been studied in depth, and the quantitative detection method for the expression of functional genes in soil remains to be done. Research

Method used

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  • Bacillus amyloliquefaciens jk6 and biological fertilizer and its application
  • Bacillus amyloliquefaciens jk6 and biological fertilizer and its application
  • Bacillus amyloliquefaciens jk6 and biological fertilizer and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] Example 1 Isolation and purification of bacterial strains

[0094] (1) Separation

[0095]Beef extract-peptone medium: 10g bacteriological peptone, 3g beef extract, 5g sodium chloride, 18g agar, 1000mL distilled water, adjust the pH to 7.0-7.2.

[0096] Preparation of plates containing Ralstonia solanacearum: make bacterial suspension of Ralstonia solanacearum, use a pipette gun to draw 5mL of Ralstonia solanacearum suspension and add it to 100mL of beef that has been heated and melted and the temperature has dropped to about 50°C In the peptone medium, shake well, pour the plate, and let it dry naturally to make a plate containing R. solanacearum for later use.

[0097] Take the red soil in the tomato planting base of the South China Agricultural University farm in Guangzhou as the screening soil sample, accurately weigh 10 g of the screening soil sample by the flat plate coating method, put it into a 250 mL triangular flask with 90 mL of sterile water (with glass bea...

Embodiment 2

[0100] Example 2 Characterization

[0101] (1) Morphological characteristics of bacteria

[0102] Gram-positive bacteria with spores. The bacteria are rod-shaped, motile and aerobic. Gram-stained micrographs of bacteria figure 1 , see photos of spore staining figure 2 .

[0103] (2) Colony Morphological Characteristics

[0104] After 24 hours of cultivation on the beef extract peptone medium, the colonies formed were round or irregular. After 48 hours, the colonies were all round, milky white, about 1-2 mm in diameter, with irregular edges, flat and wet, and biofilms formed on the surface.

[0105] (3) Growth characteristics

[0106] In the liquid medium of 5g of yeast extract powder, 10g of bacteriological peptone, 10g of sodium chloride, and 1000mL of water, the rotation speed is 150rpm / min, the temperature is 30°C, and the initial pH value is 7.0, cultured for 18h, and the viable bacteria were measured The number is (3.95±0.60)×10 8 cfu / mL.

[0107] (4) Physiologi...

Embodiment 3

[0115] Embodiment 3 JK6 bacterium produces catalase test

[0116] After activating the JK6 bacteria on the beef extract peptone medium for 24 hours, pick an appropriate amount of the test strain on the glass slide, drop 3% hydrogen peroxide on the test strain, observe immediately, if there are a lot of bubbles, it is positive, otherwise is negative. Taking HL-1 as the positive control, the test results are as follows: Figure 4 As shown, it shows that the catalase test of JK6 bacteria is positive.

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Abstract

The invention discloses bacillus amyloliquefaciens JK6 and biological fertilizer and application. The serial number of strain preservation is CGMCC No. 10658 and is stored in the China general microbiological culture collection center on March 23, 2015. The bacillus amyloliquefaciens JK6 is mainly used for biological prevention and control of pseudomonas solanacearum and is provided with an iron-production carrier, a biological membrane, protease, catalase, cellulose and IAA characteristics, multiple fungal pathogens of banana wilt pathogen, litchi anthrax germs, litchi frost phytophthora pathogens, rice blast fungus, cucumber wilt pathogen and the like, and growth of lettuce plants is obviously promoted; the K6 strain can be successfully planted, and disease preventing functional genes of yndj, srfAB, fend and ituC are expressed in soil; the strain is high in reproduction speed, simple in production technology, high in adverse resistance, easy to store and favorable for industrialized production, the biological fertilizer is prepared with the strain, soil-born diseases can be prevented and controlled, growth of the plants can be promoted, and application prospect is broad.

Description

technical field [0001] The invention relates to a microorganism and an application thereof, in particular to a Bacillus amyloliquefaciens JK6 (Bacillus amyloliquefaciens) and an application thereof. Background technique [0002] Tomato bacterial wilt is a destructive soil-borne disease caused by Ralstonia solanacearum, and the incidence rate in severely diseased fields can be as high as 80%, or even result in crop failure. At present, the control of tomato bacterial wilt in agriculture mainly relies on chemical pesticides, and the effect is not ideal. Long-term large-scale and irrational application of chemical fertilizers and chemical pesticides will easily lead to drug resistance of pathogenic bacteria and increase of toxic chemical residues in fruits and vegetables. With the development of green agriculture, it is gradually turning to comprehensive control based on biological control. Screening highly effective antagonistic strains is the core of biological control. Stud...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N15/31C12Q1/689C05F11/08A01N63/00A01P3/00A01P1/00A01P21/00C12R1/07
Inventor 蔡燕飞熊汉琴李永涛郭真真张娅娅江亚雄
Owner SOUTH CHINA AGRI UNIV
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