Chitosanase-producing and fungus-restraining Burkholderia sp. and application thereof

A technology for producing chitosan enzymes and fungi, applied in the field of microorganisms, to achieve significant growth-promoting effects, wide application prospects, and increased plant height

Active Publication Date: 2016-10-12
东辽县日月星有机肥有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are few reports on Burkholderia, which can both inhibit plant pathogenic fungi and improve soil fertilizer efficiency

Method used

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  • Chitosanase-producing and fungus-restraining Burkholderia sp. and application thereof
  • Chitosanase-producing and fungus-restraining Burkholderia sp. and application thereof
  • Chitosanase-producing and fungus-restraining Burkholderia sp. and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The screening of embodiment 1 bacterial strain of the present invention

[0031] Water samples and soil collected near Changqing Garden Park in Wuhan were added to sterilized water and shaken vigorously. Sample: sterile water=1:9 (the water sample is the volume ratio, and the soil sample is the mass-to-volume ratio), and the resulting suspension concentration is 10 -1 , and then the above concentration is 10 -1 The suspension was diluted to 10 -2 ~10 -6series concentration. Take 200 μL of the above-mentioned dilutions of different concentrations and evenly spread them on a chitosan screening plate, incubate at a constant temperature for 3 to 5 days by inverting at 28°C. Pick colonies that grow well and can produce transparent circles for streak purification, and use glycerol preservation method to preserve bacteria.

[0032] Through the above screening, a strain that can use chitosan as the sole carbon source to produce obvious hydrolysis transparent circles on the...

Embodiment 2

[0033] Morphological observation and molecular biological identification of the strain of embodiment 2

[0034] (1) Morphological and physiological and biochemical characteristics

[0035] Morphological, physiological and biochemical characteristics of the strain MEL01: cultured on a beef extract peptone plate at 28°C for 48 hours, the colony is round, with neat edges, light yellow opaque, moist and viscous surface, initially flat, and grows with the growth of culture time The colonies become thicker, see the specific morphology figure 2 . According to "Bergey's Handbook of Bacteria", MEL01 also has the following physiological and biochemical properties: negative Gram staining, short rod-shaped bacteria, no spores, negative denitrification, positive citrate utilization, positive leucine utilization, catalase Positive, gelatin liquefaction positive.

[0036] (2) 16S rDNA molecular identification

[0037] Preparation of PCR amplification template: Pick a single colony of ME...

Embodiment 3

[0041] Embodiment 3 bacterial strain degrades chitosan performance measurement

[0042] (1) Qualitative experiment of degrading chitosan transparent circle method

[0043] Take 10 μL of the MEL01 bacterial suspension activated by beef extract peptone liquid medium for 10 hours and plant it on a chitosan plate, plant 4 strains symmetrically at the four corners of the plate, and culture it upside down in a 37°C incubator for 3 days, MEL01 can form a very obvious hydrolysis circle, see the result Figure 4 . Use a vernier caliper with an accuracy of 0.02mm to measure the ratio of the diameter (D) of the hydrolysis zone of each colony to the diameter (d) of the colony, and the D / d is more than 3.5 (the larger the D / d value, the stronger the degradation ability), it can be considered that MEL01 has a significant The degradation effect, the specific measurement results are shown in Table 1.

[0044] (D / d) ratio of chitosan degraded by strain MEL01 in table 1

[0045]

[0046]...

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Abstract

The invention discloses a Burkholderia sp. and application thereof, and belongs to the field of microorganisms. The Burkholderia bacterium Burkholderia sp. MEL01 provided by the invention had been collected in the China Center for Type Culture Collection with a collection number of CCTCC NO.M2015778 and on a collection date of December 25, 2015. The bacterial strain has higher chitosanase activity, and further has obvious antagonistic actions on six vegetal pathogenic fungi of a strawberry gray mold and the like; by utilizing a characteristic that the bacterial strain MEL01 can degrade chitosan, the MEL01 is mixed with colloidal chitosan to make the obtained mixture into composite bacterial fertilizer which can obviously improve the soil fertility of a crop; the Burkholderia sp. can be applied to the biological prevention and control of a vegetal pathogenic bacterium and the production of novel biological fertilizer, and has a wide application prospect in an aspect of ecological agriculture.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a strain of Burkholderia producing chitosanase and inhibiting fungi and its application. Background technique [0002] Chitin is widely found in the cell walls of fungi and algae in nature, the outer shells of arthropods, and the inner shells and cartilages of molluscs. Chitosan is the product of deacetylation of chitin, and chitin with a deacetylation degree of more than 70% is usually called chitosan. The molecular formula of chitosan is (C 6 h 11 NO 4 )n, has the characteristics of degradability, adsorption, moisture retention, biocompatibility, etc., and can be widely used in the fields of industry, agriculture, medicine, environmental protection and health. However, chitosan itself is insoluble in water and degrades slowly, making it difficult to be absorbed and utilized by organisms. Therefore, how to degrade chitosan into chitooligosaccharides with ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/04A01P3/00C05F15/00C12R1/645
CPCA01N63/30C05F11/00C05F11/08C12N1/145C12R2001/645
Inventor 倪红唐梦君邵闯万娟刘诚刘源彬
Owner 东辽县日月星有机肥有限公司
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