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Acquisition and application of a ginseng growth-promoting bacterium producing acc deaminase

A ginseng endophytic bacteria and ginseng technology, applied in the field of microorganisms, can solve the problems of slow growth, low germination rate of ginseng seeds, poor stress resistance, etc., and achieve the effect of promoting growth

Inactive Publication Date: 2016-06-08
JILIN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The object of the present invention is to provide a ginseng growth-promoting bacterium that produces ACC deaminase for overcoming the shortcomings of ginseng seeds such as low germination rate, slow growth, and poor stress resistance

Method used

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  • Acquisition and application of a ginseng growth-promoting bacterium producing acc deaminase
  • Acquisition and application of a ginseng growth-promoting bacterium producing acc deaminase
  • Acquisition and application of a ginseng growth-promoting bacterium producing acc deaminase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Obtaining endophytic bacteria (Pseudomonas fluorescens) producing ACC deaminase activity of ginseng.

[0020] 120 strains of ginseng endophytic bacteria from Jilin Province were isolated by plate dilution method, and the strains with ACC deaminase activity were screened.

[0021] The screening is divided into primary screening and re-screening. The liquid culture method is used to screen 120 strains of bacteria in the primary screening. The strains to be tested are respectively inoculated into liquid LB medium and cultivated overnight at 28°C and 160r / min. Transfer 0.2 mL of culture medium to 8 mL of DF salt medium, and culture for 12 hours under the same culture conditions. Then take 0.2mL medium and transfer to 8mL ADF liquid medium again. And the ADF medium without ACC was used as the control. After culturing for 24-48 hours, measure the absorbance value at 600nm, repeat three times, and the positive strain is the one whose absorbance value is significant...

Embodiment 2

[0023] Example 2: Determination of other growth-promoting indicators of active bacterial strain JJ8-3

[0024] 1. Screening for the ability to decompose inorganic phosphorus: The PKO solid culture method was used for the primary screening. After culturing the JJ8-3 strain on NA medium for 12 hours, inoculate the bacteria plate on the PKO solid medium, and observe the phosphorus solubilization after culturing at a constant temperature of 28°C for 3 days. circle size. The second screening was carried out by molybdenum-antimony-scandium colorimetric method. JJ8-3 was inoculated into 50mL LB liquid medium by punching holes, and cultured at 160r / min for 12h. Take 1mL and inoculate it in a 150mL Erlenmeyer flask containing 50mL of PKO liquid medium. After 3 days, take 5mL of the culture solution, centrifuge at 8000r / min for 5min, and take the supernatant. Measure the soluble phosphorus content by molybdenum-antimony-scandium colorimetry. The results showed that on the PKO plate, th...

Embodiment 3

[0027] Embodiment 3: Identification of bacterial strain JJ8-3

[0028] Streak inoculation of JJ8-3 strain on NA medium, culture at 28°C for 12-24 hours, observe and record single colony morphology. JJ8-3 showed a light green opaque circular colony on NA medium, with complete and slightly transparent edges ( image 3 ), negative Gram staining, no motility, fluorescence observed under ultraviolet light ( Figure 4 ).

[0029]Take the JJ8-3 strain cultured in LB liquid medium for 24 hours, centrifuge and wash repeatedly with sterile water, suspend in an appropriate amount of sterile water for negative staining to prepare samples, and use tweezers to clamp the copper mesh with film to dip a small amount Bacteria solution, let it stand for 2min, blot the remaining liquid from the edge with filter paper; then use tweezers to clamp the copper mesh with bacteria and immerse it in a beaker filled with double distilled water for rinsing, then use filter paper to blot the liquid, and t...

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Abstract

The invention discloses JJ8-3 bacterial strains of pseudomonas fluorescens of ginseng endophytic bacteria, and the preservation number is CGMCC No.8239. Among plenty of ginseng endophytic bacteria, some bacterial strains which can produce ACC deaminizing enzyme are obtained through primary selection, and the bacteria for producing the ACC deaminizing enzyme and promoting the growth of ginseng are obtained through repeatedly screening. Plenty of experiments prove that the bacterial strains JJ8-3 can produce the ACC deaminizing enzyme and have strong action for promoting the growth of the ginseng at the same time, and the bacterial strains have no pathogenicity. Through identification, the bacterial strains JJ8-3 are pseudomonas fluorescens. The disadvantages of low germination rate of ginseng seeds, slow growth of the ginseng which is one of the most important traditional Chinese medicine materials in China, poor stress resistance of the ginseng and the like are solved. The finding of the bacterial strains establishes a favorable basis for promoting the growth of the ginseng and developing and utilizing bacterial manure.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, in particular to a ginseng growth-promoting bacterium producing ACC deaminase and its application. Background technique [0002] In recent years, the occurrence of natural disasters and the irrational use of chemical fertilizers have had a great negative impact on soil and crop growth. It is of great significance to screen microbial strains from the environment that can promote plant growth, improve plant stress resistance and improve the environment. Some strains with growth-promoting functions have been reported so far, and their main characteristics include IAA production, nitrogen fixation, phosphorus and potassium dissolution, and siderophore production. However, there are few reports on 1-aminocyclopropane 1-carboxylic acid (ACC) deaminase and its active strains, and it is also one of the characteristics of plant growth-promoting bacteria that has attracted more attention in recent ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/02A01P21/00C12R1/39
Inventor 陈长卿姜云田磊张冠军李桐
Owner JILIN AGRICULTURAL UNIV
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