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Method for quickly identifying plant endophytic bacteria producing auxin

A technology of plant endogenous bacteria and endogenous bacteria, which is applied in the field of microorganisms, can solve the problems of slow growth of bacteria, time-consuming, and time-consuming, and achieve the effects of effective identification, shortening time, and reducing experimental costs

Active Publication Date: 2015-09-02
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the process from reactivation, fermentation to final mixed color identification is extremely cumbersome, and it is necessary to replace the medium with different components many times and complicated sampling steps in the middle, and the growth of bacteria is slow in this process, which often consumes a lot of time (about 5~10 days)
Therefore, traditional screening methods for endophytic bacterial auxin-producing strains are tedious and time-consuming

Method used

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  • Method for quickly identifying plant endophytic bacteria producing auxin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] (1) Preparation of culture medium and chromogenic solution

[0024] Formulate modified R with 0.05% tryptophan 2 A separation and identification medium, the specific formula is as follows: Weigh 0.5 g yeast extract, 0.5 g tryptone, 0.5 g casein acid hydrolyzate, 0.5 g glucose, 0.5 g soluble starch, 0.3 g potassium dihydrogen phosphate, 0.024 g Magnesium sulfate water, 0.3 g sodium pyruvate, 15 g agar powder, 0.5 g tryptophan, dilute to 1 L with double distilled water, and adjust the pH to 7.2 to prepare the improved R 2 A Separation and identification medium, after autoclaving at 115 ℃ for 15 minutes, aliquoted and poured into 9 cm sterile petri dishes, cooled for later use.

[0025] Prepare auxin chromogenic solution: 2 mL of 0.5 M ferric chloride solution and 98 mL of 25% perchloric acid by volume and mix thoroughly.

[0026] (2) Pretreatment of plant tissue

[0027] Select well-growing plant tissues and rinse them with distilled water, then place them in an asepti...

Embodiment 2

[0032] (1) Preparation of culture medium and chromogenic solution

[0033] Formulate modified R with 0.1% tryptophan 2A separation and identification medium, the specific formula is as follows: Weigh 0.5 g yeast extract, 0.5 g tryptone, 0.5 g casein acid hydrolyzate, 0.5 g glucose, 0.5 g soluble starch, 0.3 g potassium dihydrogen phosphate, 0.024 g Magnesium sulfate water, 0.3 g sodium pyruvate, 15 g agar powder, 1 g tryptophan, dilute to 1 L with double distilled water, and adjust the pH to 7.2 to prepare the improved R 2 A Separation and identification medium, after autoclaving at 115 ℃ for 15 minutes, aliquoted and poured into 9 cm sterile petri dishes, cooled for later use.

[0034] Prepare auxin chromogenic solution: 2 mL of 0.5 M ferric chloride solution and 98 mL of 30% perchloric acid by volume and mix well.

[0035] (2) Pretreatment of plant tissue

[0036] Select well-grown plant tissues and rinse them with distilled water, then place them in an aseptic operating ...

Embodiment 3

[0041] (1) Preparation of culture medium and chromogenic solution

[0042] Formulate modified R with 0.2% tryptophan 2 A separation and identification medium, the specific formula is as follows: Weigh 0.5 g yeast extract, 0.5 g tryptone, 0.5 g casein acid hydrolyzate, 0.5 g glucose, 0.5 g soluble starch, 0.3 g potassium dihydrogen phosphate, 0.024 g Magnesium sulfate water, 0.3 g sodium pyruvate, 15 g agar powder, 2 g tryptophan, dilute to 1 L with double-distilled water, and adjust the pH to 7.2 to prepare the improved R2A separation and identification medium, after 115 ℃ After autoclaving for 15 min, aliquots were poured into 9 cm sterile Petri dishes, and cooled for later use.

[0043] Prepare auxin chromogenic solution: 2 mL of 0.5 M ferric chloride solution and 98 mL of 35% perchloric acid by volume and mix thoroughly.

[0044] (2) Pretreatment of plant tissue

[0045] Select well-grown plant tissues and rinse them with distilled water, then place them in an aseptic op...

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Abstract

The invention belongs to the technical field of microorganisms and relates to a method for quickly identifying plant endophytic bacteria producing auxin. The method comprises the following steps: (1) preparing a culture medium and a developing solution, wherein the used culture medium is an improved R2A isolation and identification culture medium which contains tryptophan with the concentration of 0.5-2g / L, and the developing solution of the auxin is obtained by uniformly mixing 2mL of a 0.5M ferric chloride solution with 98mL of perchloric acid of which the volume fraction is 25-35%; (2) pretreating plant tissues, namely soaking the plant tissues by using ethanol of which the volume fraction is 70-75% and a sodium hypochlorite solution of which the volume fraction is 5-8% respectively, and then flushing by using distilled water; and (3) identifying the endophytic bacteria producing the auxin. According to the method provided by the invention, an isolation culture medium and an identification culture medium producing the auxin are improved and integrated, precursor tryptophan for producing the auxin is added into the culture medium, the developing solution of the auxin is immediately applied after strain separation, and the endophytic bacteria producing the auxin is quickly identified in 5-10min; and the identification process is simplified, the operation is simple, and the time is shortened, so that the method can be used for simply, quickly and effectively identifying the endophytic bacteria producing the auxin.

Description

technical field [0001] The invention belongs to the technical field of microbes, in particular to a method for quickly identifying plant endophytic bacteria producing auxin. Background technique [0002] A large number of studies have proved that some plant endophytic bacteria can promote the absorption of nutrients by host plants and promote the growth and development of host plants and other physiological activities through biological nitrogen fixation and production of plant hormones ((1) Ansari, M. W., D. K. Trivedi, et al . (2013). A critical review on fungi mediated plant responses with special emphasis to Piriformospora indica on improved production and protection of crops. Plant Physiology and Biochemistry 70: 403-410.; (2) Rout, M. E. and T. H. Chrzanowski (2009) . The invasive Sorghum halepense harbors endophytic N2-fixing bacteria and alters soil biogeochemistry. Plant and Soil 315(1-2): 163-172.; (3) Rout, M. E., T. H. Chrzanowski, et al. (2013). Bacterial Endop...

Claims

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Application Information

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IPC IPC(8): C12Q1/04
Inventor 戴志聪付伟祁珊珊王晓莹蔡红红肖翔杜道林
Owner JIANGSU UNIV
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