A rapid method for the identification of auxin-producing endophytic bacteria

A technology of plant endogenous bacteria and endogenous bacteria, which is applied in the field of microbiology, can solve the problems of cumbersome process, time-consuming, slow growth of bacteria, etc., and achieve the effect of simplifying the identification process, reducing the cost of experiments, and effectively identifying

Active Publication Date: 2017-10-20
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • A rapid method for the identification of auxin-producing endophytic bacteria

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Experimental program
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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 well.

[0026] (2) Pretreatment of plant tissue

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

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 thoroughly.

[0035] (2) Pretreatment of plant tissue

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

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 microbes, and relates to a method for quickly identifying endophytic bacteria producing auxin; it includes the following steps: (1) preparation of a culture medium and a chromogenic solution: the culture medium used contains tryptophan at a concentration of 0.5 ‑2 g / L improved R2A isolation and identification medium, the auxin chromogenic solution is 2 mL of 0.5 M ferric chloride solution and 98 mL of perchloric acid with a volume fraction of 25-35%; (2) Pretreatment of plant tissue: after soaking in ethanol with a volume fraction of 70-75% and sodium hypochlorite solution with a volume fraction of 5-8%, rinse with distilled water; (3) identification of endophytic bacteria producing auxin; Integrate with the improvement of auxin-producing identification medium, add auxin-producing precursor tryptophan to the medium, apply auxin chromogenic solution immediately after the strain is isolated, and quickly identify the auxin-producing auxin within 5-10 minutes Endophytic bacteria: Simplified identification process, convenient operation, shortened time, is a simple, rapid and effective method for identifying auxin-producing endophytic bacteria.

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). invasive Sorghumhalepense harbors endophytic N2-fixing bacteria and alters soilbiogeochemistry. Plant and Soil 315(1-2): 163-172.; (3) Rout, M. E., T. H. Chrzanowski, et al. (2013). . American Journal of Bo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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