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Salt-tolerant bacteria T5S2 and applications in microbial fertilizer

A microbial fertilizer, T5S2 technology, applied in the direction of microorganism-based methods, microorganisms, microorganisms, etc., can solve the problem of halomonas rare and other problems, and achieve the effects of obvious salt reduction and production increase, fast reproduction, and various application methods

Active Publication Date: 2016-11-09
新疆山川秀丽生物有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing technology studies have shown that the research on Halomonas in biological control and microbial fertilizers is relatively rare. Therefore, it is of practical significance to study the characteristics and applications of new strains of extremophiles

Method used

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  • Salt-tolerant bacteria T5S2 and applications in microbial fertilizer
  • Salt-tolerant bacteria T5S2 and applications in microbial fertilizer
  • Salt-tolerant bacteria T5S2 and applications in microbial fertilizer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Isolation, screening and identification of Halomonas sp. T5S2

[0033] 1. Isolation: It was isolated from rhizosphere soil samples of Yanzhaozhao in the saline-alkali area of ​​Junggar Basin, Xinjiang. Weigh 1g of soil sample and suspend it in the Erlenmeyer flask containing PAF culture medium, shake and cultivate at 30°C and 150r / min for 24h, draw 1ml of bacterial suspension into 50ml of DF culture medium, shake and cultivate at 30°C and 150r / min for 24h, use inoculation The loop inoculation bacteria suspension was placed in ADF culture medium, 30°C, 150r / min shaking culture for 24h, after repeated transfer twice, gradient dilution, spread on ADF solid plate, 28°C constant temperature culture, until the plate grows a single colony.

[0034] 2. Purification: After the bacterial colonies grow, pick colonies of different shapes, sizes, colors, etc., and inoculate them on corresponding plates, and purify and cultivate them at 30°C-37°C, preferably at 33°C until ...

Embodiment 2

[0044]Example 2: Halomonas sp. T5S2CGMCC No.12642 produces ACC deaminase activity in different media

[0045] The strain T5S2 was inoculated respectively in LB, TSB and ADF culture fluid containing 1% NaCl, 30°C, 150rpm shaking culture for 2 days, then centrifuged at 4°C to collect the bacteria, and washed with 0.1mol / L Tris-HCl buffer (pH=7.6 ) after washing for 3 times, resuspend in 600 μl 0.1 mol / L Tris-HCl buffer (pH=8.0), add 30 μl toluene and shake rapidly for 30 s to break the cells. Take 200 μL of the above cell extract, add 20 μl 0.5mol / L ACC and mix well, and do a blank test without ACC at the same time, in a constant temperature water bath at 30°C for 15 minutes. Add 1ml of 0.56mol / L HCl, centrifuge at 16,000g for 5min, take 1ml of this suspension, add 800μl of 0.56mol / L HCl and 300μl of 2,4-dinitrophenylhydrazine, put in a constant temperature water bath at 30°C for 30min, add 2ml of 2mol / L NaOH, 540nm Measure the absorbance.

[0046] See the attached figure 2 ...

Embodiment 3

[0047] Example 3: Halomonas sp. T5S2CGMCC No.12642 produces IAA in different culture media

[0048] Bacterial strain T5S2 was respectively inoculated in LB, TSB and indole acetic acid (lAA) medium containing 1% NaCl, 30 ° C, 150 rpm shaking culture for 2 days, centrifuged to get 1 ml of supernatant, added 50 μl of 10 mmol / L orthophosphoric acid, Add 2ml of Salkowski's chromogen, develop color at 25°C for 30min in the dark, and measure the absorbance at 530nm wavelength. Using distilled water instead of the culture medium, the same reaction was used as a control to adjust to zero, and different concentrations of IAA standard solutions were used to make a standard curve in the same way, and the concentration of IAA in the fermentation broth was calculated.

[0049] See the attached image 3 , the strain T5S2 can produce IAA in the three mediums of the experiment, and the yield of IAA-producing medium is higher than that of LB and TSB medium containing 1% NaCl, and the maximum y...

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Abstract

The invention discloses a salt-tolerant bacteria T5S2 and applications in microbial fertilizer. A new specie salt-tolerant bacteria T5S2 can be separated from rhizosphere soil of Kalidium gracile in Sinkiang Junggar basin salt and alkali region, the salt-tolerant bacteria T5S2 can tolerate 1-15% NaCl, can grow well at 30 DEG C-37 DEG C, and can produce ACC deaminase and IAA indoleacetic acid without induction during growth, a microbial fertilizer prepared by utilizing the bacteria is applied to commercial crops of saline-alkaline land cotton, corns and the like, so that the germination rate of the saline-alkaline land cotton can achieve 77.8%, the survival rate can achieve 65.4%, the plant height, plant number and single boll plant number are obviously higher than cotton normally treated, the seedling rate of the saline-alkaline land corns can be increased to more than 80% from 64.5%, the production quantity of corn per mu is up to 600kg / mu, the total content of salt in soil of 0-30cm soil layer is reduced by 60-70%, and the microbial fertilizer has obvious salt reduction and production increase functions, thus having wide application values.

Description

technical field [0001] The invention relates to the technical field of application of microbial strains, in particular to the technical field of a salt-tolerant bacterium and its application in the preparation of microbial fertilizers. Background technique [0002] Halomonas (Halomonas) is a kind of bacteria with strong salt tolerance, most of them can grow at the concentration of 0.1%-20% NaCl, and some can also grow at the concentration of 0.1%-32.5%, usually distributed in salt lakes, Salt fields, etc., belong to moderate halophiles. They and extreme halophilic archaea (extrem eha lophiles) are both extremophiles, but they are more widely distributed than halophilic archaea, have low nutritional requirements, and are easier to adapt to the environment. The unique biological characteristics, physiological mechanisms and genetic mechanisms of Halomonas have gradually attracted the interest of researchers. [0003] At present, secondary salinization of soil caused by impro...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F11/08C12R1/01
CPCC05F11/08C12N1/205C12R2001/01
Inventor 王晓桦张謇汪玲许咏梅杨金钰冯新玉邝作玉唐鹏王蓓蓓崔艺凡王春燕唐天成
Owner 新疆山川秀丽生物有限公司
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