Acinetobacter calcoaceticus with free living nitrogen fixation and phosphorus and potassium dissolving capability and application of acinetobacter calcoaceticus

A technology of calcium acetate, ability, applied in the field of agricultural microorganisms

Active Publication Date: 2014-05-14
西安金博瑞生态科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the nitrogen-fixing bacteria strains currently used in production in my country have low nitrogen-fixing capacity and unstable application effects, and most of the wild nitrogen-fixing bacteria isolated from nature have slow growth, weak viability, short validity period, nitrogen fixation and growth-promoting ability The shortcomings such as low and harsh requirements on the soil environment cannot meet the needs of growth-promoting microbial fertilizers

Method used

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  • Acinetobacter calcoaceticus with free living nitrogen fixation and phosphorus and potassium dissolving capability and application of acinetobacter calcoaceticus
  • Acinetobacter calcoaceticus with free living nitrogen fixation and phosphorus and potassium dissolving capability and application of acinetobacter calcoaceticus
  • Acinetobacter calcoaceticus with free living nitrogen fixation and phosphorus and potassium dissolving capability and application of acinetobacter calcoaceticus

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The nitrogen-fixing bacterial strain of the present invention is obtained by the method of ultraviolet-plasma compound mutagenesis, and the specific steps are as follows:

[0044] 1) Use the Agrobacterium SGD07 screened from the plant rhizosphere in the laboratory as the starting strain;

[0045] 2) Mutation breeding

[0046] (1) Prepare the single cell suspension of the starting strain SGD07

[0047] The starting strain SGD07 was inoculated in liquid medium A, cultured at 28°C and 120rpm for 16 hours, centrifuged, washed with sterile saline, placed in a conical flask with glass beads, and oscillated to disperse into single-cell bacteria suspension;

[0048] Described medium A consists of: yeast powder 5g, NaCL 5g, peptone 5g, FeSO 4 ·7H 2 O 0.01g, Na 2 MoO 4 2H 2 O 0.005 g, distilled water 1000ml, pH 7.2.

[0049] (2) Ultraviolet mutagenesis

[0050] Adjust the concentration of the bacterial suspension obtained in step (1) to 10 7 1 / ml, take 0.1ml and smear ...

Embodiment 2

[0072] Example 2 Determination of the ability of SGD07-3-16 to produce indole acetic acid (IAA)

[0073] Add a concentration of 200 μg·ml to a 250 ml Erlenmeyer flask containing 100 ml of liquid medium A -1 Inoculate SGD07-3-16 in 50ml of liquid medium A, culture at 28°C and 150rpm for 20 hours, take 100 μl (dilute the bacterial solution with medium A to make it OD 600 0.7) The bacterial solution was inoculated in the above-mentioned liquid medium A containing L-tryptophan, cultured at 28°C, 150 rpm for 72 hours, centrifuged at 4000 g for 10 min, and the concentration of IAA in the supernatant was measured by spectrophotometry. The experimental results showed that: SGD07-3-16 produced IAA concentration of 36.72μg / ml.

Embodiment 3

[0074] Example 3 Determination of ACC Deaminase Activity Produced by SGD07-3-16

[0075] After inoculating SGD07-3-16 in 50ml liquid medium A for 20 hours of activation, take 100 μ l (dilute the bacterial solution with medium A to make it OD 600 0.7) Bacterial solution was inoculated in 100ml of liquid medium A, cultured at 28°C, 150rpm for 24 hours, centrifuged to collect the bacterial cells, at 4°C with DF culture medium (without (NH 4 ) 2 SO 4 ) washed twice, the cells were suspended in ADF culture medium, cultured at 28°C and 150rpm for 24 hours to induce the production of ACC deaminase, the cells were collected by centrifugation at 4°C, and 0.1 mol / L Tris-HCl buffer (pH 7.6 ) wash twice, resuspend in 600 μl 0.1mol / L Tris-HCl buffer (pH 8.5), add 30 μl toluene, and shake rapidly for 30s to break the cells, transfer 200 μl cell extract containing toluene After adding 20 μl of 0.5mol / L ACC and mixing, the ACC deaminase activity was measured. The results showed that the...

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Abstract

The invention relates to acinetobacter calcoaceticus X.L.Zhu.SGD07-3-16 with free living nitrogen fixation capability. The acinetobacter calcoaceticus is preserved in China general microbiological culture collection center, and the preservation number of the acinetobacter calcoaceticus is CGMCCNo.5455. The acinetobacter calcoaceticus is high in nitrogenase activity and phosphorus and potassium dissolving capability, and has ACC (1-aminocyclopropane-1-carboxylic acid) deaminase activity, a certain amount of indoleacetic acid can be generated, and the generation of siderophore is avoided; when an acinetobacter calcoaceticus-containing liquid or solid microbial agent is used for inoculating crops such as corns, oilseed rapes and leaf mustard, the growth of the crops can be remarkably promoted, and the yield of the crops can be increased.

Description

technical field [0001] The invention relates to a strain of Acinetobacter calcoacetate with the ability of autogenous nitrogen fixation and phosphorus and potassium dissolution. The strain is obtained by ultraviolet-plasma compound mutagenesis, can promote the growth of crops, and belongs to the technical field of agricultural microorganisms. technical background [0002] Nitrogen is one of the macroelements necessary for crop growth and development, and contributes 40%-50% to the final yield of crops. Crops take away a large amount of nitrogen from the soil every year. In order to supplement the loss of soil nitrogen, industrial chemical nitrogen fertilizers are still an important source of human nitrogen supply to the soil. Although chemical fertilizers have played a huge role in promoting crop growth and increasing food production, long-term excessive use of chemical fertilizers will cause a series of problems, such as acid deposition, soil compaction, water eutrophicatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N15/01C05G3/00C12R1/01
Inventor 朱晓丽马俊杰宋进喜梁丽华李贺
Owner 西安金博瑞生态科技有限公司
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