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Salt-tolerant bacillus subtilis for producing acetoin and application of salt-tolerant bacillus subtilis

A technology of Bacillus subtilis and acetoin, which is applied in the field of microorganisms, can solve the problems of cumbersome operation, cumbersome operation process, low salt content in saline-alkali soil, etc., and achieves the effects of expanding the planting range, increasing the planting area, and improving the salt tolerance.

Active Publication Date: 2019-06-11
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Patent CN201611205485.3 discloses a method of using a special biological fertilizer for alfalfa. Its characteristic value is that the special fertilizer for alfalfa contains strains such as Bacillus subtilis. Although it improves the yield of alfalfa, it does not mention promoting the growth of alfalfa in saline-alkali land
Patent CN105296381 A discloses a strain of Bacillus subtilis CYY-25 and its application method, which is characterized in that it can promote the growth of alfalfa in saline-alkali soil, but it does not explain the salt content of saline-alkali soil, and its operation is cumbersome. During the growth process, the CYY-25 bacterial suspension needs to be poured every 7 days
Although Bacillus subtilis can promote the growth of alfalfa in saline-alkali land and non-saline-alkali land, the saline-alkali soil it is used in is relatively low in salt content and the operation process is cumbersome, so it is difficult to meet the requirements of moderate and severe saline-alkali soil

Method used

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  • Salt-tolerant bacillus subtilis for producing acetoin and application of salt-tolerant bacillus subtilis
  • Salt-tolerant bacillus subtilis for producing acetoin and application of salt-tolerant bacillus subtilis
  • Salt-tolerant bacillus subtilis for producing acetoin and application of salt-tolerant bacillus subtilis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Isolation and screening of strain NRCB002

[0026] Take 10g of rice rhizosphere soil and put it in an Erlenmeyer flask containing 90mL salt solution, shake for 1h at 28℃, 200r / min, and make 10 -1 Soil suspension, take 10 -1 10mL of the soil suspension, put it into a bottle containing 90mL salt solution, make 10 -2 Diluent, and so on. Make 10 separately -3 , 10 -4 , 10 -5 Diluent. Then use a pipette to remove 10 -3 , 10 -4 , 10 -5 100μL of the diluted solution was spread on the nitrogen-fixing solid medium, and each gradient was repeated in three times. Cultivate for 2 to 4 days in a constant temperature biochemical incubator at 28°C. Observe its growth every day. If single bacteria are lagging behind, use an inoculation loop to pick larger colonies on the BMS medium for purification and purify for 5 to 5 days. After 6 generations, store it at -80°C with 30% glycerol for later use.

Embodiment 2

[0027] Example 2 Identification of strain NRCB002

[0028] See the appearance of the strain figure 1 .

[0029] 16S rDNA identification through 16S rDNA sequence analysis, after BLAST alignment in NCBI, it has a higher homology with Bacillus subtilis, and the similarity is 100%. Based on its appearance and physiological and biochemical characteristics, the strain NRCB002 is preliminarily identified as Bacillus subtilis.

[0030] Physiological and biochemical properties are shown in Table 1: Starch hydrolysis, V-P reaction, gelatin liquefaction, glucose acid production, lysozyme, and methyl red tests are all positive.

[0031] Table 1 Physiological and biochemical properties of strain NRCB002

[0032] Strain number

Embodiment 3

[0033] Example 3 Analysis of salt tolerance of Bacillus subtilis NRCB002

[0034] The isolated and purified salt-tolerant strain Bacillus subtilis NRCB002 suspension was transferred to the same amount of liquid medium with different salt concentration, 25~30℃, 150~200r / min shaker culture, and the growth was observed. Measure the bacterial concentration (OD600) within 1d to determine the salt tolerance of the bacteria. See the salt tolerance of the strain figure 2 . It can be seen from the figure that as the concentration of NaCl increases, the strain NRCB002OD600 gradually increases, which can infer that the strain NRCB002 is a halophilic bacteria.

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Abstract

The invention discloses salt-tolerant bacillus subtilis capable of producing acetoin. The strain number is NRCB002, the strain is preserved in the China General Microbiological Culture Collection Center, the preservation number is CGMCC NO.17213, and the preservation date is January 18, 2019. The invention also discloses an application of the bacillus subtilis. Pure culture tests, laboratory platetests and greenhouse pot experiments prove that the bacillus subtilis NRCB002 has salt tolerance, can produce acetoin, can promote growth of alfalfa, and has a good application prospect.

Description

Technical field [0001] The invention belongs to the technical field of microorganisms and relates to Bacillus subtilis and its application in promoting crop growth. Background technique [0002] At present, in agricultural production, due to the long-term unreasonable use of agricultural inputs such as chemical fertilizers, not only the acidification, compaction, and secondary salinization of farmland soil have become more and more serious, but also the structure of the soil rhizosphere microbial community is destroyed, and many beneficial microorganisms are removed from the soil. The dominant flora becomes the secondary flora. Alfalfa (Medicago sativa L.) is the most widely distributed perennial leguminous forage in the world, and is known as the "king of forage". The planting area of ​​alfalfa in my country is about 3.775 million ha 2 However, the output of alfalfa is low, and alfalfa is imported from abroad every year to meet the needs of domestic animal husbandry. Alfalfa h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/26A01G22/40C12R1/125
Inventor 高南朱志玉应汉杰陈晓春陈勇牛欢青柳东
Owner NANJING UNIV OF TECH
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