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Geomonas RF4 and application thereof

A technology of bacterium and RF4, applied in the field of microbiology, can solve the problem of few research reports on Geomonas resources

Active Publication Date: 2021-09-03
福建省农业科学院农业生物资源研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, Geomonas only contains 9 effective species, and there are few reports on the resources of Geomonas

Method used

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  • Geomonas RF4 and application thereof
  • Geomonas RF4 and application thereof
  • Geomonas RF4 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Sample enrichment culture

[0030] Weigh 10g paddy soil to 90mL sterile MFM liquid medium, N 2 :CO 2 (V / V, 80:20) Deoxygenation for 0.5h to form an anaerobic environment, after sealing the aluminum cap, enrich and culture at 30°C for 2 weeks.

[0031] MFM medium (L -1 ): 2.0g KHCO 3 , 0.02g MgSO 4 ·7H 2 O, 0.3g KH 2 PO 4 , 1.0 g NH 4 Cl, 0.1gMgCl 2 ·6H 2 O, 0.08g CaCl 2 2H 2 O, 0.6g NaCl, 9.52g HEPES, 10mL mineral solution [(L -1 ): NTATrisodium Salt (Free acid) 1.50g, MgSO 4 3.00g, MnSO 4 ·H 2 O 0.50g, NaCl 1.00g, FeSO 4 ·7H 2 O 0.10g, CaCl 2 2H 2 O 0.10g, CoCl 2 ·6H 2 O 0.10g, ZnCl 2 0.13 g, CuSO 4 ·5H 2 O 0.01g, AlK(SO 4 ) 2 12H 2 O 0.01g, H 3 BO 3 0.01 g, NaMoO 4 2H 2 O 0.09g], 10mL vitamin mixture [(L -1 ): biotin 0.002g, pantothenic acid 0.005g, vitamin B12 0.0001g, p-aminobenzoic acid 0.005g, lipoic acid (α-) 0.005g, niacin 0.005g, thiamine 0.005g, riboflavin 0.005g, hydrochloric acid Pyridoxine 0.01g, folic acid 0.002g], pH 6.8. ...

Embodiment 2

[0058] 16S rRNA gene identification

[0059] The isolated strains were amplified by bacterial 16S rRNA gene universal primers 27F (5′-GAG TTTGAT CCT GGC TCA G-3′) and 1492R (5′-ACG GCT ACCTTG TTA CGA CTT-3′).

[0060] PCR reaction program: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, annealing at 55°C for 60 s, extension at 72°C for 90 s, a total of 30 cycles; final extension at 72°C for 10 min. Take 5 μL of the PCR product, spot on 1% agarose gel, use 100 bp Marker as the standard molecular weight, 100 V voltage, electrophoresis for 30 min, and observe the electrophoresis result with a gel imaging system. The PCR products of strains with bands detected were sent to Sangon Bioengineering (Shanghai) Co., Ltd. for sequencing. ContigExpress software was used to proofread the 16S rRNA gene sequence obtained by sequencing, remove the cluttered bases at both ends, and submit the obtained effective sequence to EZBioCloud (https: / / www.ezbiocloud.net / ) and NCBI (...

Embodiment 3

[0070] Fatty acid detection

[0071]① Bacteria acquisition: Pick about 40 mg of bacteria in the logarithmic phase (cultured for 3 days) and put them into a test tube with a screw cap (specification 13mm×100mm).

[0072] ②Saponification: Add 1.0mL of saponification reagent to the test tube, tighten the cap, shake the test tube with an oscillator for 5-10 seconds, bathe in boiling water for 5 minutes, take it out and continue shaking for 5-10 seconds, tighten the cap again, continue the water bath for 25 minutes, remove the test tube, and cool at room temperature.

[0073] ③Methylation: Add 2.0mL of methylation reagent to the test tube, tighten the cap, oscillate with an oscillator for 5-10 seconds, place in an 80°C water bath for 10 minutes, remove the test tube and quickly rinse with tap water to cool to room temperature.

[0074] ④Extraction: Add 1.25mL of extraction reagent, tighten the cap, shake rapidly for 10 minutes, open the cap, suck out the lower layer of the test tub...

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Abstract

The invention discloses a Geomonas niftrogenia RF4 and application of the Geomonas niftrogenia RF4, and belongs to the technical field of microorganisms. The Geomonas niftrogenia RF4 is strictly anaerobic, gram-negative, moving, and flagellar in the whole body. The bacterial colony is red and round. The reaction of catalase and oxidase is negative. The strain RF4 has the capability of producing alkaline phosphatase, acid phosphatase and naphthol-AS-BI-phosphohydrolase. It is found for the first time that geomonas has the functions of iron reduction and nitrogen-fixing bacteria at the same time. In agricultural production, rice growth promotion is driven, nitrogen fertilizer application is reduced, environmental pollution is reduced, and good application prospects are achieved.

Description

technical field [0001] The invention relates to the technical field of microbes, and more specifically relates to Geomonas RF4 and its application. Background technique [0002] Rice is the largest food crop in my country, and nitrogen is the main limiting factor of rice yield. Biological nitrogen fixation and chemical nitrogen fertilizers are the main nitrogen sources for rice growth, but excessive application of chemical nitrogen fertilizers will bring a certain degree of environmental pollution. Biological nitrogen fixation is an important process in paddy fields, which can maintain the balance of nitrogen in the flooded state of paddy fields, reduce the application of chemical fertilizers, and reduce environmental pollution. [0003] Long-term chemical fertilization caused changes in the structure of rice soil microbial communities, thereby reducing the diversity of nitrogen-fixing bacteria in rice fields. In addition, it was also found that the key nitrogen-fixing mic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/16A01N63/20A01P21/00C05F11/08C12R1/01
CPCC12N1/20C12N9/16A01N63/20C05F11/08C12Y301/03001C12Y301/03002Y02P60/21
Inventor 刘国红周顺桂杨尚唐荣
Owner 福建省农业科学院农业生物资源研究所
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