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A kind of Geomonas rf4 and its application

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: 2022-07-05
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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  • A kind of Geomonas rf4 and its application
  • A kind of Geomonas rf4 and its application
  • A kind of Geomonas rf4 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Sample enrichment culture

[0030] Weigh 10g of paddy soil to 90mL of sterile MFM liquid medium, N 2 : CO 2 (V / V, 80:20) deoxygenated for 0.5 h to form an anaerobic environment, and then closed with an aluminum cap and then statically enriched and cultured 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 , 1.0g NH 4 Cl, 0.1 g MgCl 2 ·6H 2 O, 0.08g CaCl 2 ·2H 2 O, 0.6g NaCl, 9.52g HEPES, 10mL mineral solution [(L -1 ): NTAT Trisodium 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 Pyr...

Embodiment 2

[0058] 16S rRNA gene identification

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

[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 PCR products and spot them on a 1% agarose gel, with 100bp Marker as the standard molecular weight, 100V voltage, electrophoresis for 30min, and observe the electrophoresis results with a gel imaging system. The PCR products of the strains with detected bands were sent to Sangon Bioengineering (Shanghai) Co., Ltd. for sequencing. ContigExpress software was used to proofread the 16S rRNA gene sequence obtained by sequencing, and the cluttered bases at the head and tail ends were removed, and the obtained effective sequence was submitted t...

Embodiment 3

[0070] Fatty acid detection

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

[0072] ②Saponification: Add 1.0 mL of saponification reagent to the test tube, tighten the lid, shake the test tube for 5-10 s with a shaker, take a boiling water bath for 5 min, take it out and continue to shake for 5-10 s, tighten the lid again, continue the water bath for 25 min, remove the test tube, and cool at room temperature.

[0073] ③Methylation: Add 2.0 mL of methylation reagent to the test tube, tighten the lid, shake with a shaker for 5-10 s, take a water bath at 80°C for 10 min, 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 quickly for 10min, open the tube cap, suck out the lower water-like phase of the test tube with a pipette and discard it.

[0075] ⑤Wa...

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PUM

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Abstract

The invention discloses a Geomonas niftrogeniifigens RF4 and its application. It belongs to the field of microbial technology. Geomonas RF4 is strictly anaerobic, gram-negative, motile, and flagella all over the body. Colonies are red and round. Contactase and oxidase reactions were negative. Strain RF4 has the ability to produce alkaline phosphatase, acid phosphatase and naphthol-AS-BI-phosphohydrolase. For the first time, Geomonas was found to have both iron-reducing and nitrogen-fixing functions. It has good application prospects in agricultural production to drive rice growth, reduce nitrogen fertilizer application, and reduce environmental pollution.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a Geomonas RF4 and its application. Background technique [0002] Rice is the largest grain crop in my country, and nitrogen is the main limiting factor for rice yield. Biological nitrogen fixation and chemical nitrogen fertilizer are the main nitrogen sources for rice growth, but the large-scale application of chemical nitrogen fertilizer 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 paddy fields under flooded state, reduce the application of chemical fertilizers, and reduce environmental pollution. [0003] Long-term chemical fertilization changes the microbial community structure in paddy soil, thereby reducing the diversity of nitrogen-fixing bacteria in paddy fields. In addition, it was also found that the key nitrogen-fixing microbial flora ...

Claims

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Application Information

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Patent Type & Authority Patents(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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