Bacillus pumilus having functions of improving acid soil and decomposing potassium, and preparation and application of microbial inoculum of bacillus pumilus

A technology of Bacillus pumilus and inoculum, applied in the field of agricultural microorganisms, can solve problems affecting plant growth, root development, resistance to stress and pests and diseases, aggravate soil acidification, damage to soil structure, etc., and achieve high salt tolerance. good performance, good growth and good fermentation properties

Active Publication Date: 2019-12-31
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Potassium deficiency affects plant growth, root development, stress resistance and resistance to pests and diseases, resulting in low crop yields
At present, about 60% of the cultivated land in my country is deficient in potassium, but the soil is rich in silicate minerals. Because of its insoluble nature, potassium is difficult to release. In order to alleviate the situation of potassium deficiency in the soil, it is necessary to supplement potassium with chemical fertilizers The method commonly used in agricultural production, but long-term excessive use of chemical fertilizers leads to soil structure damage, resulting in nutrient loss and environmental pollution; moreover, after crops absorb the "potassium ions" in the fertilizer, they will leave acid radical ions such as "sulfate" and " Chloride ions, etc., these ions further aggravate the acidification of the soil

Method used

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  • Bacillus pumilus having functions of improving acid soil and decomposing potassium, and preparation and application of microbial inoculum of bacillus pumilus
  • Bacillus pumilus having functions of improving acid soil and decomposing potassium, and preparation and application of microbial inoculum of bacillus pumilus
  • Bacillus pumilus having functions of improving acid soil and decomposing potassium, and preparation and application of microbial inoculum of bacillus pumilus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1: Isolation and screening of bacterial strain

[0060] Collect wheat rhizosphere soil samples from saline-alkali land in the Yellow River Delta, take 10g soil samples under aseptic conditions, dissolve them in 90mL sterile distilled water, 220r / min, vibrate for 30min, absorb 1mL and mix them with 9mL sterile water to obtain 10 -1 diluent, then take 1mL10 -1 The diluted solution was mixed with 9mL sterile water to obtain 10 -2 Dilutions, indistinctly, to prepare 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 Wait for a series of dilutions. The chosen dilution is 10 -5 、10 -6 、10 -7 100uL of the soil dilution solution was drawn and spread on the PDA medium, and cultured at a constant temperature of 30-32°C for 1-3 days. Pick a single colony, streak it into a pure culture, and save it.

[0061] Several strains of pure cultures obtained above were subjected to a potassium-dissolving test, and the size of the potassium feldspar hydrolysis circle was measured. T...

Embodiment 2

[0067] Embodiment 2: the identification of bacterial strain

[0068] (1) Cell morphology and colony morphology

[0069] The colonies of the HG-8 strain on the PDA plate are light yellow, round and translucent, with neat edges and moist surface, with a slight bulge in the middle of the colony, and the shape of the bacteria observed under a microscope is rod-shaped.

[0070] (2) Physiological and biochemical characteristics

[0071] The physiological and biochemical characteristics of the HG-8 strain are shown in Table 1. The aerobicity of the strain is facultative anaerobic, Gram reaction, glucose oxidation fermentation test, contact enzyme reaction, starch hydrolysis test, methyl red test, phenylalanine amino acid deaminase test and citrate utilization test were positive; V-P test and nitrate reduction reaction were negative; it can grow in liquid LB liquid medium with NaCl concentration of 2% and 5%. LB liquid medium components: peptone 10g, yeast powder 5g, NaCl 10g, wate...

Embodiment 3

[0088] Example 3: HG-8 strain rep-PCR gene fingerprint.

[0089] 1. Genomic DNA extraction of HG-8 strain

[0090] TIANGEN Bacterial Genomic DNA Extraction Kit DP302 was used.

[0091] (1) Take 1 mL of HG-8 strain culture solution, centrifuge at 10,000 rpm for 1 min, and suck up the supernatant as much as possible;

[0092] (2) Add 110 μL TE buffer and 70 μL lysozyme to the cell pellet, and treat at 37°C for more than 30 minutes;

[0093] (3) Add 20 μL of Proteinase K solution to the tube and mix well;

[0094] (4) Add 200 μL of buffer solution GB and shake for 15 sec, place at 70°C for 10 min, the solution should become clear, and briefly centrifuge for 10-15 sec to remove water droplets on the inner wall of the tube cap;

[0095] (5) Add 220 μL of absolute ethanol, shake and mix well for 15 seconds, at this time flocculent precipitation may appear, briefly centrifuge for 10-15 seconds to remove water droplets on the inner wall of the tube cap;

[0096] (6) Add the soluti...

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Abstract

The invention discloses bacillus pumilus having functions of improving acid soil and decomposing potassium, and preparation and application of microbial inoculum of the bacillus pumilus. According tothe thallus shape, the colony characteristics and the physiological and biochemical properties of an HG-8 strain, through combination of the analysis result of 16S rDNA and gyrB sequences, the HG-8 strain is identified as the bacillus pumilus, the bacillus pumilus is preserved in China General Microbiological Culture Collection Center on 26 September 2019, and the preservation number is CGMCC NO.18620. The HG-8 strain can be stably colonized in the acid soil, the microbial inoculum prepared from the strain can be applied to improvement of growth of crops of wheat, corn, chilies and the like inthe acid soil, the pH value and the effective potassium content of the acid soil can be notably increased, and the yield of the crops can be increased. The strain has salt tolerance, besides, the fermentation properties of the strain are excellent, the microbial inoculum is simple in preparation technology, short in fermentation period and low in cost, and industrialized production and transportation are facilitated.

Description

technical field [0001] The invention relates to the technical field of agricultural microorganisms, in particular to the preparation and application of a strain of Bacillus pumilus with the functions of improving acidic soil and dissolving potassium and its bacterial agent. Background technique [0002] Soil acidification refers to the phenomenon that soil acidity becomes stronger under natural and artificial conditions, that is, the soil pH value decreases. one of the important performances. Soil acidification leads to a decrease in the availability of soil nutrients, enhanced activity of heavy metal ions, soil compaction, less voids, and poor air permeability, which is not conducive to the adjustment of water, air, heat, and fertilizer in the soil, resulting in soil degradation, which in turn affects the normal growth and development of crops. leading to a decline in crop yield and quality. [0003] Since the 1980s, the situation of soil acidification in my country has b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10C05G3/04C09K17/14A01N63/00A01P21/00C09K101/00C09K109/00C12R1/07
CPCA01N63/00B09C1/10C09K17/14C09K2101/00C09K2109/00C12N1/20C12N1/205C12R2001/07
Inventor 刘训理宋鑫胡玉蓉延栋梁纪超王长栋刘玥郝立平周艳艳徐睿娉
Owner SHANDONG AGRICULTURAL UNIVERSITY
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