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Bacillus aryabhattai for promoting corn root development and application thereof

A technology of Bacillus arborii and corn, applied in the field of rhizosphere growth-promoting bacteria, can solve the problems of single effect, lack of strain resources, unsuitable application, etc., to promote seedling growth and root development, improve the environment, and strengthen stress resistance sexual effect

Active Publication Date: 2020-04-10
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the first strain of Bacillus arborii was discovered and isolated in 2009, strain resources are still relatively scarce, and due to the widespread existence of Bacillus arborii, some species are not suitable for application in crops, and can only show a single effect at the same time, and There is no relevant report on the isolation of Bacillus arzii from corn, and it cannot achieve multifunctionality at the same time

Method used

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  • Bacillus aryabhattai for promoting corn root development and application thereof
  • Bacillus aryabhattai for promoting corn root development and application thereof
  • Bacillus aryabhattai for promoting corn root development and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Isolation and preliminary identification of nitrogen-fixing bacillus arborii

[0022] Soil was collected at the brown soil long-term positioning test station, and 10 g of the collected fresh soil sample was weighed, put into a shaking flask filled with 90 mL of sterile water and a small amount of small glass beads, and placed on a shaker for 30 min at 180 r / min. , so that the soil sample is evenly dispersed in the diluent to make 10 -1 Diluent. Draw 1mL 10 -1 Dilute the solution in a glass test tube filled with 9mL sterile water, shake and mix to make 10 -2 Diluent. And so on, serial dilution, until diluted to 10 -6 . This study selected 10 -3 、10 -4 、10 -5 For the three dilution gradient soil dilutions, draw 0.1mL of the dilutions and spread them evenly on the Ashby medium for separation until the surface of the solid medium dries up, and each dilution is repeated 3 times. Place the plate in a constant temperature incubator at 30°C for 5-7 days. Aft...

Embodiment 2

[0023] Molecular identification of embodiment 2 Bacillus arzii

[0024] 1. Extraction of Total DNA

[0025] Pick a single colony in Example 1 and insert it into the liquid beef extract medium, culture it at 37° C. and 180 rpm for 24 hours, take the cultured bacteria liquid and use the Ezup column bacterial genomic DNA extraction kit for DNA extraction.

[0026] 2. PCR amplification of 16S rDNA

[0027] 16S rDNA amplification primers 27F (SEQ ID NO: 1) and 1492R (SEQ ID NO: 2) were used as upstream and downstream primers, and the total DNA of LAD bacteria was used as a template for PCR amplification. The reaction system was: 1.5 μL total DNA of LAD bacteria; 12.5 μL 2×Taq MasterMix; 0.5 μL each of upstream and downstream primers (10 μmol / L); 10 μL sterile water. The reaction conditions were: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 1 min, annealing at 60°C for 1 min, extension at 72°C for 90 s, a total of 35 cycles; extension at 72°C for 10 min. The produ...

Embodiment 3

[0034] Example 3 Determination of biological properties of Bacillus aryabhattai LAD

[0035] 1. Activation of LAD strain

[0036] The preserved LAD strain was picked and cultured in beef extract-peptone liquid medium at 37°C and 180 r / min for 24 hours to make the LAD strain active.

[0037] 2. Growth curve determination

[0038] The activated bacterial solution was inserted into 50ml of beef extract peptone medium according to the inoculum amount of 2wt%, and cultured at 180r / min at 37°C. Measure the OD of the bacterial solution every 2 hours 600 Value until the OD value tends to be stable, do three parallel groups, and draw the growth curve with the blank medium as the control. As shown in the figure, the LAD strain is in the lag phase within 0-4 hours of inoculation, and enters the logarithmic phase after 4 hours. At this stage, the strain grows faster, starts to grow slowly at 12 hours, and tends to be stable at 32 hours.

[0039] 3. niLADH nitrogen fixation gene assay ...

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Abstract

The invention relates to the field of plant growth-promoting rhizobacteria (PGPR), in particular to Bacillus aryabhattai for promoting corn root system development and application of the Bacillus aryabhattai. The strain is Bacillus aryabhattai LAD, and is preserved in China General Microbiological Culture Collection Center, wherein the preservation site is No.3, Yard 1, Beichen West Road, ChaoyangDistrict, Beijing, the preservation number is CGMCC NO: 18653, and the preservation time is October 10, 2019. The invention relates to an application of the Bacillus aryabhattai in promoting corn root development. The Bacillus aryabhattai LAD can stably exist in soil, has the characteristics of nitrogen fixation, sugar production, starch decomposition, plant hormone production and the like, and promotes the growth of corn seedlings and the development of root systems. Moreover, the Bacillus aryabhattai has relatively strong stress resistance, so that the Bacillus aryabhattai can be applied tocrop production.

Description

technical field [0001] The invention relates to the field of rhizosphere growth-promoting bacteria (PGPR), in particular to a strain of bacillus arborii for promoting the development of corn roots and an application thereof. Background technique [0002] Bacillus arseni was discovered in the early 21st century. It belongs to Gram-positive bacteria and the colony of Bacillus is smooth, with a size of 5-10mm. The research direction of Bacillus arborii includes macromolecule degradation, product synthesis and promotion of plant growth. Bacillus arborii is widely distributed in nature and has strong stress resistance. It is found in animal intestines, plant rhizosphere, deep sea and plateau. Bacillus arseni can not only degrade or synthesize macromolecular compounds, including decolorization, sewage degradation, synthetic spices, synthetic biological flocculants and anti-leukemia specific drugs; it can also symbiose with plants and promote plant growth, including synthetic horm...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/22A01P21/00C09K17/14C12R1/07
CPCC12N1/20C09K17/14C12R2001/07C12N1/205
Inventor 李炳学邓超张宁张志勇丁锐
Owner SHENYANG AGRI UNIV
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