Bacillus mycoides R2 strain and application thereof in preventing and controlling plant root-knot nematodes

A technology of bacillus and plant roots, applied in the direction of application, nematicides, plant growth regulators, etc., to achieve the effects of inhibiting the incidence, alleviating the symptoms of root-knot disease, and good control effect

Inactive Publication Date: 2015-11-11
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the repellent mechanism of these two substances to nematodes

Method used

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  • Bacillus mycoides R2 strain and application thereof in preventing and controlling plant root-knot nematodes
  • Bacillus mycoides R2 strain and application thereof in preventing and controlling plant root-knot nematodes
  • Bacillus mycoides R2 strain and application thereof in preventing and controlling plant root-knot nematodes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Bacillus mycoides R2 strain fermented liquid is prepared by the following steps:

[0052] Cultivation of Bacillus mycoides R2 and selection of optimal loading volume:

[0053] Dip the Bacillus mycoides R2 strain preserved in glycerol with an inoculating loop and streak it on the LB plate to activate the strain. Pick the activated colony and streak it on the LB plate to get a single colony. The Bacillus mycoides was subjected to liquid fermentation, and the Bacillus mycoides was picked with an inoculation loop in LB liquid medium, and cultured in a shaker at 28° C. with a rotation speed of 200 rpm / min for 24 hours to obtain a Bacillus mycoides R2 seed liquid. According to the inoculum amount of 1%, the seed solution was respectively connected in the Erlenmeyer flasks equipped with 25ml, 50ml, 75ml, 100ml, 125ml LB liquid medium, and cultivated for three days at 28°C in a shaker at 200rpm / min. 8000rpm / min, centrifuge for 5min, discard the precipitate, and take the super...

Embodiment 2

[0056] Separation and purification of nematicide active substances produced by Bacillus mycoides R2 strain:

[0057] 1. Polar Gradient Extraction

[0058] 1) Take 25mL of the fermentation supernatant of Bacillus mycoides R2 strain in a centrifuge tube, add an equal volume of petroleum ether, invert and mix, centrifuge at 8000rpm / min for 5min. Draw the upper aqueous phase into another centrifuge tube, add an equal volume of petroleum ether, repeat this process 3 times, and finally collect the obtained petroleum ether phase into a Erlenmeyer flask, and rotatively evaporate the obtained dry matter with 1 mL of normal saline solution. Redissolve.

[0059] 2) Add an equal volume of chloroform to the remaining aqueous phase, mix by inversion, and centrifuge at 8000 rpm / min for 5 min. Draw the upper aqueous phase into another centrifuge tube, add an equal volume of chloroform, repeat this process 3 times, and finally collect the obtained chloroform phase into a Erlenmeyer flask, an...

Embodiment 3

[0074] Structural analysis of the nematicide active substance produced by Bacillus mycoides R2:

[0075] HPLC-MS analysis:

[0076] The liquid mass spectrometry system of Agilent Technologies 1260 and Agilent Technologies 6540 UHDAccurate-MassQ-TOFLC-MS was used for positive ion mode detection. The ion source parameters were gas temperature 350 ° C, gas flow rate 9.0 L / min, nebulizer pressure 0.2756 MPa, and the mass of the reference particle. The nuclei ratios (m / z) are 121.0509 and 922.0098, the detection range of the mass spectrometer is 100-1700m / z, and the acquisition rate is 1.5spectrac / sec. The components containing the anti-insect active substances collected by HPLC are determined. The separation column is a C18 (2.1mm×100mm) chromatographic column of Agilent Co., Ltd., USA. The sample loading was 1 μL, the mobile phase was 10%-80% acetonitrile, the time was 40 min, the elution rate was 0.5 mL / min, and the detection wavelength was 210 nm. Meplin data software was use...

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Abstract

The invention discloses a Bacillus mycoides R2 strain and application thereof in preventing and controlling plant root-knot nematodes. The Bacillus mycoides R2 strain is collected under a number of CCTCC NO: M2015251. The Bacillus mycoides R2 strain is confirmed to be Bacillus mycoides through 16S rRNA analysis and other morphological analysis and named as Bacillus mycoides R2. The Bacillus mycoides R2 strain which is screened out has higher activity in resisting nematodes, so that theories of Bacillus nematicides are enriched. N,N-diethyl-m-toluamide (DEET) and styrene which are generated by the Bacillus mycoides R2 strain has a repelling effect on nematodes, so that a theoretical foundation is laid for development of nematode repellents and the application thereof in nematode prevention and control. Interrelation and action molecule mechanism between pathogenic microorganisms and hosts are clarified, so that a theoretical foundation is laid for application of semiochemicals in the aspects of biological prevention and control of nematodes.

Description

technical field [0001] The invention relates to the fields of agricultural microbiology technology and biochemistry and molecular biology technology, in particular to a Bacillus mycoides R2 strain and its application in preventing and treating plant root-knot nematode disease. Background technique [0002] In recent years, with the adjustment of the agricultural industry structure, the planting area of ​​protected vegetables is increasing. Due to the difficulty of falling stubble and the gradual increase of the multiple cropping index, the harm of root-knot nematodes is becoming more and more serious, and the occurrence area is constantly expanding, which is one of the main limiting factors for the sustainable development of protected vegetables. Root-knot nematode (Meloidogynes pp.) is widely distributed all over the world and has a wide range of hosts. Vegetables of Solanaceae, Leguminosae, and Cucurbitaceae are the most harmed by root-knot nematodes, and the yield loss c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11A01N63/00A01P5/00C12R1/07
Inventor 赵秀云杨丽高慧娟李林
Owner HUAZHONG AGRI UNIV
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