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Mesorhizobium huizhouense and application thereof

A technology of bradyrhizobium and bacterial agents, applied in the field of microorganisms, to achieve strong stress resistance and broad application prospects

Active Publication Date: 2015-04-08
RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the bottleneck restricting the development of my country's microbial fertilizer industry is the selection of excellent strains

Method used

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  • Mesorhizobium huizhouense and application thereof
  • Mesorhizobium huizhouense and application thereof
  • Mesorhizobium huizhouense and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Isolation and identification of Bradyrhizobium RITF711 and RITF712 in Huizhou

[0029] 1. Isolation of Rhizobium

[0030] The strain was isolated from the root nodules of Acacia blackwood in Huizhou, Guangdong Province. Rinse the surface of the nodules with tap water to clean the sediment, soak them in 95% alcohol for 30 seconds, put them into 0.1% mercuric chloride solution for surface disinfection for 5 minutes, and rinse them with sterile water for 8 times. Put the root nodules after surface disinfection between two sterile glass slides, squeeze them hard to break the root nodules, use a sterile inoculation needle to dip the root nodule liquid, draw a line on the YMA plate, and culture in a constant temperature incubator at 28-30°C When colonies appear, typical colonies are picked for purification and identification. Thus, rhizobium RITF711 and RITF712 were purified.

[0031] 2. Identification of Rhizobia

[0032] Rhizobium RITF711 and RITF712 were ide...

Embodiment 2

[0056] Example 2: ACC deaminase activity of Mesorhizobium Huizhou RITF711 and RITF712

[0057] In this study, the 2,4-dinitrophenylhydrazine colorimetric method was used to detect the activity of ACC deaminase in rhizobia:

[0058] 1. Detection of α-ketobutyric acid concentration

[0059] (1) Inoculate the strain to be tested into 4 ml of LB culture medium, and culture it at 28° C. and 200 rpm for 8 to 12 hours to the mid-late logarithmic or stationary phase.

[0060] (2) Take 3ml of the bacterial suspension and divide it into two 2ml centrifuge tubes, centrifuge at 12000rpm for 2min at 4°C, and remove the supernatant. Wash the bacteria twice with 1ml M9 nitrogen-free culture medium, centrifuge at 12000rpm for 2min at 4°C, and remove the supernatant.

[0061] The 2 tubes of bacterial pellets were resuspended and combined in 3.5ml M9-ACC culture medium, and cultured in a shaker at 28°C and 200rpm for 40h.

[0062] (3) Divide 3.5ml of the bacteria liquid from step (2) into tw...

Embodiment 3

[0085] Embodiment 3, Huizhou Mesorhizobium huizhouense (Mesorhizobium huizhouense) back grafting Acacia black wood effect

[0086] 1. Bacteria culture

[0087] After being activated on the YMA slant, the Bradyrhizobium Huizhou RITF711 and RITF712 strains were transferred to the YMA liquid medium, and cultured on a shaker at 28-30°C (150 rpm) to the late logarithmic growth stage.

[0088] 2. Inoculation

[0089] The seedlings are made of Acacia acacia tissue culture seedlings, the substrate is non-sterilized yellow heart soil, a plastic pot with a diameter of 12 cm is selected, double-layer filter paper is placed on the bottom of the pot, and yellow heart soil is added to the surface of the pot 1.5 cm. Acacia tissue culture bottle seedlings with similar growth were taken, the root agar was cleaned with distilled water, and then planted in plastic pots filled with yellow core soil. One seedling was transplanted in each pot, and the seedlings were managed according to convention...

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Abstract

The invention discloses a Mesorhizobium huizhouense and application thereof. The Mesorhizobium huizhouense RITF711 is preserved in China Center For Type Culture Collection (CCTCC) on July 23, 2014, the address is: Wuhan University, Wuhan, China, and the preservation number is CCTCC NO:M2014354. The Mesorhizobium huizhouense provided by the invention is separated and screened out from Acacia melanoxylon root nodule, the strain can achieve nodulation and symbiosis with Acacia melanoxylon, Acacia aneura, Acacia implexa and other hosts, and has strong ACC deaminase activity, can effectively improve the growth and stress resistance of Acacia trees, and has broad application prospects in production of nitrogen-fixing microbial agents and bio-organic fertilizers.

Description

Technical field: [0001] The invention belongs to the field of microorganisms, and in particular relates to a Huizhou bradyrhizobium and its application. Background technique: [0002] When plants are subjected to adversity stress such as drought, flood, high and low temperature, salinity and heavy metals, and pathogenic microorganisms, they will be stressed to produce a large amount of phytohormone ethylene, and high concentrations of ethylene will inhibit the growth and development of plants. 1-aminocyclopropane-1-carboxylate deaminase (1-aminocyclopropane-1-carboxylate deaminase, referred to as ACC deaminase) is an intracellular polymerase that can inhibit the synthesis of ethylene in plants. The precursor ACC decomposes into α-ketobutyrate and ammonia. ACC deaminase is an important characteristic of bacteria to promote plant growth, which can enhance the ability of plants to resist salt stress, drought stress, heavy metal pollution, and harmful microbial infection. The ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C05F11/08C12R1/01
CPCC05F11/08C12N1/205C12R2001/01
Inventor 康丽华陆俊锟窦雅静王胜坤
Owner RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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