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Test method for xanthomonas axonopodis pv. citri antagonistic bacteria antibacterial capability

A technology of citrus canker and a determination method, which is applied in the field of determination of the bacteriostatic ability of antagonistic bacteria to plant pathogenic bacteria, can solve the problems of inaccurate size of the inhibition zone, difficulty in simultaneously comparing the bacteriostatic ability and the like, and achieves simple operation and experimental cost. Low, good test stability effect

Inactive Publication Date: 2012-11-07
GANNAN NORMAL UNIV
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Problems solved by technology

[0004] Among the screening methods for antagonistic microorganisms of citrus canker at present, the methods for measuring the antibacterial ability of antagonistic strains include the inhibition zone method, the Oxford cup method, and the plate confrontation method. During the determination process, the antagonistic strains continued to grow on the culture medium. Due to the different biological characteristics of different strains, the ability to grow and produce metabolites is also different, resulting in inaccurate measurement of the inhibition zone size, so it is difficult to compare the inhibition of different strains at the same time. Bacterial ability

Method used

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  • Test method for xanthomonas axonopodis pv. citri antagonistic bacteria antibacterial capability

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Embodiment

[0016] Embodiment: Use the method described in the present invention to measure the bacteriostatic ability of the antagonistic bacteria that isolates obtains to X. citri:

[0017] 1. The antagonistic bacterial strain of X. citri after separation and purification was prepared with sterile water to 1.0 × 10 6 CFU / mL fresh bacterial solution, inoculate 1 mL into a conical flask filled with 50 mL of NB liquid medium, shake and culture at 180 r / min at 28°C for 72 h, centrifuge at 12,000 r / min for 20 min, and take The supernatant was filtered and sterilized through a microporous membrane with a pore size of 0.22 μm to obtain a sterile filtrate.

[0018] The NB liquid medium consists of: 3.0 g of beef extract, 1.0 g of yeast extract, 8.0 g of peptone, 10.0 g of glucose, 3.0 g of beef extract, 1,000.0 mL of water, and a pH value of 6.8-7.0.

[0019] 2. Put a piece of sterile filter paper with a diameter of 10 mm into the prepared sterile filtrate and soak for 20 minutes.

[0020] 3...

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Abstract

The invention discloses a test method for xanthomonas axonopodis pv. citri antagonistic bacteria antibacterial capability. The method includes that an antagonistic bacteria fresh bacteria liquid is inoculated into a nutrient broth (NB) liquid medium, a supernate is extracted and filtered through a millipore filter membrane, a sterile filter paper sheet is infiltrated therein, a xanthomonas axonopodis pv. Citri preparation bacteria suspension is uniformly mixed with a nutrient agar (NA) culture medium to prepare a plate, the sterile filter paper sheet is placed at the center of the plate, and an inhibition zone diameter is measured after culturing is performed for 48 hours. According to the method, the sterile filtrate of antagonistic bacteria is utilized for measuring the inhibiting capability of the antagonistic bacteria against the xanthomonas axonopodis pv. citri, so that a method for screening antagonistic bacteria of the xanthomonas axonopodis pv. citri is provided. The test method has the advantages that the inhibiting capability of the antagonistic bacteria which are obtained by separation against the xanthomonas axonopodis pv. citri can be effectively measured, the operation is simple and convenient, the test stability is good, the measuring of the inhibition zone width is accurate, and the like. By means of the method, the antagonistic bacteria which can achieve a potential application value on citrus bacterial canker can be screened out.

Description

technical field [0001] The invention relates to a method for measuring the antibacterial ability of antagonistic bacteria to plant pathogenic bacteria, in particular to a method for measuring the antibacterial ability of citrus canker sores. Background technique [0002] Citrus canker is caused by Xanthomonas rugosa Xanthomonas axonopodis pv. citri ) caused by a quarantine bacterial disease at home and abroad, mainly infects Rutaceae Citrus, Hovenia and Kumquat plants, causing citrus leaves, fruits, branches canker spots, resulting in weak growth of trees, decline in yield and quality, resulting in Huge economic loss. [0003] The disease can invade the host through natural orifices such as stomata and lenticels or wounds caused by the outside world; local transmission through wind and rain, vector insects, agricultural operations, etc., and long-distance transmission of citrus materials such as seedlings, scions and fruits with bacteria. At present, the prevention and c...

Claims

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Application Information

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IPC IPC(8): C12Q1/20C12R1/64
Inventor 易龙
Owner GANNAN NORMAL UNIV
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