Method for inhibiting azotobacter in intestinal canal of termite by utilizing metabolic products of tree endophyte

A metabolite inhibition, endophyte technology, applied in microorganism-based methods, biochemical equipment and methods, and microbial determination/inspection, etc., can solve problems such as large toxic side effects, environmental pollution, human harm, etc. Long, small environmental impact, good killing effect

Inactive Publication Date: 2012-03-28
HEILONGJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The present invention aims to solve the problem that the existing method of using chemical agents to kill termites has large toxic and side effects, causing serious environmental pollution and human harm, and provides a method for inhibiting intestinal nitrogen-fixing bacteria of termites by using metabolites of endophytic bacteria in trees

Method used

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specific Embodiment approach 1

[0011] Specific embodiment one: the present embodiment utilizes tree endophyte metabolites to suppress the method for termite intestinal nitrogen-fixing bacteria, proceed according to the following steps:

[0012] 1. Pick a ring of tree endophytes, inoculate it into beef extract peptone medium, and cultivate it at 37°C and 180r / min for 2-3 days to obtain a culture solution of tree endophytes; Centrifuge the culture solution at 5000r / min for 10-15min, take the supernatant, and concentrate it to one-twentieth of its volume under the conditions of aseptic, 35°C constant temperature water bath and 150r / min to obtain the endophyte fermentation broth 2. Prepare plain agar plates: add 20 g of agar to 1000 mL of water, sterilize to obtain plain agar, then pour 10 to 15 mL of plain agar into each plate to obtain plain agar plates, and stand the Oxford cup on the plain agar plates; 3. Preparation of bacterial suspension: pick a ring of termite intestinal nitrogen-fixing bacteria, inocul...

specific Embodiment approach 2

[0018] Specific embodiment two: the difference between this embodiment and specific embodiment one is: the nitrogen-free slant culture medium described in step three is made of 15g sucrose, 0.8g KH 2 PO 4 , 0.2g MgSO 4 , 0.2g NaCl, 1g CaCO 3 , 20g agar, 1mL mass concentration of 10% Na 2 MO 4 solution, 1 mL of H 3 BO 3 solution, 1 mL of MnSO 4 solution, 1 mL of Fe 2 SO 4 solution and 1000mL H 2 O composition, adjust the pH to 6.5-7.0, and sterilize at 121°C for 30 minutes. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0019] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is: the nitrogen-free liquid culture medium described in step four is made of 15g sucrose, 0.8g KH 2 PO 4 , 0.2g MgSO 4 , 0.2g NaCl, 1g CaCO 3 , 1mL of Na with a mass concentration of 10% 2 MO 4 solution, 1 mL of H 3 BO 3 solution, 1 mL of MnSO 4 solution, 1 mL of Fe 2 SO 4 solution and 1000mL H 2 O composition, adjust the pH to 6.5-7.0, and sterilize at 121°C for 30 minutes. Others are the same as in the first or second embodiment.

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Abstract

The invention relates to a method for inhibiting azotobacter in intestinal canal of termite by utilizing metabolic products of tree endophyte, aiming at solving the problems of serious environmental pollution and serious harm to the human body as the toxic and side effects of the existing method for killing termite by utilizing chemical agent is great. The method comprises the following steps: preparing tree endophyte fermentation broth; preparing a plain agar panel; preparing bacteria suspension solution; preparing a semisolid medium; taking the nitrogen-free semisolid media, adding the bacteria suspension solution after the nitrogen-free semisolid medium is cooled to 50 DEG C, mixing to be uniform, pouring the mixer onto the plain agar panel inserted with an Oxford cup, pulling out the Oxford cup after the culture medium is solidified, adopting a liquid relief gun to add the tree endophyte fermentation broth into a small hole, putting the mixture in a fridge to stay overnight, then culturing for 16 hours in an incubator of 30 DEG C, and observing and measuring the diameter of an inhibition zone with a vernier caliper, thus completing the process. In the method, the metabolic products of tree endophyte have better inhibition action on azotobacter in intestinal canal of termite.

Description

technical field [0001] The invention relates to a method for inhibiting termite intestinal nitrogen-fixing bacteria by utilizing metabolites of tree endophytic bacteria. Background technique [0002] Termite belongs to Arthropoda phylum Insecta subclass Isoptera insects, and is a relatively ancient social pest. Termite is a typical oligotrophic organism, and the nitrogen fixation of intestinal nitrogen-fixing microorganisms is the main source of nitrogen in termites. source. The economic losses caused by termite damage to buildings and crops amount to more than 100 billion U.S. dollars every year. Existing medicaments for killing termites are all chemical medicaments, so they all have very large toxic and side effects, causing serious environmental pollution and human harm. The use of biotechnology to develop termite biological control agents with high efficiency, non-toxicity, long residual effect and little impact on the environment has become the main direction of termi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/18C12R1/065
Inventor 赵凯周东坡张小燕
Owner HEILONGJIANG UNIV
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