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Application of punicalagin in killing pine xylophilus

A punicalagin and pine wood nematode technology, which is applied in the directions of nematicides, applications, biocides, etc., can solve the problem that the nematicide effect of the product is not as good as that of highly toxic chemical pesticides, etc., and achieves good application prospects, low cost, and wide raw material sources. Effect

Inactive Publication Date: 2016-03-30
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although a variety of natural products with activity against pine wood nematodes have been published and reported in recent years, most of them are still less effective than highly toxic chemical pesticides.

Method used

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  • Application of punicalagin in killing pine xylophilus
  • Application of punicalagin in killing pine xylophilus
  • Application of punicalagin in killing pine xylophilus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1, the preparation of punicalagin

[0018] Dry pomegranate peel powder 100g, extract with 1 liter of ethanol at room temperature for 48 hours, repeat the extraction once, concentrate the extract under reduced pressure to obtain 7.4g of paste, dilute the paste with water 10 times and then extract with ethyl acetate, the water phase After freeze-drying, 3.7g of freeze-dried powder was obtained. The freeze-dried powder was dissolved in 10mL of 20% methanol, filtered through filter paper, put on a Sephadex LH-20 chromatography column, and eluted with 20% methanol to obtain five elution fractions of I-V, and fraction V was prepared Further separation by high performance liquid chromatography (60% methanol as eluent) yielded 51.4 mg punicalagin.

Embodiment 2

[0019] Embodiment 2, the determination of the activity of killing pine wood nematode of punicalagin

[0020] Add 50 μl of pine wood nematode suspension to 1 mL and place it in a 24-well plate, so that the concentration of pine wood nematodes in it is 1000 / mL, and add 50 μl of punicalagin solution to each well, so that the final concentration of punicalagin is 200-1000 μg / mL, mix well, take the group added with equal volume of distilled water as the negative control, incubate at 28°C in the dark for a period of time, observe under a stereomicroscope, count the number of surviving and dead nematodes, and repeat the experiment 4 times. Under the visual field, the pine xylophilus is dead if it is stiff and motionless, otherwise it is alive. Calculate the corrected mortality of B. xylophilus at 24h, 48h and 72h under different gradients, and analyze the results with SPSS17.0. The calculation formula is as follows:

[0021]

[0022] The results showed that the half-lethal conc...

Embodiment 3

[0023] Embodiment 3, the inhibitory effect of punicalagin on pine wood nematode acetylcholinesterase

[0024] Healthy pine wood nematodes were mixed with physiological saline at a ratio of 1:5 (w / v), ground quickly in an ice bath, and the extract was centrifuged at 4°C, 12,000 rpm for 30 minutes, and the supernatant was obtained as acetylcholinesterase Test solution, the activity of acetylcholinesterase is measured according to the method provided by acetylcholinesterase test box (Nanjing Jiancheng Institute of Bioengineering), its principle is that acetylcholine is hydrolyzed by acetylcholinesterase to generate choline and acetic acid, and choline and sulfhydryl Chromogen reacts to generate a yellow compound, which is analyzed by colorimetry at 412nm, and the activity of acetylcholinesterase is calculated according to the amount of choline generated. The amount of one unit of acetylcholinesterase is defined as the hydrolysis of 1.0 μmol of acetylcholine within 6 minutes under ...

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Abstract

The invention relates to the application of punicalagin in killing of pine wood nematodes; the concentration of the punicalagin is 200-1000micro-g / mL, the punicalagin is capable of effectively killing the pine wood nematodes, the punicalagin has the median lethal concentration of 332.88micro-g / mL within 72 hours for the pine wood nematodes; the punicalagin is capable of effectively inhibiting the activity of acetylcholine esterase of the pine wood nematodes so as to kill the pine wood nematodes; the punicalagin has the half inhibiting concentration of 654.92micro-g / mL. The punicalagin is obtained by being separated from pomegranate bark. The punicalagin is simple in operation method, wide in raw material source, low in cost, high in efficiency and low in toxicity, safe and environmentally friendly; the punicalagin has a good application prospect in the aspect of controlling pine wilt diseases, thus being applied to pesticide preparations for killing the pine wood nematodes.

Description

Technical field: [0001] The invention relates to the application of punicalagin in killing pine wood nematodes, and belongs to the technical field of forestry pest control. Background technique: [0002] Pine wilt disease, also known as pine wilt disease (Pinewiltdisease), is a devastating forestry disease and insect pest caused by the pine wood nematode (Bursaphelenchus xylophilus) carried and transmitted by the monochamus. It brings serious economic losses, and the disease also has a serious negative impact on the ecological environment, tourist attractions and foreign trade. Many countries in the world have listed pine wood nematode as an important quarantine object. At present, the prevention and control of pine wood nematode disease is mainly based on chemical control, but the extensive use of chemical pesticides can easily cause pine wood nematodes to develop drug resistance and cause certain pollution to the environment. An important content of research on the contro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N43/90A01P5/00
Inventor 李荣贵郭群群杜桂彩郭道森
Owner QINGDAO UNIV
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