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A kind of preparation method and application of a solitary extract with activity of killing pine xylophilus

The technology of pine wood nematode and extract is applied in the directions of botanical equipment and method, nematicide, application, etc., and achieves the effects of simple operation method, convenient source of raw materials and low cost

Inactive Publication Date: 2019-11-12
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At home and abroad, there are no reports about the killing activity of lovage extracts on pine wood nematodes

Method used

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  • A kind of preparation method and application of a solitary extract with activity of killing pine xylophilus
  • A kind of preparation method and application of a solitary extract with activity of killing pine xylophilus
  • A kind of preparation method and application of a solitary extract with activity of killing pine xylophilus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1, the preparation of the solitary extract of killing pine xylophilus

[0026] Grind 50g of dried cloves in a plant pulverizer, add 1L of absolute ethanol to leaching with ultrasonic vibration at room temperature for 48 hours in the dark, vacuum filter to remove slag, obtain ethanol extract, repeat the extraction once, combine the extracts, and use Rotary evaporator concentrated under reduced pressure to obtain 6.3 g of brown ethanol crude extract; the ethanol crude extract was separated and extracted 3 times with ethyl acetate (1 L) and distilled water (1 L), and the ethyl acetate phase was combined and then concentrated under reduced pressure to about 2.5 g, to obtain the ethyl acetate extract of lovage.

Embodiment 2

[0027] Embodiment 2, the determination of the activity of killing pine wood nematodes of the lovage extract

[0028] Dissolve the ethyl acetate extract of lovage in 5% DMSO solution (containing 0.5% TritonX-100), prepare a solution with a concentration of 1 mg / mL, add 50 μL to the corresponding wells of the 24-well plate, and then add 50 μL The nematode suspension (about 2 / μL) was mixed evenly, and 5% DMSO solution (containing 0.5% TritonX-100) was used as the control, and each treatment was repeated 4 times. Place them in a 26°C incubator and culture them in the dark, take them out after 24 hours, 48 ​​hours, and 72 hours respectively, and observe the survival of nematodes under a stereomicroscope (nematodes are stiff and unresponsive to touch with an anatomical needle to determine the dead state) according to the following formula Calculate the corrected mortality rate of nematodes for each treatment:

[0029]

[0030] The results showed that the corrected mortality rate...

Embodiment 3

[0031] Example 3, Separation of Active Components Killing Pine Xylophilus in the Lovage Extract

[0032] 2.5 g of the ethyl acetate extract of the clodosan was subjected to silica gel column chromatography (200-300 mesh, 200 g), and gradient elution was performed using a petroleum ether-ethyl acetate solvent system (petroleum ether:ethyl acetate volume ratio=15:1, 10:1,9:1,8:1,6:1,5:1,4:1,2:1,1:1,1:3,1:4; 1:6,1:8,1: 10, 100% ethyl acetate), the elution fraction was subjected to silica gel thin layer chromatography (GF 254 ) detection, merge the same, get 10 components: Fr.1-Fr.10, through necrocidal activity test, it is determined that F5 and F6 are highly active components (1mg / mL, 72-hour corrected mortality rate > 50%), for The two components were further separated and purified; the F5 component was subjected to silica gel column chromatography and eluted with petroleum ether-ethyl acetate (8:1, v / v) to obtain two components, which were separated in petroleum ether-ethyl a...

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Abstract

The invention relates to a method for preparing a radix angelicae pubescentis extract with a pine wood nematode killing activity and an application thereof. The method comprises the following steps: taking dry radix angelicae pubescentis powder; extracting with absolute ethyl alcohol under room temperature; performing reduced pressure distillation and concentration on the extracting solution, thereby obtaining a brown rough extract; extracting a rough ethyl alcohol extract with ethyl acetate-water extracting solution for three times, merging with ethyl acetate phase; reducing the pressure and distilling, thereby obtaining the radix angelicae pubescentis extract with higher pine wood nematode killing activity; utilizing the separating methods, such as column chromatography and recrystallization, to separate four coumarins compounds with obvious pine wood nematode killing activity, namely cnidium lactone, columbianadin, bergapten and xanthotoxin, from the radix angelicae pubescentis extract. The radix angelicae pubescentis extract in the concentration of 1mg / mL has a 95.27% correcting death rate for the pine wood nematode after 72 hours. The invention has the advantages of simple operation method, convenience in raw material resource, low cost, high efficiency, low toxicity, safety and environmental protection. The radix angelicae pubescentis has a developing potential at the aspect of controlling the pine wood nematode and can be applied to a natural preparation for killing the pine wood nematode.

Description

Technical field: [0001] The invention relates to a preparation method and application of a solitary extract with the activity of killing pine wood nematodes, and belongs to the technical field of forestry pest control. Background technique: [0002] Pine wood nematode disease, also known as pine wilt disease (PWD), is a devastating pine tree disease. Once a pine tree is infected, it spreads rapidly, involving dozens of species of Pinus species. There are epidemics in many countries around the world. . Pine wood nematode disease was first discovered in Nanjing Zhongshan Cemetery Scenic Area in 1982 in mainland my country, and has now expanded to Jiangsu, Anhui, Zhejiang, Fujian, Jiangxi, Shandong, Henan, Hubei, Hunan, Guangdong, Guangxi, Chongqing, Sichuan, Guizhou, In 16 areas including Yunnan and Shaanxi, it spread rapidly, the area of ​​occurrence continued to expand, and the degree of damage continued to increase, posing a serious threat to my country's pine resources, na...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N65/10A01N43/16A01N43/90A01P5/00
CPCA01N43/16A01N43/90A01N65/00A01N65/10
Inventor 郭群群李荣贵杜桂彩梁晨燕李咏幸张明学
Owner QINGDAO UNIV
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