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Chemical dyeing method for differentiating diffuse type pine wood nematode

A pine wood nematode and chemical dyeing technology, which is applied in the field of chemical dyeing for identifying diffusive pine wood nematodes, can solve the difficulty in counting the ratio of reproductive and diffusing nematodes, the difficulty in distinguishing reproductive and diffusing nematodes, and the complicated operation of biochemical identification methods. and other problems, to achieve the effect of simple and reliable operation, easy purchase and stable structure

Inactive Publication Date: 2007-09-26
INST OF ZOOLOGY CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, during the investigation and cultivation observation of port quarantine and forest occurrence sites of pine wood nematodes, the identification of reproductive and spreading nematodes is based on the observation of the characteristics of the nematodes under the microscope: investigators are required to have basic knowledge of the nematode structure, Moreover, it is difficult to distinguish reproductive and diffuse nematodes, especially when calculating the ratio of reproductive and diffuse nematodes.
In addition, there are currently no reliable indicators for the preliminary identification of pine xylophilus. Some reliable biochemical identification methods are complicated to operate and require related equipment and conditions, which are not easy to carry out at the grassroots level and in the field.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-3

[0022] Stimulate 1.5 μg dissolved in 20 μL of dimethyl sulfoxide (DMSO) according to the weight ratio listed in Table 1, then add 100 μL of the aqueous solution containing B. xylophilus and add 10 μL of 0.2% brillant-blue P-250 (Coomassie Brilliant Blue) was placed at room temperature for 24 hours (each treatment was repeated 10 times); and the observed staining effects were listed in Table 1.

[0023] Diffuse pine wood nematodes were treated and the staining effect was observed. Listed in Table 1. Each treatment was repeated 10 times.

[0024] α-pinene

Embodiment 4-6

[0026] Stimulate 0.5 μg of triethylene glycol diacetate dissolved in 20 μL of triethylene glycol diacetate according to the weight ratio listed in Table 2, then add 100 μL in the aqueous solution containing pine xylophilus and add 10 μL of 0.2% brillant-blueP-250 (library Marcy Brilliant Blue) was placed at room temperature for 24 hours (each treatment was repeated 10 times); and the observed staining effects were listed in Table 2.

[0027] Diffuse pine wood nematodes were treated and the staining effect was observed. Listed in Table 2. Each treatment was repeated 10 times.

[0028] α-pinene

Embodiment 7-9

[0030] Stimulant 10.5 μg dissolved in 20 μL of dichloromethane according to the weight ratio listed in Table 3, then added to 100 μL of the aqueous solution containing pine wood nematode and added 10 μL of 0.2% brillant-blue P-250 (Coomassie brilliant blue) Place at room temperature for 24 hours (10 repetitions per treatment); and the observed staining effects are listed in Table 3.

[0031] Diffuse pine wood nematodes were treated and the staining effect was observed. Listed in Table 3. Each treatment was repeated 10 times.

[0032] α-pinene

[0033] It can be seen from the table that using the chemical staining method provided by the present invention for identifying diffuse pine wood nematodes has a good identification effect, and provides a new method for the quarantine and investigation of pine wood nematodes in my country.

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Abstract

This invention relates to one chemical dyeing method to identify the diffusion of breeding pine wire worm, which comprises the following steps: a, solving the stimulant into dimethyl sulfoxide liquid with two weight proportion as 2-50:100 and element A, B and C weight proportion as 54-90:9-43:1-5 with A for alpha- firpene, B for beta- firpene and C for longifolene; b, dripping the stimulant into the water liquid with the worm and adding 10 mu L coomassie brilliant blue with intensity of 0.2% and putting in room temperature for 24 hours and observing under microscopes for 24 hours.

Description

technical field [0001] The invention relates to a chemical dyeing method for identifying diffuse pine wood nematode, in particular to a chemical dyeing method for monitoring and identifying diffuse pine wood nematode disease. Background technique [0002] Pine wood nematode disease, commonly known as "cancer" of pine trees, once a pine tree is infected with this disease, it will die in as long as half a year and as short as two months. Since the disease was first discovered near Sun Yat-sen Mausoleum in Nanjing, my country in 1982, it has spread to more than 10 provinces and cities including Jiangsu, Anhui, Guangdong, Guangxi, Zhejiang, Fujian, Shanghai, Chongqing, Hubei, Shandong, and 86 counties and cities. The area reaches 1.26 million mu, posing a serious threat to the 500 million mu of pine forest in the south. And the scope is still expanding year by year. Statistics from the State Forestry Administration show that more than 6 million pine trees are killed every year,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/29G01N31/22
Inventor 孙江华高飞韦卫赵莉蔺
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
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