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Method for early diagnosis and detection of pine nematode disease

A technology for pine wood nematode disease and early diagnosis, applied in botany equipment and methods, biochemical equipment and methods, microbe measurement/inspection, etc., can solve the problems of destroying forest landscape, unsightly pine trees, labor-intensive and time-consuming, etc.

Inactive Publication Date: 2003-05-28
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method can isolate nematodes, it has obvious disadvantages: 1) a lot of bark is chopped off, and the pine tree becomes ugly, destroying the forest landscape; 2) labor-intensive and time-consuming; lab space

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Wash and peel 200g of potatoes, cut into small pieces or push them into filaments, add 1000ml of water (w:v) at a ratio of 1:5, boil for 30min. (be careful not to burn, use a water bath), filter with gauze, Filter the juice and add water to the volume before cooking. Add 200g sucrose and 17g agar to adjust the pH to about 6.

[0019] Divide the prepared culture medium into 18×180mm test tubes, each test tube contains 5ml, fasten the test tube mouth with tin platinum paper, and wrap the upper edge of the test tube wall. After 1.0kg / cm 2 Sterilize for 30 minutes, then pour it into a flat surface, cool and solidify, inoculate the Pestalotia sp., and incubate at 25°C for 3 days. When the mycelium covers about 1 / 3 of the agar surface, take it out and put it in a refrigerator at 5-10°C for later use .

[0020] For the pine tree to be detected, a circular hole with a diameter of 18mm and a depth of about 60mm is chiseled or drilled on the trunk with a chisel or a drill. Th...

Embodiment 2

[0026] The difference from Example 1 is that a liquid culture medium is used, that is, 200 g of potatoes and 20 g of sucrose are added to every 1000 ml of the solution to prepare it. The prepared liquid culture medium was divided into Erlenmeyer flasks with a capacity of 500ml, and the volume of each bottle was 100ml. Then put a cotton plug or wrap the mouth of the bottle with 8 layers of gauze. After autoclaving, after cooling, inoculate the hairy dish according to the aseptic method, shake and cultivate it on a shaker for 3 days, divide it into 18×180mm test tubes, each test tube contains 2ml, seal it with tin platinum paper, and store it vertically for later use.

[0027] Use 20 test tubes inoculated with the potato juice liquid culture medium with hairy hairs, insert them into the trunks of 20 pine trees in the forest to carry out the first batch of experiments, take them back to the laboratory after 3 days on the trees, and place them at 25°C After 4 days in the incubato...

Embodiment 3

[0030] What is different from Example 1 is that the branches of pine trees have been added in the culture medium. The method is: get the branches on the healthy pine trees without worms, process them into small short sticks with a length of 50 mm and a thickness of 3 to 5 mm, autoclave them, and then put them into in the tube.

[0031] Use this inoculation plate hairy, and add 3 test tubes of potato juice agar medium with pine branches, insert them into the trunks of 3 pine trees in the forest, bring them back to the laboratory after 3 days, and check them immediately on the same day. Nematodes were found in the branch pipes. After being placed in a 25C incubator for 4 days, nematodes were found in 2 of the tubes. The results showed that nematodes were lured from two pine trees, and the positive rate was 66.7%.

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Abstract

A process for earlierly detecting pine nematosis includes inoculating the disc fungus (Ps0202) or other microbial fungi on culture medium, drilling circular hole on the pine trunk to be detected, inserting the test tube containing said fungi in said hole, staying it in the hole for 3-5 days, and determining by microscope. Its advantages are easy operation and high sensitivity.

Description

(1) Technical field [0001] The invention relates to a method for detecting pine wood nematode disease. (2) Background technology [0002] Pine wood nematode (Busaphelenchus xylphilus) disease is a devastating disease of pine trees, which can cause large areas of pine trees to die quickly and cause huge economic losses; Losses and disease epidemics. So its early diagnosis is particularly important. [0003] Originally, the detection of pine wood nematodes followed the Baermann's funnel method. The basic device is a funnel of suitable diameter, the end of the funnel tube is connected with a section of rubber tube, and the opening and closing of the tube is controlled by a spring water stop clip. The funnel is placed on the ring of the iron frame, and it is filled with clean water. Part of the bark is split from the trunk of the pine tree, cut into small pieces, wrapped in a square gauze cloth, and soaked in water. Nematodes swim out of pine bark. The gauze sinks to the b...

Claims

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Application Information

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IPC IPC(8): A01N63/04C12Q1/02G01N33/00
Inventor 潘沧桑
Owner XIAMEN UNIV
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