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, microbial measurement/inspection, etc., can solve problems such as labor-consuming and time-consuming, occupying laboratory space, and destroying forest landscapes
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Embodiment 1
[0017] 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.
[0018] 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, and inoculate the Pestalotia sp. in the plate, and incubate at 25°C for 3 days. When the mycelium covers about 1 / 3 of the agar plane, take it out and place it in a refrigerator at 5-10°C. spare.
[0019] 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 d...
Embodiment 2
[0025] 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.
[0026] 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
[0029] 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.
[0030] 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 25°C incubator for 4 days, nematodes were found in two 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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