Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A method for high-throughput screening of pine wood nematode inhibitors

A technology for pine xylophilus and inhibitors, applied in the field of high-throughput screening of pine xylophilus inhibitors, can solve the problem of high cost

Active Publication Date: 2021-08-13
ZHEJIANG FORESTRY ACAD
View PDF8 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this kind of trunk injection is still expensive, and it can only be applied to ancient and famous trees.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A method for high-throughput screening of pine wood nematode inhibitors
  • A method for high-throughput screening of pine wood nematode inhibitors
  • A method for high-throughput screening of pine wood nematode inhibitors

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0029] Example 1. Nematode Isolation and Culture

[0030] (1) Prepare PDA medium: Take 200 grams of peeled potatoes, cut them into small pieces, add 1000 ml of water and boil for 30 minutes, filter the potato pieces with 4 layers of gauze, make up the filtrate to 1000 ml, add 20 grams of glucose, 15 grams of agar grams, after melting, subpackage, 15 pounds of sterilization (ie 121 degrees) for 15 minutes. Prepare a PDA plate on a 9cm sterile petri dish, inoculate Botrytis cinerea with a 0.5cm hole puncher, culture it to cover the entire plate at 25°C, and place it upside down at 4°C for later use;

[0031] (2) Cut the large piece of pine wood into small pieces of about 0.5*3cm, wrap it with gauze, put it in a sealed plastic, add sterile water, soak it in a water bath at 20-25°C overnight, and filter the soaking solution with 2-4 layers of gauze , the filtrate was centrifuged at 1000g for 1-5min, the supernatant was removed, and 1-5ml of worm liquid was left;

[0032] (3) Add...

example 2

[0033] Example 2. Nematode Screening

[0034] (1) Use a 1mL pipette to draw 1mL of sterile water and add it to a 9cm plate with nematodes, so many times, wash the nematode plate, and then collect the washed worm liquid into a sterile beaker;

[0035] (2) After mixing the collected worm liquid, filter it through a 10-20 μm nylon filter membrane. The filtrate is confirmed to be larvae liquid by 10*40 times microscope inspection, and those remaining on the filter membrane are adults. Remove the filter membrane from the micropore Take out from the filter, rinse in sterile water, so that the required adult liquid is obtained, and the imago liquid is used here for chemical testing;

[0036] (3) Centrifuge the worm solution obtained above at 1000g for 5 minutes at low speed, remove part of the supernatant, adjust the solution to 0.5 agarose concentration with 1 / 1000 agarose solution, so that the nematodes are evenly dispersed, and observe with a 40 times microscope, Adjust the numbe...

example 3

[0038] Example 3. Preparation and data processing of several biopesticide test panels

[0039] (1) Prepare the reagents in a 96-well plate according to the formula in Table 1 below:

[0040] Table 1. Test Plate Reagent Recipe

[0041] Group Name 1, 2, 3, 4 5, 6, 7, 8 9, 10, 11, 12

[0042] A I 25μL+25μL water J 25μL+25μL water K 25μL+25μL water

[0043] B L 25μL + 25μL water M 25μL + 25μL water N 25μL + 25μL water

[0044] C L 25μL + 25μL water M 25μL + 25μL water N 25μL + 25μL water

[0045] D L 25μL + 25μL water M 25μL + 25μL water N 25μL + 25μL water

[0046] E L 25μL + 25μL water M 25μL + 25μL water N 25μL + 25μL water

[0047] F L 25μL + 25μL water M 25μL + 25μL water N 25μL + 25μL water

[0048] G L 25μL + 25μL water M 25μL + 25μL water N 25μL + 25μL water

[0049] H 50 μL water

[0050] in FIG. 1:

[0051] A. positive control; B. test sample 1; C. test sample 2; D. test sample 3;

[0052] E. Test sample 4; F. Test sample 5; G. Test sample 6; H. Negative co...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the field of biological pest control, in particular to a method for high-throughput screening of pine wood nematode inhibitors. Pine is the most important tree species for afforestation in my country, but pine wood nematode has been the number one forest disease and insect pest for more than 30 years. The damage is very serious, and the control drugs are limited. prevention and control. The invention aims to provide technical support for screening more pine wood nematode control agents. The above-mentioned method for high-throughput screening of pine xylophilus inhibitors, the steps are as follows: 1) collection of pine xylophilus, that is, isolation of pine xylophilus from infected wood, and then expanded cultivation; 2) screening of pine xylophilus, to be cultivated Nematodes are distinguished and counted according to adults and larvae; 3) Test plate production, that is, control and test areas are set in the microplate and a certain concentration of reagents are configured; 4) Data recording and processing, that is, the change of life and death of nematodes over time is recorded , statistical drug efficacy, screening.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for high-throughput screening of pine wood nematode inhibitors. Background technique [0002] Pine xylophilus disease is a major international quarantine disease caused by pine wilting and death caused by pine xylophilus. Since the first discovery of pine wood nematode disease in Zijinshan black pine in Nanjing in 1982, the disease has occurred in more than 300 cities and counties in 18 provinces. Over the past 30 years, pine wood nematode has been the number one forest disease and insect pest in my country. Pine trees are the most important tree species for afforestation in my country. The national pine forest covers an area of ​​more than 500 million mu. The main tree species are Masson pine, black pine, red pine, Ryukyu pine, Huashan pine, Yunnan pine, Simao pine, Huangshan pine, Huashan pine, red pine, etc. All are susceptible species. In terms of controlling pine wood...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/15
CPCG01N33/15
Inventor 彭华正金群英朱汤军张飞英叶华琳戚全英
Owner ZHEJIANG FORESTRY ACAD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com