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

Molecular identification method for two kinds of leafhopper egg parasitic wasps in tea garden

A molecular identification and parasitic wasp technology, applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of insufficient control of leafhopper population, difficulty in determining the morphological identification of parasitic wasp species, etc., and achieve accurate and rapid identification. Effect

Inactive Publication Date: 2016-11-09
TEA RES INST CHINESE ACAD OF AGRI SCI
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the spider's habits and feeding habits, its control over the leafhopper population is less than 20%
[0005] The tea leafhopper has a small body, and it usually lays eggs on the tender stems of the 2nd to 4th leaves under the buds. The eggs are very small, and its egg parasitoids must rely on a microscope to see them. Therefore, the determination of the parasitoid species depends only on Morphological identification is more difficult

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
  • Molecular identification method for two kinds of leafhopper egg parasitic wasps in tea garden
  • Molecular identification method for two kinds of leafhopper egg parasitic wasps in tea garden
  • Molecular identification method for two kinds of leafhopper egg parasitic wasps in tea garden

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Collection of parasitoid eggs of tea leafhopper eggs

[0025] The high incidence period of the small green leafhopper in tea gardens in Zhejiang Province is from June to October, and its egg parasitoids have an obvious follow-up effect on leafhoppers, so the collection of parasitoids should follow the two peak periods of leafhoppers. Every few days in the tea garden, randomly cut one-bud five-leaf branches with obvious puncture marks on the tender stems, and bring them back to the laboratory and put them in water for cultivation. During this period, the epidermis of tea shoots and tender stems was girdled, and the eggs of small green leafhoppers were picked out and placed on filter paper soaked in distilled water for moisture observation. After the parasitoids emerged, the parasitoid eggs of leafhoppers were collected with glass finger tubes. The eclosion parasitoids were first photographed with a three-dimensional depth-of-field system for preliminary morphol...

Embodiment 2

[0026] The extraction of embodiment 2 genome DNA

[0027] Air-dry the preserved single parasitic wasp body of the green leafhopper, add 0.5 ml lysis buffer (100 mM Tris-HCl, 5 mM EDTA, 0.5 % SDS, 200 mM NaCl, pH 8.0) and 1 steel ball, shake and crush, Add 5 ml of proteinase K (20 mg / ml) and digest in a water bath at 55°C for 5 hours. Use an equal volume of phenol:chloroform:isoamyl alcohol (25:24:1) and chloroform:isoamyl alcohol (24:1) to extract once, centrifuge at 12,000 r / min for 10 min at 4°C, and use the supernatant Add 2 times the volume of absolute ethanol and add 1 / 10 volume of NaAc (3 mol / L), precipitate at -20°C for 30 min, centrifuge at 12,000 r / min at 4°C for 10 min, wash the DNA pellet with 70% ethanol, and ultra-clean Add 100 ml of sterile deionized water after the workbench is air-dried, and store in a -20°C refrigerator.

Embodiment 3

[0028] Example 3 Molecular Identification Marker Gene Screening

[0029] A series of primers including 28S, 18S, COI and 16sRNA were designed and synthesized according to the marker genes used in the study of the evolution of Hymenoptera insects. Using parasitoid DNA as a template, amplify each gene under the same conditions, and use a 20ml system for PCR reaction (template 1ml, 10×buffer 2.0ml, dNTP 1.6ml, forward and reverse primers 0.5ml, Taq enzyme 0.2ml, plus sterilized Double distilled water to 20ml), the PCR amplification procedure is as follows: first, pre-denature at 94°C for 3 minutes; then perform 35 cycles of 94°C for 30 s, 55°C for 30 s, and 72°C for 1 min; finally, extend at 72°C for 10 min. Take 10ml of the PCR product and test it by 1% agarose gel electrophoresis (for the test results, see figure 2 ). A total of 9 pairs of primer combinations produced products, which were sequenced to remove repetitive sequences. Considering their representativeness in the s...

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 provides a molecular identification method for two kinds of empoasca flavescens advantageous parasitic wasps in a tea garden. Compared with a traditional morphological identification method, the method has the advantages that professional equipment (such as a three-dimensional depth-of-field microscopic photographing system) is not needed; a strict morphological taxonomy background is not required; the method is accurate and fast and easy to operate and can be applied to molecular identification of other kinds of egg parasitic wasps. The method is characterized by comprising the following steps of 1 leafhopper spawning branch collection, 2 leafhopper egg parasitic wasp collection, 3 morphological photograph acquisition, 4 parasitic wasp DNA extraction, 5 gene clone and analysis and 6 molecular systematic identification.

Description

technical field [0001] The invention belongs to the field of tea garden insect evolution, and in particular relates to a molecular identification method for two kinds of tea garden leafhopper parasitoids. Background technique [0002] The tea industry is one of my country's traditional characteristic agricultural industries. About 80 million people in the country are engaged in tea production and sales. Tea is not only the main source of income for farmers in the southern mountainous and mid-mountainous areas of my country, but also plays a very important role in the adjustment of agricultural industrial structure, the construction of new rural areas and the earning of foreign exchange through export. During the growth and development of tea trees, they often suffer from various diseases and insect pests, especially the bud and leaf parts have the most types of diseases and insect pests, which are the most harmful. Leafhopper is currently the most important piercing-sucking...

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 Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888
Inventor 付建玉周孝贵
Owner TEA RES INST CHINESE ACAD OF AGRI SCI
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