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

Wheat yellow streak virus and its nash detection method and detection kit

A stripe virus and detection method technology, applied in the field of agricultural biology, can solve problems such as difficult promotion by grass-roots plant protection departments, difficulty in mastering experimental techniques, and inability to complete accurate identification, achieving good application prospects, taking into account practicability and rapidity, and saving energy. The effect of testing costs

Inactive Publication Date: 2021-06-25
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But as far as the new virus is concerned, because it and WDV are both transmitted by the leafhopper and have similar symptoms, biological identification cannot complete accurate identification; serology has the advantages of fast, simple, high-throughput, etc. Antiserum-dependent virus detection technology is the most mature and widely used ELISA technology
However, due to the limitation of antiserum preparation level, there is no commercial antiserum in China; molecular detection technology is widely used in the rapid detection and identification of plant viruses and the early screening of diseases due to its advantages of rapidity, sensitivity, specificity and accuracy. checking and diagnosing
PCR method is commonly used, but these experimental techniques are not easy to master, and are affected by cost issues such as expensive reagents and instruments, so it is difficult to promote in grassroots plant protection departments

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
  • Wheat yellow streak virus and its nash detection method and detection kit
  • Wheat yellow streak virus and its nash detection method and detection kit
  • Wheat yellow streak virus and its nash detection method and detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1. The preparation of the medium leafhopper and WYSV toxin source samples

[0047] 1.1 The source and preservation of the medium leafhopper

[0048] The different sand leafhoppers are collected from ordinary fields in Hancheng, Shaanxi, and are reared on healthy wheat after detoxification and purification. The feeding conditions are: 22±1°C, 20000lx light incubator.

[0049] 1.2 Source and preservation of wheat yellow streak virus (WYSV)

[0050] In April 2016, a new disease symptom was discovered during the field investigation of the disease in Hancheng, Shaanxi. It was different from the symptoms caused by the wheat dwarf virus disease. The leaves become chlorotic along the veins, gradually develop from the tips of the leaves to dry, and finally die. 2,000 leafhoppers were collected from diseased fields and stored in insect-proof net cages in the greenhouse. After the insects were inoculated on Yangmai 12 at the one-leaf stage, they were transferred to a li...

Embodiment 2

[0052] Example 2. Identification of Wheat Yellow Streak Virus (WYSV)

[0053] 2.1 Biological identification

[0054] In order to detect the specificity of virus transmission by mediators, three species of non-toxic brown planthopper (Nilaparvata lugens), white-backed planthopper (Laodelphax striatellus) and white-backed planthopper (Sogatella furcifera Horváth) and wheat long tube aphid (Sitobion avenae) were selected. ), wheat aphid (Schizaphis gramienum) and cereal aphid (Rhopalosiphum padi) were used as experimental insects for virus transmission experiments. In addition, barley (variety name: Longpimai No. 3) and oats (variety name: Anhei oat) were selected for preliminary host range identification. The feeding and transmission steps of the above experiment are the same as those in Example 1.2. All the tested plants were placed in the light incubator to grow, and the onset symptoms were observed after 2-3 weeks. The results of biological experiments show that WYSV can be...

Embodiment 3

[0067] Example 3. Extraction of plant total RNA

[0068] The total RNA of the wheat samples to be tested was extracted by TRIzol (Invitrogen, USA) method. The specific extraction process is as follows: put 0.1g of fresh or frozen sample into a 1.5mL centrifuge tube, quickly transfer to liquid nitrogen to freeze, fully grind to powder, then add 1mL of TRIzol, shake vigorously to mix, and stand on ice for 5min ; Add 200 μL chloroform, shake vigorously for 15 seconds, and let stand on ice for 5 minutes. Centrifuge at 12,000rpm at 4°C for 10min; transfer the supernatant to a new centrifuge tube, add isopropanol equal to the volume of the supernatant, mix gently by inversion, and let stand on ice for 10min; centrifuge at 12,000rpm at 4°C for 15min, remove Supernatant; add 1 mL of pre-cooled 75% ethanol (prepared with sterilized DEPC water) to wash the precipitate. Centrifuge at 12,000rpm at 4°C for 5 minutes, discard the supernatant, and repeat the above washing steps to thorough...

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 a wheat yellow streak virus and a NASH detection method and detection kit thereof. The present invention discovers a new virus, namely wheat yellow streak virus WYSV, the full-length sequence of which is shown in SEQ ID NO. , using digoxigenin as a marker to prepare DNA probes to detect WYSV virus. The invention also provides a WYSV virus kit for NASH detection, the kit includes: a probe PCR reaction solution, a NASH reaction reagent, a nitrocellulose membrane, a hybridization bag, a WYSV positive control sample and a negative control sample. The detection method and detection kit can detect samples in batches at one time, can detect WYSV virus in a short time with low cost and high efficiency, and have reliable detection results, easy operation, strong specificity, high sensitivity, short time consumption, and good application prospect.

Description

technical field [0001] The invention belongs to the technical field of agricultural biology, and in particular relates to a detection method and a detection kit for wheat yellow streak virus (Wheatyellowstriate virus, WYSV) and NASH thereof, which can achieve the purpose of fast, sensitive and convenient detection of WYSV in plants . Background technique [0002] Wheat is one of the major food crops around the world. Viral diseases are an important class of diseases that endanger wheat production, and more than 50 viral pathogens have been reported. The virus can cause serious symptoms such as leaf chlorosis, plant dwarfing, and effective tiller reduction in infected wheat plants, which have a huge impact on wheat growth and yield. Many viruses that infect wheat are transmitted specifically by vector insects, including aphids, SBPH, and leafhoppers. With global warming, no-tillage and prohibition of highly toxic pesticides and other factors, the population of virus-transm...

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): C12N15/40C12Q1/70C12Q1/6811C12Q1/6834C12Q1/6804C12N15/11C12R1/94
CPCC07K14/005C12N2760/00021C12Q1/6804C12Q1/6811C12Q1/6834C12Q1/701C12Q2531/113C12Q2565/125C12Q2563/131
Inventor 刘艳杜真真王锡锋
Owner INST OF PLANT PROTECTION 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