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

Method for rapidly detecting impatiens necrotic spot virus from oncidium

A detection method and technology of oncidium, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of small transmission medium, harm to orchid industry, fast transmission and reproduction speed, etc. The effect of improved sex and specificity, high sensitivity and good specificity

Inactive Publication Date: 2008-11-19
CHECKOUT & QUARANTINE TECH CENT YUNNAN ENTRY &EXIT CHECKOUT & QUARANTINE BUR
View PDF0 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, impatiens necrotic spot virus is easy to cause serious harm to the orchid industry due to its wide range of hosts, small transmission vectors, and fast transmission and reproduction speeds. This is a technical problem that needs to be solved urgently
No related reports have been found in the prior art

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
  • Method for rapidly detecting impatiens necrotic spot virus from oncidium
  • Method for rapidly detecting impatiens necrotic spot virus from oncidium
  • Method for rapidly detecting impatiens necrotic spot virus from oncidium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] ——RT-PCR detection

[0022] Samples were taken from the symptomatic part of the susceptible plant, the leaf tip far away from the symptom, and the healthy plant, and the total plant RNA was extracted. After synthesizing cDNA, conventional PCR amplification was performed using ZI2F and ZI2R as specific primers. PCR reaction system 25 μL: 2 μL of the above reverse transcription product, 3 μL of 10×PCR Buffer, Mg 2+ (25mM) 1.2μL, dNTP Mixture (2.5mM each) 0.4μL, ZI2F 0.4μL, ZI2R 0.4μL, Taq enzyme (5U / μL) 0.2μL and sterile water 17.4μL. Reaction program: 94°C for 5min; 30 cycles of 94°C for 30s, 45°C for 30s, and 72°C for 40s; 72°C for 8min.

[0023] RT-PCR amplification products were detected by 1% agarose gel electrophoresis, and the detection results were as follows: figure 2 , after PCR amplification on the leaves showing symptoms, a target band consistent with the expected fragment size can be seen at 500bp (see figure 2 , lanes 1, 2). Target bands with the expec...

Embodiment 2

[0026] ——Nested PCR detection

[0027] The first round of PCR amplification was performed with ZI1F and ZI1R as outer primers, and the second round of amplification was performed with ZI1F and ZI3R, ZI3F and ZI1R, ZI3F and ZI3R as inner primers respectively. Template 2μL, 10×PCR Buffer 3μL, Mg 2+ (25mM) 1.2μL, dNTP Mixture (2.5mM each) 0.4μL, upstream primer 0.4μL, downstream primer 0.4μL, Taq enzyme (5U / μL) 0.2μL and sterilized water 17.4μL. Reaction program: 94°C for 5min; 30 cycles of 94°C for 30s, 52°C for 30s, 72°C for 40s; 72°C for 8min.

[0028] At the same time, the cDNA was multiplied by 10 times, 10 -2 times, 10 -3 times, 10 -4 times, 10 -5 After two-fold dilution, primers ZI1F and ZI1R were used for the first round of amplification, and then ZI3F and ZI3R were used as primers for the second round of amplification to verify the sensitivity of nested PCR.

[0029] 1% agarose gel electrophoresis detection results showed that ZI1F and ZI3R amplified fragments 850b...

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 discloses a method for quickly detecting Impatiens necrotic spot virus on oncidiums. The method uses Impatiens necrotic spot virus antiserum to detect the Impatiens necrotic spot virus on oncidium leaves which represent a concentric circle chlorosis spot symptom; a special primer ZI2R of the Impatiens necrotic spot virus is utilized to synthesize cDNA; the synthesized cDNA is applied, and a special primer ZI2F / ZI2R and a special primer ZI1F / ZI1R are respectively used to amplify to establish a RT-PCR assay. The method can also perform secondary amplification to a PCR product of ZI1F / ZI1R amplification respectively with ZI1F / ZI3R, ZI3F / ZI1R and ZI3F / ZI3R as inner primers to establish a nested PCR assay. The method has high sensitivity and good specificity, and provides an effective detection means for timely preventing and curing the attacking of the Impatiens necrotic spot virus to strains.

Description

technical field [0001] The invention belongs to the technical field of plant virus detection, and more specifically, the invention relates to a method for detecting impatiens necrotic spot virus of oncidium seedlings. Background technique [0002] Oncidium, also known as dancing orchid and golden saucer orchid, is a perennial herbaceous flower native to Central and South America and southern North America. It has become one of the most important orchid varieties in the world due to its bright colors and rich flower patterns. It is now cultivated all over the world. With the expansion of orchid planting range and the application of asexual reproduction technology, the occurrence of viral diseases on orchids is also becoming more and more serious. It has been reported that 27 virus diseases infect orchids, among which the common virus diseases on oncidium are Cymbidium mosaic virus (CyMV) and Odontoglossum ringspot virus (ORSV). Current research focuses on the detection of ...

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): C12Q1/70C12Q1/68
Inventor 丁元明张巧萍刘毅周剑段禄华和万忠和捷
Owner CHECKOUT & QUARANTINE TECH CENT YUNNAN ENTRY &EXIT CHECKOUT & QUARANTINE BUR
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