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

PCR testing and identification optimizing process for carrot black rot

An optimization method, carrot technology, applied in the field of crop disease prevention and control, can solve the problem of undetectable products, etc., and achieve the effect of improving amplification efficiency and sensitivity

Inactive Publication Date: 2006-08-30
YUNNAN AGRICULTURAL UNIVERSITY
View PDF0 Cites 14 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing PCR programs do not have pre-denaturation, resulting in often undetectable products after amplification

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
  • PCR testing and identification optimizing process for carrot black rot

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1 PCR amplification results of two primers ARF2 / ARR3 isolated from carrot disease samples (one is a strain of Fusarium and the other is a strain of carrot black rot) using this optimization program

[0030] Specific steps are as follows:

[0031] 1) Isolation of carrot black rot fungus strains

[0032] Carrot disease samples were taken at the junction of disease and health 1 × 0.5cm, soaked in 70% alcohol for 30 seconds, 0.1% mercuric chloride for 3 minutes, rinsed three times with sterilized water, each time for 3 minutes, dried in PDA (potato-dextrose -agar) culture medium at 25°C for 7 days.

[0033] 2) Genomic DNA extraction of carrot black rot pathogen was carried out by CTAB method

[0034] 3) Specific primers for detecting carrot black rot

[0035] Primer: ARF2 5-AAT CAG CGT CAG TAA ACA AAC G-3

[0036] ARR3 5-AGA GGC TTT GTG GAT GCT G-3

[0037] Primers were diluted to 10um, aliquoted and stored at -20°C. Primers ARF2 / ARR3 specifically amplified a 34...

Embodiment 2

[0051] Using the specific primers ADF2 / ADR1 of Alternaria dauci to perform PCR amplification with optimized and non-optimized procedures using the genomic DNA of Carrot black rot fungus strain as template

[0052] Method is the same as example 1, just adopt the specific primer with Alternaria dauci

[0053] ADF2: 5-GCA ATC AGC GTC AGT AAC AAC A-3

[0054] ADR1: 5-CGC AAG GGG AGA CAA AAA-3

[0055] Primers were diluted to 10um, aliquoted and stored at -20°C. (Synthesized by Beijing Sanbo Yuanzhi Co., Ltd.) Implementation results:

[0056] Using the specific primer ADF2 / ADR1 of Alternaria dauci to carry out optimized and non-optimized PCR amplification with the genomic DNA of Carrot black rot fungus strain as template, no band was amplified. It shows that the carrot black rot in my country is caused by the fungus Alternaria radicina.

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

This invention is PCR method used to detect and determine carrot black rot germ. It is propitious to detect carrot black rot and earlier diagnoses, and it can exactly determine the Alternaria radicina of the disease. It belongs to crop disease protection field. This method detects the germ according to the special primer brought by the germ, the amplifying product is 343bp. At the same time, the amplification program of the PCR is optimized by this method, 94 degree centigrade pre-changing 4min is added, and the sensibility is improved. It is the supplement and improvement of the method of detecting carrot black rot germ special prime.

Description

technical field [0001] The invention relates to a method for PCR detection and identification optimization of carrot black rot, belonging to the field of crop disease prevention and control. Background technique [0002] my country is a big producer of high-quality carrot raw materials. Heilongjiang, Jilin, Qinghai, Shaanxi, Inner Mongolia, Xinjiang, Gansu, Ningxia and other provinces are the main production areas of high-quality carrot raw materials in my country. The combination of resource advantages and technological advantages will make my country the world's largest carrot high-quality industrial raw materials and carrots. Processing and exporting countries of functional foods. In recent years, the area of ​​carrot planting has expanded rapidly in various places. Due to factors such as poor production management, carrot diseases have gradually increased. [0003] The fungi Alternaria radicina and A.dauci are the most important seed-borne and soil-borne fungi on carrot...

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
IPC IPC(8): C12Q1/68
Inventor 黄琼赵莹莹王云月孙跃先焦玉霞
Owner YUNNAN AGRICULTURAL UNIVERSITY
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