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

Detection of Phytophthora cedarii Root Rot Pathogen by Loop-Mediated Isothermal Amplification (Lamp) Technology Based on Color Judgment

A technology for root rot fungus and Phytophthora is applied in the field of molecular detection for detecting Phytophthora cedar root rot fungus and LAMP primers for detecting Phytophthora cedar root rot fungus. , poor specificity and other problems, to achieve the effect of expanding the scope of use, time-consuming, and high accuracy

Inactive Publication Date: 2020-06-19
中华人民共和国昆山出入境检验检疫局
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0016] The technical problem to be solved by the present invention is to solve the problems that the biological detection method of Phytophthora cedarii root rot in the prior art requires a long cycle, time-consuming, laborious, cumbersome, poor specificity and PCR detection technology needs a thermal cycle instrument, high cost, The problem of being unable to quickly detect Phytophthora cedarum root rot, thus providing a new molecular detection method for Phytophthora cedarum root rot, LAMP detection of Phytophthora cedarum root rot, the detection cycle is short (only 80min), specificity Strong performance, high sensitivity, the test results can be observed with naked eyes without the need for expensive instruments, low cost, and easy to operate

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
  • Detection of Phytophthora cedarii Root Rot Pathogen by Loop-Mediated Isothermal Amplification (Lamp) Technology Based on Color Judgment
  • Detection of Phytophthora cedarii Root Rot Pathogen by Loop-Mediated Isothermal Amplification (Lamp) Technology Based on Color Judgment
  • Detection of Phytophthora cedarii Root Rot Pathogen by Loop-Mediated Isothermal Amplification (Lamp) Technology Based on Color Judgment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Selecting novel target genes, designing and screening LAMP primers for detecting Phytophthora cedarii root rot, and establishing a LAMP detection system

[0042] (1) Selection of novel target genes and design and screening of primers

[0043] The selection of new target genes and the screening of primers are the key factors for LAMP detection. At present, molecular detection is widely used based on the ribosomal transcriptional spacer (ITS) target, but because the ITS sequence does not have enough variation sites to distinguish all Phytophthora species, it is necessary to discover new molecular detection targets. First, we searched relevant literature and selected available targets such as tRNA (transfer RNA) EF1α (translation elongation factor), Ypt1 (guanosine triphosphate-binding protein gene), and beta tubulin gene. Taking the target Ypt1 as an example, download the Ypt1 gene sequence of Phytophthora cedarii root rot and the Ypt1 gene sequences of other ...

Embodiment 2

[0054] Embodiment 2: Preparation of DNA template

[0055] The DNA extracted from the sample is used as a template for the LAMP reaction. The specific process is as follows:

[0056] (1) Bacterial strain culture and preparation of mycelium powder

[0057] The Phytophthora strains for testing are transferred to LBA (Lima bean medium (Zheng Xiaobo. Phytophthora and its research technology. China Agricultural Press. 1997)) plate, and other bacterial strains are transferred to PDA (Potato Dextrose Solid Medium (Erwin , D.C.; Ribeiro, O.K. Phytophthora lateralis. In: Phytophthora disease worldwide. American Phytopathological Society, St.Paul (US), 1996, pp 365-367)) plate, 25 ℃ dark culture for 3 days and cut 10 pieces from the edge of the colony 2× 2mm hyphae block, the bacterial strain of Phytophthora is transferred to V8 liquid medium (Zheng Xiaobo. Phytophthora and its research technology. China Agricultural Press. 1997), other bacterial strains are transferred to PDB (Potato G...

Embodiment 3

[0060] Example 3: Detection of specificity and sensitivity of LAMP primers

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

PropertyMeasurementUnit
diameteraaaaaaaaaa
diameteraaaaaaaaaa
Login to View More

Abstract

The invention discloses a molecular detection method for detecting phytophthora sojae of cedar by the color determination-based loop-mediated isothermal amplification (LAMP) technology and a primer thereof. The primer sequence is as shown respectively in SEQ ID NO.1 to SEQ ID NO.5. The detection system can be used for quickly, conveniently, efficiently, specifically and sensitively detecting the phytophthora sojae of cedar under a 62 DEG C isothermal condition, a complicated instrument is not required, a new technical platform is provided to detection of phytophthora sojae of cedar, and field detection on phytophthora sojae of cedar can be well fulfilled. The method is applicable to inspection and quarantine of imported and exported plants and plant products and investigation, quick diagnosis, detection and the like of diseases, and has an important meaning for preventing phytophthora sojae of cedar from being introduced into China. Furthermore, the establishment of the system provides technical guidance and theoretic base to detection of other pathogenic bacteria.

Description

technical field [0001] The invention relates to a LAMP primer for detecting Phytophthora lateralis Tucker et Mibrath and a molecular detection method for detecting Phytophthora lateralis Tucker et Mibrath, belonging to the field of biotechnology. Background technique [0002] Phytophthora cedarum root rot is an important destructive plant pathogen that can infect American cypress Chamaecyparis lawsoniana (A. Murray bis) Parlatore and cause severe root rot. The disease was first reported on ornamental plants near Seattle and Washington, USA in 1923. Until 1942, the pathogen was discovered and named in Willamette Valley, Oregon, USA. The serious economic losses caused by the horticultural cultivars almost completely destroyed the valuable horticultural cultivars, and then the bacteria were introduced into Canada and France, which had a serious impact on the local ornamental plants and forestry trade. [1] . Due to the serious damage and rapid spread of the disease, and the ex...

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): C12Q1/6895C12Q1/6844C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6844C12Q1/6895C12Q2531/119
Inventor 纪睿曾丹丹廖太林郑小波张正光周锐吴军
Owner 中华人民共和国昆山出入境检验检疫局
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