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

Application of specific target primer in simultaneous and rapid identification of two pathogenic bacteria infected by strawberry

A technology for pathogenic bacteria, strawberry anthracnose, applied in the biological field, can solve the problems of single LAMP detection, lack of anthrax LAMP detection, detection of strawberry Fusarium oxysporum and anthracnose and other problems

Pending Publication Date: 2020-10-30
新疆生产建设兵团第十二师农业科学研究所
View PDF1 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The application of this technology in the detection of strawberry virus disease at home and abroad is limited to the detection of a single strain of strawberry anthracnose, lack of LAMP detection of anthracnose, and most of them are single LAMP detection. At present, there is no use of LAMP detection to detect strawberry at the same time Reports on Two Pathogenic Bacteria of Fusarium oxysporum and Bacillus anthracnose

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
  • Application of specific target primer in simultaneous and rapid identification of two pathogenic bacteria infected by strawberry
  • Application of specific target primer in simultaneous and rapid identification of two pathogenic bacteria infected by strawberry
  • Application of specific target primer in simultaneous and rapid identification of two pathogenic bacteria infected by strawberry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1: The LAMP primer sequence of Fusarium oxysporum IGS gene and Fragaria anthracis ITS gene

[0043] The application of specific target primers in the rapid identification of two pathogenic bacteria infected with strawberries at the same time. The specific target primers are composed of the LAMP primer sequences of the IGS gene of Fusarium oxysporum and the ITS gene of B. fragariae. The LAMP detection of the IGS gene of F. Primers FIP, BIP, F3, B3, LoopF, LoopB, specifically as shown in SEQID NO.6-11.

Embodiment 2

[0044] Example 2: A LAMP detection kit for simultaneous and rapid identification of two pathogenic bacteria infected with strawberry, Fusarium oxysporum and Anthrax

[0045] A LAMP detection kit for rapid identification of two pathogens of strawberry infection Fusarium oxysporum and anthracnose at the same time, the detection kit includes 1.0 μL of Bst DNA polymerase (8U), 2.5 μL of 10×Bst Reaction Buffer, 4 μL of 0.8 mol / L betaine, 3.5 μL of 1.4 mmol / L dNTPs, 4 μL of 8 mmol / L MgSO 4 oxysporum LAMP primer 20 μmol / L F3 and B3 primer 0.25 μL each, 40 μmol / L FIP and BIP primer 1 μL each, 40 μmol / L loop primer Loop 0.5 μL; anthracis LAMP primer 10 μmol / L F3 and B3 0.25 μL of each primer, 1 μL each of 20 μmol / L FIP and BIP primers, 1 μL each of 10 μmol / L loop primer Loop, 1 μL DNA template (10 ng / μL), add double distilled water to make the total volume of the reaction system reach 25 μL.

Embodiment 3

[0046] Example 3: Application of a LAMP detection kit for simultaneous rapid identification of strawberry infection of two pathogenic bacteria, Fusarium oxysporum and Anthrax

[0047] Specifically, the present invention provides an application of a LAMP detection kit for quickly identifying two pathogenic bacteria of strawberry infection Fusarium oxysporum and Anthrax at the same time. The specific application technical scheme includes the following steps:

[0048] (1) Primer design and synthesis

[0049] According to the IGS gene of Fusarium oxysporum known in GenBank, the second half of the ITS gene of Anthracis was used as the target sequence, which was non-repetitive sequence with the ITS region of Fusarium, and two sets of specific primers were designed using the online PrimerExplorerVer.5 primer design software. LAMP primers:

[0050] The sequence of LAMP primers for specifically detecting the IGS gene of Fusarium oxysporum is:

[0051] FIP: GCAACCTGGCGAGGACGAAACACTGTC...

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 application of a specific target primer in simultaneous and rapid identification of two pathogenic bacteria infected by strawberry. The specific target primer is composed of LAMP primer sequences of a fusarium oxysporum IGS gene and a strawberry colletotrichum ITS gene. The method comprises the following steps: designing an LAMP primer of a specific ITS sequence of fusariumoxysporum, an LAMP primer of a CYP sequence, an LAMP primer of an IGS sequence and other technical means, and finally selecting the IGS sequence to design the LAMP primer for identifying strawberry fusarium oxysporum. Meanwhile, the characteristic of extremely low homology between the rear half part of the colletotrichum strain ITS and other genera is utilized, an LAMP specific primer of an ITS sequence is designed; meanwhile, an ITS primer and an IGS primer are added, fusarium oxysporum and colletotrichum gloeosporioides in strawberries are rapidly and accurately distinguished and detected for the first time through the LAMP technology, the detection sensitivity is improved by 1000 times compared with that of a PCR technology, the price is low, the detection time is short, the method issimple, and the LAMP technology plays an important role in real-time monitoring and early warning of strawberry pathogenic bacteria.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the technical field of the application of a unique target primer in simultaneous rapid detection of two pathogenic bacteria, Fusarium oxysporum and Bacillus anthracis. Background technique [0002] Strawberry (Fragaria ananassa Duch.) belongs to the Rosaceae Strawberry genus perennial herbaceous plant, the pulp is delicious and juicy, and it can be called the "Queen of Fruits". In recent years, the planting area of ​​strawberry has been increasing year by year, and the varieties are constantly being updated, with frequent replanting, introduction and continuous cropping. Among them, strawberry wilt and anthracnose are the two most common and currently most important diseases. Strawberry wilt is a systemic disease caused by Fusarium oxysporum infection from the roots, and it can occur from the seedling stage to the harvest stage. After the strawberry is infected with straw...

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): C12Q1/6895C12Q1/689C12Q1/6844C12Q1/04C12R1/77C12R1/07
CPCC12Q1/6895C12Q1/689C12Q1/6844C12Q2531/119C12Q2563/107
Inventor 史芳芳王雷张延磊梁武军陈烨华
Owner 新疆生产建设兵团第十二师农业科学研究所
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