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

Molecular markers and detection kits related to drug resistance of shepherd's purse

A molecular marker and drug resistance technology, which is used in the determination/examination of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc.

Active Publication Date: 2016-07-13
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

In the research report, only one ALS gene of shepherd's purse was cloned, but we found in the study that shepherd's purse has two ALS genes, named JC-ALS-1 and JC-ALS-2, both of which have functions, and any A gene mutation in a conserved region can cause drug resistance

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
  • Molecular markers and detection kits related to drug resistance of shepherd's purse
  • Molecular markers and detection kits related to drug resistance of shepherd's purse
  • Molecular markers and detection kits related to drug resistance of shepherd's purse

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1 is used for detecting the synthesis of the PCR primer of anti-ALS inhibitor class herbicide shepherd's purse

[0038] Query the shepherd's purse ALS gene sequence (GenBank accession number: HQ880660.1) in the GenBank database, according to the sequence information found, use primer design biology software, design for specific amplification JC-ALS-1 and JC-ALS- The PCR primers of the conserved region sequences of 2 were used, and the sequences at both ends were obtained by 3'RACE and chromosome walking methods, and finally the two ALS gene sequences of shepherd's purse were obtained respectively.

[0039] The primer sequences are as follows:

[0040] Forward primer JC-ALS-F: 5'-TATTCGCTTACCCAGGTG-3';

[0041] Reverse primer JC-ALS-R: 5'-GGTTCTGAGTTTCATCTCTCA-3'.

[0042] Primer synthesis was completed by Beijing Liuhe Huada Gene Technology Co., Ltd.

Embodiment 2

[0043] The PCR detection method of embodiment 2 anti-ALS inhibitor class herbicide shepherd's purse

[0044] Experimental material: Shepherd's purse (Capsellabursa-pastoris) collected from different regions.

[0045] experimental method:

[0046] 1. Preparation of shepherd's purse DNA

[0047] Take 200 mg of fresh leaves of shepherd's purse, grind with liquid nitrogen, and extract the DNA of each shepherd's purse leaf by conventional CTAB method.

[0048] 2. Specific primers for detecting mutation sites of shepherd's purse JC-ALS-1 and JC-ALS-2, see Example 1 for the primer sequences.

[0049] 3. PCR reaction system for detecting ALS mutation site of shepherd's purse

[0050] PCR reaction system, in which 10×PCR reaction buffer 2μL, 15mM MgCl 2 0.5 μL, 1 μL of 2.5 mM dNTPs, 0.5 μL of each 10 μM primer, 1 U of TaqDNA polymerase, 1 μL of DNA template, and the rest in sterile double-distilled water, with a final volume of 20 μL.

[0051] 4. PCR amplification program for dete...

Embodiment 3

[0060] Embodiment 3PCR method detects the shepherd's purse of anti-ALS inhibitor in the field

[0061] The detection method is as follows:

[0062] 1. Sample collection and DNA preparation

[0063] In order to verify the feasibility of the PCR detection method, samples were collected from farmland where drug-resistant shepherd's purse was suspected to have occurred for two consecutive years in 2013 and 2014, and tested in the laboratory. Tribesulfuron-methyl was applied once to control shepherd's purse in these fields after the wheat turned green, but it was ineffective, so it was suspected that there might be resistance to tribenuron-methyl. Therefore, the shepherd's purse plants were collected from these suspected drug-resistant farmlands, and after proper moisturizing treatment, they were mailed to the laboratory for testing. Take 200 mg of fresh leaves of shepherd's purse, grind with liquid nitrogen, and extract the DNA of each shepherd's purse leaf by conventional CTAB ...

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
Login to View More

Abstract

The invention provides a shepherd's purse drug resistance related molecular marker and a specific PCR (polymerase chain reaction) primer pair for detecting anti-acetolactate synthase inhibitor weedicide shepherd's purse. The PCR primer pair comprises a primer pair for specifically amplifying shepherd's purse JC-ALS gene. The PCR detection method by using the primers has excellent specificity and sensitivity, can implement quick identification of suspected anti-ALS inhibitor shepherd's purse in the field and conformation of mutant sites, thereby instructing the scientific treatment on resistant weed in production practice.

Description

technical field [0001] The invention relates to a detection technology for herbicide-resistant weeds, in particular to a molecular marker related to drug resistance of shepherd's purse and a detection kit thereof. Background technique [0002] Acetolactate synthase (ALS for short), also known as acetohydroxyacid synthase (AHAS for short), is the key to inducing the biosynthesis of valine, leucine and isoleucine in plants and microorganisms. The enzyme is also an important herbicide target. The development and use of ALS inhibitor herbicides began in the early 1980s. Due to the obvious advantages of such herbicides, such as super high activity, low toxicity to humans and animals, and environmental friendliness, they have attracted widespread attention. At present, 50 Multiple varieties were developed and used. Since the 1980s, my country has successively promoted the use of tribenuron-methyl, metsulfuron-methyl, chlorsulfuron, florasulam, acesulfame, etc. in winter wheat fi...

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): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 崔海兰李香菊张宏军
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