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

Primers and methods for identifying Rhododendron xing'an and Rhododendron japonica

A technique for rhododendron and its products, which is applied in the field of primers for identifying Rhododendron xing'an and Rhododendron chinensis, can solve the problems of identification and the inability to separate the two species

Active Publication Date: 2022-04-01
NORTHEAST NORMAL UNIVERSITY
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the recent phylogenetic tree based on chloroplasts cannot separate the two species. There are shared haplotypes between the two species, and the distribution overlap of the two species in the Jilin area, so the identification of the two species caused trouble

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
  • Primers and methods for identifying Rhododendron xing'an and Rhododendron japonica
  • Primers and methods for identifying Rhododendron xing'an and Rhododendron japonica
  • Primers and methods for identifying Rhododendron xing'an and Rhododendron japonica

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Design of primers

[0028] Based on the previously sequenced SLAF data (PRJNA589346) of Rhododendron xing'an and Rhododendron japonica, and based on the reference genome Rhododendron lantana (R. delavayi), all reads of all samples were aligned and clustered using BLAT software to obtain the SLAF tag sequence. Afterwards, use MISA-web (http: / / misaweb.ipk-gatersleben.de / ) to scan these polymorphic SLAF sequences for microsatellite sites, and set the repeating unit as a microsatellite site of two bases The minimum number of repeats is 6, and the minimum repeats of three, four, five and six bases for the rest of the repeating units are all set to 5 times. Two microsatellite loci are regarded as a microsatellite locus if the distance between them is less than 100bp.

[0029] At the same time, the software SAMtools and GATK were used to identify the insertion / deletion sites of the population, and the parameters of the software were the default parameters. The obta...

Embodiment 2

[0034] Embodiment 2SSR-PCR and product detection

[0035] (1) Use the 4 × CTAB method to extract the whole genome DNA, and use 1% agarose gel for detection to ensure that the concentration reaches 20ng / ul or more, and the DNA that passes the test is stored at -20 ℃;

[0036] (2) Use 20 μL fluorescent PCR system for population PCR amplification: 50 ng of the above genomic DNA, 1 × PCRbuffer (plus Mg2+), 0.2 mMdNTP and 0.5 μM each of the forward and reverse primers shown in Table 1, and the forward primers use different Fluorescent markers (FAM, TAMRA, or HEX) (Invitrogen) and 1U enzyme (Takara). PCR amplification was performed using the following program: pre-denaturation at 95°C for 5 min, followed by 35 cycles of denaturation at 94°C for 10 s in each cycle , anneal for 30 s (the annealing temperature of primers A and R is 51°C, and the annealing temperature of primer Y is 45°C) and extend at 72°C for 30 s, and finally extend at 72°C for 8 min.

[0037] (3) The amplified prod...

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 primer for identifying Rhododendron xing'an and Rhododendron japonica, which comprises: primers A, R and / or Y. The identification methods include: 1) extraction of whole genomic DNA; 2) amplification using the above primer system using genomic DNA as a template; 3) capillary electrophoresis of the amplified product; 4) determination based on the length of the PCR product band. Using the above method alone to distinguish Rhododendron xing'an from Rhododendron japonica, the identification accuracy rate can reach 100%, and considering the results of primer R and primer Y comprehensively, the identification accuracy rate can also reach 100%.

Description

technical field [0001] The invention belongs to the technical field of molecular biology molecular markers, and in particular relates to a primer and a method for identifying Rhododendron xing'an and Rhododendron japonica. Background technique [0002] Rhododendron ( Rhododendron L. ) plant ornamental value is very high, and primula, gentian and known as the world's three major alpine wild flowers. This genus is the largest in the Rhododendronaceae family and one of the largest in the Chinese and Himalayan flora. This genus occupies an important position in horticulture, and there are no less than 600 species of rhododendrons that have been introduced and cultivated in many countries in the world. Among them, the plants of the subgenus Rhododendron are small to medium-sized shrubs, deciduous or evergreen or semi-evergreen. This subgenus is generally considered to have two species: Rhododendron xing'an ( Rhododendron dauricum L.) and Rhododendron ( Rhododendron mucr...

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): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 肖洪兴王花英杨柏明孙明洲
Owner NORTHEAST NORMAL 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