Unlock instant, AI-driven research and patent intelligence for your innovation.

KASP molecular marker for detecting rice blast resistance gene Pi25, and application thereof

A rice blast resistance gene and molecular marker technology, applied in the field of agricultural biology, can solve the problems of many specific markers, low polymorphism rate of linked markers, unstable amplification, etc., to eliminate aerosol pollution, save time and Labor cost, environment-friendly effect

Inactive Publication Date: 2021-02-26
SHANGHAI AGROBIOLOGICAL GENE CENT
View PDF6 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In practical applications, CAPS markers require enzyme digestion, and the process is cumbersome; linkage markers have the disadvantages of low polymorphism rate and small differences, more restrictive conditions for specific markers, and unstable amplification. The above markers used in the detection process EB or polyacrylamide is easy to pollute the environment and cause harm to the human body

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
  • KASP molecular marker for detecting rice blast resistance gene Pi25, and application thereof
  • KASP molecular marker for detecting rice blast resistance gene Pi25, and application thereof
  • KASP molecular marker for detecting rice blast resistance gene Pi25, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Whole-genome DNA was extracted from Gumei 2 and Hui 12-33, other blast-resistant donor materials, and 38 water-saving and drought-resistant rice parents (45 parents in total) without the Pi25 gene by CTAB method;

[0061] The KASP reaction was performed on the whole genome DNA of the above parents (the following 1) and 2) do not have a sequence relationship, and there is no order limitation):

[0062] 1) Use the Pi25-K775 primer set to perform PCR amplification on the ABI 7500 real-time PCR system using the whole genome DNA of the above 45 parents as a template.

[0063] The total volume of the PCR reaction system was 10 μL, including 5 μL of 2×KASP Master mix and 0.14 μL of KASP Primer mix (the ratio of Pi25-K775-FAM, Pi25-K775-HEX and Pi25-K775-Common primer was 1:1:2.5 ), template DNA 1μL (20ng / μL), ddH 2 O 3.86 μL.

[0064]The PCR reaction program is 94°C pre-denaturation for 15 minutes; 94°C denaturation for 20s; 65°C-57°C annealing and extension for 60s, 10 cycl...

Embodiment 2

[0077] Application of KASP Molecular Marker of Rice Blast Resistance Gene Pi25

[0078] 1. Genotype identification of natural populations

[0079] In view of the advantages of the molecular marker design described in the present invention, the KASP reaction can be used for high-throughput identification of the rice blast resistance Pi25 genotype of rice germplasm resources, and the two pairs of KASP molecular markers described in the present invention were used to identify the 270 rice Genotype analysis was performed on the DNA of the micro-core germplasm population (Wu et al.BMC plant biology, 2015,15(1):218), and it was found that there were 4 local varieties with Pi25 resistance pure alleles (HONGWAN 1, SHUIYUAN300LI, SANBAILI, NANXIONGZAOYOU), these 4 materials can be used as Pi25 gene donor parents in marker-assisted breeding.

[0080] 2. Genotype identification of genetic segregation populations

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 provides a KASP molecular marker for detecting a rice blast resistance gene Pi25, and application thereof, and belongs to the technical field of agricultural biology. According to the invention, two pairs of primers Pi25-K775 and Pi25-K2680 / 2687 are designed according to key specific sites of a target gene at 775bp and 2687bp of a Pi25 gene coding region; PCR amplification is carriedout by taking rice genome DNA as a template; then signal scanning analysis is carried out on an amplification product; and genotyping on the target gene is carried out. According to the invention, The KASP markers designed on the basis of the KASP reaction principle and the single base difference in the anti-infection material Pi25 gene coding region can be used for performing Pi25 gene detectionon a rice material in a high-throughput mode; the two pairs of KASP molecular markers can be verified mutually; errors in the experiment process are avoided; higher accuracy is achieved; and the KASPmolecular marker has the remarkable advantages of simplicity, convenience, reliability, quickness, strong specificity, high sensitivity and environmental friendliness, and is suitable for identification and typing of the Pi25 gene and auxiliary selective breeding of rice blast resistant varieties.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and in particular relates to a KASP molecular marker for detecting rice blast resistance gene Pi25 and its application. Background technique [0002] Rice blast is one of the most devastating fungal diseases in rice production at present. It is distributed and occurs continuously in all rice-growing areas in the world, seriously affecting rice production and bringing great hidden dangers to food security. Practice has shown that cultivating and promoting rice blast-resistant varieties is the most economical, effective, safe and environmentally friendly measure to control rice blast. [0003] Traditional rice disease resistance breeding is phenotypic selection of plants through resistance identification, which takes a long time and is easily limited by environmental conditions. The identification results are prone to errors and the selection efficiency is low. The use of molecular marker...

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): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2531/113C12Q2563/107
Inventor 刘毅王飞名张安宁孔德艳刘国兰余新桥罗利军
Owner SHANGHAI AGROBIOLOGICAL GENE CENT