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A molecular marker closely linked to rice blast resistance gene, primer and application thereof

A rice blast and molecular marker technology, applied in the field of molecular markers and primers, can solve the problems of reduced yield and smaller rice seeds, and achieve the effects of reducing labor land, shortening breeding cycle, and improving breeding efficiency.

Inactive Publication Date: 2020-05-26
安徽丰大种业股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

PigmR has broad-spectrum disease resistance to all detected Magnaporthe grisea races, but at the same time makes rice seeds smaller and lowers yield; PigmS inhibits the disease resistance function of PigmR, but can increase rice yield

Method used

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  • A molecular marker closely linked to rice blast resistance gene, primer and application thereof
  • A molecular marker closely linked to rice blast resistance gene, primer and application thereof
  • A molecular marker closely linked to rice blast resistance gene, primer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Obtaining the nucleotide sequence shown in SEQ ID NO: 3 and the nucleotide sequence shown in SEQ ID NO: 4 by the special primer pair for identification of rice blast resistance traits

[0026] 1.1 Parental Genome Amplification

[0027] The parents used to create the rice recombinant inbred line (recombinant inbred line, RIL) population were Gumei 4 and FD1715 bred by Anhui Fengda Seed Industry Co., Ltd., respectively. FD1715 is the female parent, which is not resistant to rice blast; Gumei 4 is the male parent, which is highly resistant to rice blast.

[0028] Genomic DNA of parental leaves was extracted by CTAB method, and primers were designed according to the sequence published by Yiwen Deng (Yiwen Deng, et al.Epigenetic regulation of antagonistic receptors confers rice blast resistance with yield balance[J].2017.355(6328):962). The nucleotide sequence shown in SEQ ID NO: 5 and the nucleotide sequence shown in SEQ ID NO: 6 are composed of the nucleotide sequence sho...

Embodiment 2

[0036] Correlation verification between the amplified product of QK1 and rice blast resistance traits

[0037] 432 F 2 The substitute material is the test object. Genomic DNA was extracted from the rice to be tested by the alkaline cooking method, and the specific steps were the same as in Example 1. Perform PCR amplification experiments. The PCR reaction system is: 10X Buffer 2μl (containing Mg 2+ ), dNTP 0.4μl (10mM), the nucleotide sequence shown in SEQ ID NO: 3 and the nucleotide sequence shown in SEQ ID NO: 4 each 2μl (5μM), genomic DNA extracted from the rice to be tested 1μl , taq enzyme 0.5μl (5U / μl), the rest is ultrapure water, the reaction volume is 20μl, and a drop of mineral oil is added dropwise to cover. The reaction program of PCR amplification is: 94°C for 5 minutes; 94°C for 60s, 53.5°C for 60s, 72°C for 60s, 35 cycles; 72°C for 10min.

[0038] PCR amplified products were analyzed by 6% polyacrylamide gel electrophoresis, stained by rapid silver staining...

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Abstract

The invention discloses a molecular marker tightly linked with a paddy rice blast resistance gene, a primer pair amplifying the molecular marker and a method of detecting paddy rice blast resistance. A nucleotide sequence of the molecular marker is shown in SEQ ID NO: 1 or SEQ ID NO: 2. The molecular marker is located in a resistance gene, a Pigm gene, and tightly linked with the paddy rice blast resistance gene; compared with a non-genomic internal marker, the molecular marker can not be separated, so that the seed breeding efficiency can be improved more effectively.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a molecular marker closely linked with a rice blast resistance gene, a primer and an application thereof. Background technique [0002] Conventional breeding methods based on phenotypic selection have disadvantages such as low selection efficiency and long breeding cycle. It is urgent to inject modern molecular technology, supplemented by high-efficiency genotype-directed selection, in order to quickly and efficiently breed excellent new rice varieties. With the rapid development of molecular biology and genomics, the application of molecular marker technology is more extensive. PCR-based molecular markers such as microsatellites or SSR (simple sequence repeat) are widely used because of their high polymorphism rate, relative stability, simple and fast detection method, and easy operation. [0003] Since molecular marker-assisted selection is not easily affected by environmental fac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 何立斌朱先飞李婧婧张志清付求来潘华周健李有强
Owner 安徽丰大种业股份有限公司
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