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Downy mildew resistance gene BrRLP48 on Chinese cabbage A04 chromosome and SNP (single-nucleotide polymorphism) marker linked to same

A technology for cabbage downy mildew and downy mildew resistance, applied in DNA/RNA fragments, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problem of long cycle of disease-resistant cabbage

Active Publication Date: 2019-01-04
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Genetic studies have shown that cabbage downy mildew resistance is a complex quantitative trait that involves multiple disease resistance genes, and it takes a long time to obtain disease-resistant cabbage by relying on traditional breeding methods
With the development of molecular biology, molecular assisted breeding technology can carry out accurate and stable selection in early generations, thereby speeding up the breeding process and improving breeding efficiency, but the basis of this technology is to have excellent target genes and closely linked molecules mark

Method used

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  • Downy mildew resistance gene BrRLP48 on Chinese cabbage A04 chromosome and SNP (single-nucleotide polymorphism) marker linked to same
  • Downy mildew resistance gene BrRLP48 on Chinese cabbage A04 chromosome and SNP (single-nucleotide polymorphism) marker linked to same
  • Downy mildew resistance gene BrRLP48 on Chinese cabbage A04 chromosome and SNP (single-nucleotide polymorphism) marker linked to same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] Example 1, the acquisition of SNP marker A04_5326733

[0104] 1. Test materials

[0105]F 1 The microspores of the population were obtained for chromosome doubling, and finally a doubled haploid population (DH, Doubled Haploid population) was obtained, and the DH population contained 80 individual plants.

[0106] F 1 Generation groups, the F 1 Each individual plant in the generation population is self-crossed, and F 2 group, F 2 The population contained 1056 individuals.

[0107] The DH population consisting of 80 strains and the F population consisting of 1056 strains were selected. 2 groups as targeting groups.

[0108] 2. Genotyping

[0109] Planting two parental materials in the greenhouse, the disease-resistant parent "MM" and the susceptible parent "Baiyang", F 1 generation group, F 2 Population and DH population plant material. The young leaves were taken, and the whole genome DNA was extracted according to Doyle and Dickson (1987) with a slightly mod...

Embodiment 2

[0126] Example 2, Application of A04_5326733 locus in identification of cabbage downy mildew resistance

[0127] 1. Test materials

[0128] With the F that is made up of 1056 strains in embodiment 1 2 The population, the disease-resistant inbred line "MM", and the susceptible inbred line "Baiyang" were used as test materials.

[0129] 2. Identification of downy mildew resistance

[0130] The downy mildew resistance of the tested materials was identified. The specific steps are as follows: plant the test material in the greenhouse, and inoculate downy mildew on the 16th day of planting when most of the two true leaves of the plants are fully unfolded and new true leaves are exposed (that is, the two-leaf one-core stage). Each mL will contain 1 x 10 4 or 1×10 5 The suspension of downy mildew pathogenic spores is evenly sprayed on the back of the leaves by spraying. Place the inoculated plants in a dark, 100% humidity environment at 18-20°C for 24 hours. Subsequently, move...

Embodiment 3

[0165] Example 3, disease resistance gene BrRLP48 and its functional analysis

[0166] 1. Transient expression experiment of disease resistance gene BrRLP48

[0167] There are 16 functionally annotated genes in the 160kb Bra-DM04 interval. According to the annotations, there are four possible candidate genes for disease resistance. The proteins encoded by these candidate genes all contain Leucine-rich repeat (LRR, Leucine-rich repeat) domains , because the marker A04_5326733 is inside the disease resistance candidate gene BrRLP48, which encodes a transmembrane protein containing 7 LRR (leucine-rich repeat) domains, and belongs to the receptor-like protein (RLP, receptor-like protein) family. Therefore, the BrRLP48 candidate gene was applied to the transient expression experiment of cotyledons of Chinese cabbage to verify its disease resistance function. The BrRLP48 candidate gene sequence is shown in sequence 6, the CDS sequence is shown in sequence 7, and the amino acid sequ...

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Abstract

The invention discloses downy mildew resistance gene BrRLP48 on Chinese cabbage A04 chromosome and an SNP (single-nucleotide polymorphism) marker linked to the same. DH and F2 populations of Chinese cabbage disease-resistant inbred line MM and susceptible inbred line Bai Yang are used herein to successfully locate QTL (quantitative trait locus) Br-DM04 related to Chinese cabbage downy mildew resistance on Chinese cabbage No. 4 chromosome; 22.3% of phenotypic variations can be interpreted, the downy mildew resistance gene and the SNP molecular marker A04_5326733 linked thereto can be located. The molecular marker A04_5326733 based on KASP (competitive allele-specific PCR) technique is applicable to identification and molecular breeding of Chinese cabbage downy mildew resistant breeds.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the downy mildew resistance gene BrRLP48 located on the chromosome of Chinese cabbage A04 and the SNP marker linked to it. Background technique [0002] Chinese cabbage is native to my country and is the vegetable crop with the largest cultivated area in my country. It has gradually become a worldwide vegetable crop. According to the statistics of the Ministry of Agriculture, the annual planting area of ​​Chinese cabbage in my country is nearly 40 million mu, accounting for about 15% of the total planting area of ​​vegetables in the country. It can be seen that Chinese cabbage plays a decisive role in my country's vegetable basket project. Plant diseases are one of the major threats to the normal growth of plants. Especially in agricultural production, diseases will directly affect the yield and quality of crops. Downy mildew is a fungal disease caused by obligate paras...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 于拴仓张彬苏同兵张凤兰李佩荣李盼余阳俊张德双赵岫云汪维红
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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