Molecular marker for rapidly identifying density of spines of cucumber line, identification method and application

A molecular marker, cucumber technology, applied in the direction of biochemical equipment and methods, determination/inspection of microorganisms, DNA/RNA fragments, etc.

Active Publication Date: 2015-07-29
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Since there is no report on the positioning and cloning of the cucumber thorn tumor density gene, at present, the breeding work related to the cucumber thorn tumor density is still screened by the breeder's observation of the fruit thorn tumor traits after the cucumber bears fruit. Not only can personnel not select at the seedling stage, but it is also difficult to accurately transfer small fragments of tumor density genes

Method used

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  • Molecular marker for rapidly identifying density of spines of cucumber line, identification method and application
  • Molecular marker for rapidly identifying density of spines of cucumber line, identification method and application
  • Molecular marker for rapidly identifying density of spines of cucumber line, identification method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Example 1 Construction of genetic segregation population

[0024]The North China type cucumber inbred line 9930 was used as the female parent, and the North China type cucumber inbred line Shannong CNS5 was used as the male parent. The fruit phenotype of the female cucumber inbred line 9930 is dense thorn tumor; the male cucumber inbred line Shannong CNS5 is a natural mutant isolated from the North China type cucumber Nongcheng No. 2 inbred line Shannong CNS2. The phenotype of its fruit is sparse spiny tumor.

[0025] Using these two parents to prepare a cross combination, get F 1 The prickly tumor density of the fruit was between the two parents, F 1 F 2 78 strains were randomly selected for genetic segregation analysis.

[0026] In 78 F 2 Among individuals, the fruit prickly tumor density of each plant was counted, and the chi-square analysis was used to analyze the segregation ratio of dense prickly / sparse prickly traits. Finally, it was determined that the pric...

Embodiment 2

[0027] Example 2 Whole genome resequencing of cucumber

[0028] Using population segregation analysis (BSA) from F 2 In the isolated population, 20 individual plants of S. spp. and S. spp. were randomly selected, and the gene pools of S. spp. (fs1 fs1) ​​and S. spp. (FS1_) were established.

[0029] The constructed gene pools of cucumber with sparse tumor (fs1 fs1) ​​and cucumber with dense tumor (FS1_) were sent to Shanghai Meiji Biomedical Technology Co., Ltd. for whole genome resequencing of cucumber.

[0030] The association analysis between the SNP markers obtained by sequencing and the phenotypic traits of tumor density showed that the fs1 gene fs1, which controls the thin thorn phenotype of cucumber, was mapped to the end of the long arm of chromosome 6.

Embodiment 3

[0031] Example 3 Development of Molecular Markers

[0032] Analyze the SNP marker results obtained by resequencing the whole cucumber genome, design marker primers according to the position of the sparse thorn gene fs1, use the designed CAPS marker primers and SNP marker primers to perform PCR amplification on the two parents, and use 1.2% agarose Gel electrophoresis detection (the 1.2% agarose gel means that 100 ml of TBE buffer contains 1.2 g of agarose powder).

[0033] The following 6 markers were screened from the designed CAPS markers and SNP markers to carry out PCR amplification between the two parents. The results of the amplification primers, restriction enzymes and amplified fragments are shown in Table 1.

[0034] Table 1 Marker primer sequences and amplified fragment results of 6 kinds of molecular markers

[0035]

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Abstract

The invention discloses a molecular marker for rapidly identifying the density of spines of a cucumber line. The molecular marker is a molecular marker MM3, a molecular marker MM1 or a molecular marker MM2; the molecular marker MM3 consists of a nucleotide sequence shown in SEQ ID NO:9 and a nucleotide sequence shown in SEQ ID NO:10, wherein the nucleotide sequence shown in the SEQ ID NO:9 is totally linked with a cucumber sparse spine gene FS1; the nucleotide sequence shown in the SEQ ID NO:10 is totally linked with a cucumber sparse spine gene fs1; meanwhile, the invention also provides a method for rapidly identifying a cucumber variety or the density of the spines of the line by utilizing the molecular marker MM3. The molecular marker and the identification method provided by the invention are beneficial for building a molecular marking assisting breeding system for sparse/dense spine cucumbers and applying the molecular marking assisting breeding system to cucumber breeding practices and have the advantages of being rapid and accurate in screening, not affected by environment, definite in selected target, capable of lowering the production cost and greatly improving the selection efficiency and quality of a target cucumber variety or the spine type of the line, and quite high in social and economic values.

Description

technical field [0001] The invention relates to the technical field of biogenetic engineering, in particular to a molecular marker for rapidly identifying the density of spiny tumors of cucumber strains, an identification method and an application thereof. Background technique [0002] Cucumber is one of the horticultural crops with important economic value and is widely cultivated all over the world. Since the quality of cucumbers is directly related to the benefits of cucumber planting, high-quality breeding has always been the top priority in cucumber breeding. The types of cucumber varieties in my country are mainly divided into North China type and South China type. These two types of cucumbers have their own advantages and disadvantages. The main manifestations are that most of the South China type cucumbers are thin prickly tumor varieties, but the cucumber flavor is light; the North China type cucumber has a strong flavor, but Most of them are dense thorn tumor varie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/158
Inventor 任仲海张海洋曹辰兴王丽娜李强
Owner SHANDONG AGRICULTURAL UNIVERSITY
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