Apricot fruit firmness key gene molecular marker PG1524 and detection method and application thereof

A molecular marker, fruit technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of high average sample analysis price, high sample requirements, and cumbersome operation, and reduce planting. area, accelerated breeding process, low cost effect

Pending Publication Date: 2019-10-08
辽宁省果树科学研究所 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the high-throughput sequencing method has high detection efficiency, it has disadvantages such as expensive instruments, high average price of sample analysis, and high requirements for samples.
Although the cost of PCR-RFLP and PCR-SSCP is not high, the operation is cumbersome

Method used

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  • Apricot fruit firmness key gene molecular marker PG1524 and detection method and application thereof
  • Apricot fruit firmness key gene molecular marker PG1524 and detection method and application thereof
  • Apricot fruit firmness key gene molecular marker PG1524 and detection method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Acquisition of SNP molecular marker PG1524

[0028] (1) Download the PG gene sequence of apricot (Goldrich) (GenBank accession No.HG540310) and the PG gene sequence of peach (GenBankaccession No.) from GenBank of NCBI (website: http: / / www.ncbi.nlm.nih.gov) .DQ340809), using this as a template, use the software Primer premier 5.0 to design 2 pairs of gene-specific primers (PG-1 / PG-2, PG-3 / PG-4). The primer sequences are shown in Table 1.

[0029] Table 1 Primer sequence

[0030]

[0031] (2) According to the National Fruit Tree Germplasm, Xiongyue Li Xing Garden, apricot resources and fruit hardness survey data, two hard flesh varieties were selected: Sungold and Moopark; two soft flesh varieties were selected: Xianju apricot and Xiaosaimai, young and tender were collected in spring Leaves, using modified CTAB method to extract genomic DNA.

[0032] (3) Using the genomic DNA of the above four apricot varieties as templates, the primers (PG-1 / PG-2, PG-3 / PG-4) were use...

Embodiment 2

[0036] Example 2: Testing the fruit hardness traits of the genetic population "Camel Yellow x Golden Sun" at the seedling stage

[0037] (1) Construction of genetic population

[0038] A cross combination was configured with soft apricot variety ‘Luotuohuang’ as the female parent and hard apricot variety ‘Golden Sun’ as the male parent, and 25 F plants were obtained. 1 Generation hybrid population.

[0039] (2) The leaves of annual seedlings were collected in spring, and genomic DNA was extracted by modified CTAB method.

[0040] (3) Amplify with primer pair (F / R, P / Q) of molecular marker PG1524. The PCR reaction system is 20 μL, including 5.0 μL of DNA template, 10.0 μL of 2×Taq Master Mix (Beijing Kangwei Century Biological Company), 1.0 μL of positive and negative inner primers, 0.5 μL of positive and negative outer primers, and 2 μL of water. The PCR amplification procedure is as follows: 95°C for 5min; 94°C for 40s, 55°C for 40s, 72°C for 45s, 35 cycles; 72°C for 10min. The PCR...

Embodiment 3

[0043] Example 3: Testing the fruit hardness characteristics of 120 common apricot resources

[0044] (1) The leaves of 120 ordinary apricots were collected in spring, and the genomic DNA was extracted by the modified CTAB method.

[0045] (2) Amplify with primer pair (F / R, P / Q) of molecular marker PG1524. The PCR reaction system is 20 μL, including 5.0 μL of DNA template, 10.0 μL of 2×Taq Master Mix (Beijing Kangwei Century Biological Company), 1.0 μL of positive and negative inner primers, 0.5 μL of positive and negative outer primers, and 2 μL of water. The PCR amplification procedure is as follows: 95°C for 5min; 94°C for 40s, 55°C for 40s, 72°C for 45s, 35 cycles; 72°C for 10min. The PCR products were separated by electrophoresis on a 2% agarose gel, stained with ethidium bromide, and the gel imaging system scanned and saved the image.

[0046] (3) Count the genotype of each plant and predict its fruit hardness traits. The genotype containing fragment 1 (408bp) and fragment 2...

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Abstract

The invention discloses an apricot fruit firmness key gene molecular marker PG1524 and a detection method and application thereof. The molecular marker PG1524 is applicable to assisted selection for breeding of firm-pulp apricot varieties; identification on fruit firmness can be achieved in seedling stage; material screening can be performed on a large scale; phenotype measuring can be performed without waiting for apricot fruiting; identification time is advanced greatly; unnecessary planting area is narrowed; the consumption of manpower and material resources is reduced; the breeding progress of firm-pulp apricot varieties is quickened; the molecular marker PG1524 has important applicable value and social value. In addition, genotype detecting is performed through the bidirectional allele specific PCR (polymerase chain reaction) technique; SNP (single nucleotide polymorphism) typing of apricot plants can be achieved simply and quickly at low cost; utilization efficiency of the apricot fruit firmness key gene molecular marker PG1524 is improved.

Description

Technical field [0001] The invention belongs to the technical field of fruit tree molecular genetic breeding, and specifically relates to a molecular marker PG1524 of a key gene of apricot fruit hardness and a detection method and application thereof. Background technique [0002] Apricot (scientific name: Armeniaca vulgaris Lam.) is rich in various nutrients and is one of the important fruits in early summer. The hardness of most apricot fruits declines rapidly after ripening, leading to poor storage and transportation performance, short shelf life and perishable deterioration, which seriously affects the commodity value of apricot fruits and is a bottleneck restricting the development of apricot industry. In order to solve this problem, it is urgent to cultivate hard apricot varieties that are resistant to storage and transportation, so as to accelerate the development of the apricot industry. However, fruit trees are multi-year biological species with a long young age and a l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2600/172C12Q2531/113C12Q2565/125
Inventor 魏潇章秋平刘威生方林川
Owner 辽宁省果树科学研究所
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