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SNP (Single Nucleotide Polymorphism) molecular marker KQ8-3918 linked with gene for controlling Vc content of pepper fruits, application and special primer

A technology of molecular markers and gene linkage, applied in the direction of recombinant DNA technology, microbial measurement/testing, biochemical equipment and methods, etc., can solve the problems of accelerating the breeding of pepper varieties with high fruit vitamin C content and slow progress in molecular level research , to achieve the effect of improving breeding efficiency

Active Publication Date: 2022-01-04
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the metabolic pathways of capsicum involved in the biosynthesis, circulation, and degradation of fruit Vc have not yet been clarified, and the research on capsicum fruit Vc is limited to genetic rules and other aspects, and the progress of molecular level research is slow.
So far, there are no molecular markers that can be used to breed pepper varieties with high fruit vitamin C content. Therefore, the development of molecular markers that are closely linked to the genes that control the vitamin C content of pepper peppers, and the establishment of a molecular marker-assisted breeding technology system will help accelerate the development of peppers with high fruit vitamin C content. Breeding of varieties is very important

Method used

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  • SNP (Single Nucleotide Polymorphism) molecular marker KQ8-3918 linked with gene for controlling Vc content of pepper fruits, application and special primer
  • SNP (Single Nucleotide Polymorphism) molecular marker KQ8-3918 linked with gene for controlling Vc content of pepper fruits, application and special primer
  • SNP (Single Nucleotide Polymorphism) molecular marker KQ8-3918 linked with gene for controlling Vc content of pepper fruits, application and special primer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Acquisition of the Molecular Marker KQ8-3918 Linked to the Gene Linked to the Controlling Capsicum Fruit Vc Content

[0053] 1. Group construction

[0054] The F10 recombinant inbred lines containing 252 lines constructed by using inbred line Z5 and inbred line Z6 as parents were used as test materials. The fruit Vc content of semi-wild germplasm Z5 was 390mg / 100g fresh weight, and the fruit Vc content of sweet pepper inbred line Z6 was 25mg / 100g fresh weight.

[0055] 2. Vc content measurement:

[0056] When the capsicum fruit enters the red maturity stage, the fruit of each strain in the red maturity stage is mixed and sampled, and each strain has at least 15g of capsicum fruit, and the Vc content of the capsicum fruit in each strain is detected by a chromatograph.

[0057] 3. DNA extraction and SLAF data analysis:

[0058] Genomic DNA of each strain of parental and recombinant inbred lines populations was extracted using the Tiangen Plant Genomic DNA Ext...

Embodiment 2

[0079] Example 2 Reliability verification of molecular markers linked to genes controlling fruit Vc content in peppers

[0080] For the molecular markers developed in Example 1, further population verification was carried out. Taking 100 capsicum inbred lines bred by the inventor's research group as verification materials, the specific primers of Example 1 were used to carry out PCR amplification, and the molecular marker KQ8-3918 closely linked to the gene controlling the Vc content of the fruit was applied to the molecule The accuracy of marker-assisted breeding was verified.

[0081] The PCR reaction was carried out on a Hydrocycler water bath PCR instrument, and the fluorescence detection was carried out on the BMG LABTECH GMbH platform.

[0082] The PCR amplification system is: Genomic DNA (20ng / μL) 2.5ul, 2xKASP Master mix (purchased from Gene Company) 2.5μL; : 2:5 mixed first forward primer, second forward primer and universal reverse primer) 0.07 μL;

[0083] The am...

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Abstract

The invention belongs to the technical field of pepper variety breeding, and relates to an SNP (Single Nucleotide Polymorphism) molecular marker KQ8-3918 closely linked with a gene for controlling the Vc content of pepper fruits, a special primer for amplifying the molecular marker, a kit and application of the molecular marker KQ8-3918 and the special primer and the kit. A pepper high-density complete linkage genetic map is constructed for an RILs group containing 252 strains through an SLAF-seq technology, phenotypic data of the Vc content of fruits of all the strains of RILs in three seasons are combined, major QTL for controlling the Vc content of the fruits is positioned to a chromosome 1, a closely-linked SNP molecular marker KQ8-3918 is obtained, and the marker is located at the chromosome 1 32485915bp. A specific amplification primer developed by using the molecular marker can efficiently and accurately identify whether a pepper material or variety belongs to a material or variety with high fruit Vc content, and the breeding efficiency can be remarkably improved.

Description

technical field [0001] The invention relates to the technical field of capsicum variety breeding, in particular to a SNP molecular marker KQ8-3918 linked to a gene controlling the Vc content of capsicum fruit, special primers for amplifying the molecular marker, a kit and their application. Background technique [0002] Capsicum is an important vegetable and economic crop widely cultivated in the world, and it is also the main economic pillar crop in many areas of our country. With the continuous improvement of people's living standards, consumers' demand for high-quality agricultural products is more urgent, and the quality of vegetables is receiving more and more attention. The quality improvement of pepper has become an important research topic in pepper breeding. [0003] Pepper is recognized as the vegetable with the highest fruit vitamin C (Vc) content, and the fruit Vc content is an important nutritional quality of pepper. However, there were significant differences...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2600/13C12Q2531/113C12Q2535/125C12Q2563/107
Inventor 张晓芬耿三省陈斌杜和山王怡心
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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