CN-121406823-B - Indel molecular marker closely related to content of soluble sugar in capsicum fruits and application of Indel molecular marker

CN121406823BCN 121406823 BCN121406823 BCN 121406823BCN-121406823-B

Abstract

The invention provides an Indel molecular marker closely related to the content of soluble sugar in pepper fruits and application thereof, belonging to the technical field of determination methods containing nucleic acid. The nucleotide sequence of the Indel molecular marker is shown as SEQ ID NO.1 in a sequence table, and the Indel molecular marker is positioned at 5274716-5274727 th site of the full-length sequence of chromosome 3 of the capsicum reference genome. The primer for amplifying the Indel molecular marker comprises a forward primer and a reverse primer, wherein the nucleotide sequence of the forward primer is shown as SEQ ID NO.2 in a sequence table, and the nucleotide sequence of the reverse primer is shown as SEQ ID NO.3 in the sequence table. The molecular marker can be directly applied to early generation of breeding, so that destructive sampling of chemical detection is avoided, the breeding cost is obviously reduced, the breeding period is effectively shortened, and the selection efficiency of auxiliary breeding of the capsicum molecular marker is greatly improved.

Inventors

GAO JUEXIAO
YUAN YANFANG
LI HUIRU
CAO HUI
LIANG ZENGWEN
DING ZHUO
LIANG XIYUAN
JIANG QIANQIAN
XIN GUOBIN
GAO MINGGANG
LI JINLING
QI YULIANG

Assignees

山东永盛农业发展有限公司

Dates

Publication Date: 20260505
Application Date: 20251226

Claims (4)

1. The Indel molecular marker closely related to the content of the soluble sugar in the pepper fruits is characterized in that the nucleotide sequence of the Indel molecular marker is shown as SEQ ID NO.1 in a sequence table, the Indel molecular marker is positioned at 5274716-5274727 th site of the full-length sequence of chromosome 3 of a pepper reference genome, and the version of the pepper reference genome is ASM51225v2.
2. The Indel molecular marker closely related to the soluble sugar content of pepper fruits according to claim 1, wherein the primer for amplifying the Indel molecular marker comprises a forward primer and a reverse primer, the nucleotide sequence of the forward primer is shown as SEQ ID NO.2 in the sequence table, and the nucleotide sequence of the reverse primer is shown as SEQ ID NO.3 in the sequence table.
3. Use of the primer for amplifying the molecular marker of claim 2 for identifying the soluble sugar content of pepper fruits, comprising the steps of: (1) Extracting genome DNA of pepper germplasm leaves to be detected; (2) Taking genome DNA of pepper germplasm leaves as a template, carrying out PCR amplification by using primers of an amplification molecular marker, and detecting PCR amplification products; (3) If the molecular marker is amplified to obtain an amplified product with the length of 219bp, the content of the fruit soluble sugar of the pepper germplasm is predicted to be more than or equal to 4.5 percent, and if the molecular marker is not provided with an amplified band, the content of the fruit soluble sugar of the pepper germplasm is predicted to be less than 4.5 percent.
4. The method according to claim 3, wherein the nucleotide sequence of the 219bp amplification product is shown in SEQ ID NO.6 of the sequence Listing.

Description

Indel molecular marker closely related to content of soluble sugar in capsicum fruits and application of Indel molecular marker Technical Field The invention relates to an Indel molecular marker closely related to the content of soluble sugar in pepper fruits and application thereof, belonging to the technical field of determination methods containing nucleic acid. Background The capsicum is used as an important vegetable and condiment, has rich nutrition and has important industrial value. The content of the soluble sugar in the capsicum can directly influence the flavor quality and stress resistance of the capsicum. Through phenotypic selection or molecular marker assisted breeding, the sugar content can be directionally improved, the sweet and spicy balance is optimized, the fruit storability is improved, high-sugar high-quality varieties are cultivated, the diversified requirements of the market on the flavor type chilli are met, and the accurate development of the breeding technology is promoted. The traditional breeding method for breeding the high-solubility sugar chilli variety has the defects of long period, dependence on phenotype selection, easiness in environmental interference and the like. The molecular marker detection can accurately locate sugar metabolism related genes, realize early screening, shorten breeding period, improve selection efficiency, overcome the limitation of unsound association of phenotype and genotype, and promote accurate breeding development. Indel (insertion-deletion) markers have the advantages of high polymorphism and whole genome coverage in the detection of pepper soluble sugar, can rapidly genotype through PCR-electrophoresis, accurately locate sugar metabolism related genes, have stable inheritance and simple detection, can early screen germplasm with high soluble sugar content, shorten breeding period, and improve selection efficiency, and are efficient and economic tools in molecular marker assisted breeding. There is no report on Indel molecular markers closely related to the soluble sugar content of pepper fruits in the prior art. Disclosure of Invention Aiming at the defects existing in the prior art, the invention provides the Indel molecular marker closely related to the content of the soluble sugar in the capsicum fruits and the application thereof, so that the early nondestructive screening of capsicum germplasm with high content of the soluble sugar is realized, the accuracy is high, and the breeding period is shortened. In order to solve the technical problems, the invention adopts the following technical scheme: An Indel molecular marker closely related to the content of soluble sugar in pepper fruits, wherein the nucleotide sequence of the Indel molecular marker is shown as SEQ ID NO.1 in a sequence table, and the Indel molecular marker is positioned at 5274716-5274727 th site of the full-length sequence of chromosome 3 of a pepper reference genome. The primer for amplifying the Indel molecular marker comprises a forward primer and a reverse primer, wherein the nucleotide sequence of the forward primer is shown as SEQ ID NO.2 in a sequence table, and the nucleotide sequence of the reverse primer is shown as SEQ ID NO.3 in the sequence table. The application of the primer for amplifying the molecular marker in the identification of the content of soluble sugar in pepper fruits. The application comprises the following steps: (1) Extracting genome DNA of pepper leaves to be detected; (2) Taking genome DNA of capsicum leaves as a template, carrying out PCR (polymerase chain reaction) amplification by using primers of an amplification molecular marker, and detecting PCR amplification products; (3) If the molecular marker is amplified to obtain an amplified product with the length of 219bp, predicting that the content of the fruit soluble sugar of the pepper germplasm is more than or equal to 4.5 percent, and is a pepper germplasm material with high content of soluble sugar, and if the molecular marker is not provided with an amplified band, predicting that the content of the fruit soluble sugar of the pepper germplasm is less than 4.5 percent, and is a pepper germplasm material with low content of soluble sugar. The nucleotide sequence of the 219bp amplification product is shown as SEQ ID NO.6 in the sequence table. Compared with the prior art, the invention has the following beneficial effects: The invention develops an Indel molecular Marker which is obviously related to the content of the soluble sugar in the pepper fruits, and can be used for auxiliary selection (Marker-Assisted Selection, MAS) breeding of the pepper molecular markers, so that early nondestructive screening is realized. In the early growth period of the capsicum, genotypes with high soluble sugar and low soluble sugar content can be rapidly distinguished by PCR amplification, so that capsicum varieties with high soluble sugar content can be bred efficiently and directionally. The mole