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CN-122012771-A - Molecular marker and primer for identifying pulp color of dragon fruit and application of molecular marker and primer

CN122012771ACN 122012771 ACN122012771 ACN 122012771ACN-122012771-A

Abstract

The invention discloses a molecular marker and a primer for identifying the pulp color of dragon fruits and application thereof, and relates to the technical field of biology. The nucleotide sequence of the SNP molecular marker is shown as SEQ ID NO. 1. The molecular marker can be used for accurately identifying the pulp color of the dragon fruit. Meanwhile, the genetic improvement progress of the dragon fruit variety can be accelerated by applying the strain to molecular marker assisted selection (MARKER ASSISTED selection, MAS) and genome selection (Genomic selection, GS). The invention adopts KASP detection technology, the detection result is accurate and reliable, the detection method is simple to operate, the cost is low, the environment is friendly, no harm is generated to human body, and the invention is suitable for high-flux and large-scale commercial application.

Inventors

  • WU ZHIJIANG
  • WEI SHUOTONG
  • Liu chaoan
  • LI ZHENYING
  • HUANG ZEJIAN
  • LIANG GUIDONG
  • LU GUIFENG
  • DENG HAIYAN
  • LI HONGLI
  • HU WENBIN
  • HUANG LIFANG
  • YE XIAOYING

Assignees

  • 广西壮族自治区农业科学院
  • 中国热带农业科学院热带作物品种资源研究所

Dates

Publication Date
20260512
Application Date
20260127

Claims (10)

  1. 1. The SNP molecular marker related to the pulp color property of the dragon fruit is characterized in that the nucleotide sequence of the SNP molecular marker is shown as SEQ ID NO.1, and the SNP is positioned at the 150 th C > G single nucleotide variation from the 5' end of the SEQ ID NO. 1.
  2. 2. The primer group for detecting the SNP molecular marker according to claim 1, wherein the nucleotide sequence of the primer group is shown as SEQ ID NO. 2-4.
  3. 3. A kit for detecting the SNP molecular marker of claim 1, the kit comprising the primer set of claim 2.
  4. 4. The kit of claim 3, further comprising enzymes and dNTPs for use in a PCR reaction.
  5. 5. A genetic breeding method for screening the color characters of pitaya pulp is characterized by comprising the following steps of detecting genotypes of SNP molecular markers according to claim 1, and selecting individuals with different genotypes as parents according to breeding targets.
  6. 6. The method according to claim 5, wherein the method for detecting the genotype of the SNP molecular marker according to claim 1 comprises the steps of (1) extracting genomic DNA of a population of pitaya resources to be detected, (2) detecting the genotype of the pitaya to be detected by using the primer set according to claim 2 or the kit according to claim 3, and (3) determining the genotype of the molecular marker according to claim 1 of the pitaya to be detected based on the detection result.
  7. 7. A method for predicting the color property of pitaya pulp by using a molecular marker is characterized by comprising the following steps of detecting the genotype of the SNP molecular marker according to claim 1 and predicting the color of the pitaya pulp according to the detected genotype result.
  8. 8. The method according to claim 7, wherein the method for detecting the genotype of the SNP molecular marker according to claim 1 comprises the steps of (1) extracting genomic DNA of a sample of a dragon fruit to be detected, (2) detecting the genotype of the dragon fruit to be detected by using the primer set according to claim 2 or the kit according to claim 3, and (3) determining the genotype of the molecular marker according to claim 1 of the dragon fruit to be detected based on the detection result.
  9. 9. The use of the SNP molecular marker of claim 1 in dragon fruit pulp color trait related breeding or predicting dragon fruit pulp color.
  10. 10. Use of the primer set of claim 2 or the kit of claim 3 in breeding or predicting the color of the pulp of a dragon fruit in relation to the color trait of the pulp of a dragon fruit.

Description

Molecular marker and primer for identifying pulp color of dragon fruit and application of molecular marker and primer Technical Field The invention belongs to the technical field of biology, and particularly relates to a molecular marker and a primer for identifying the pulp color of dragon fruits and application thereof. Background The dragon fruit is taken as the tropical and subtropical special fruit, is favored by rich nutrition and unique taste, and the pulp color is one of the core characters for determining the commodity value, so that the market positioning and consumption demand difference of varieties with different colors are obvious. Almost no accumulation of colored secondary metabolites exists in the white-series pulp (the pulp color is white), but the red-series pulp (the pulp color is red or pink) is mainly caused by the accumulation difference of colored secondary metabolites (such as betaine and the like), and double-colored pulp also appears in rare cases (the double-colored pulp is red at the edge where the pulp contacts with the pericarp, and the inner part is mostly white, so the white-series pulp can also be used as the white-series pulp). However, the traditional breeding and pulp color judgment of the dragon fruits have a plurality of limitations that pulp color can be observed after plant results are obtained, the seedling stage can not be accurately judged through phenotypes, invalid plants need to be cultivated for a long time in the breeding process, resources such as land, manpower and the like are wasted, the traditional breeding relies on phenotype selection, the traditional breeding is easily interfered by environmental factors, the breeding period is as long as 3-5 years, and the efficiency is low. The molecular marking technology provides a new path for improving the crop properties by virtue of the advantages of accuracy and early detection. Through the molecular marker closely linked with the target trait, individuals containing the target trait can be rapidly screened in the seedling stage, the breeding period is shortened, and the selection efficiency is improved. At present, the research on specific molecular markers of the pulp color of the dragon fruit is less, and the lack of the accurate markers which can be directly applied to breeding practice restricts the development of directional breeding of the pulp color of the dragon fruit. Therefore, the molecular markers closely related to the color traits of the pulp of the dragon fruits are developed, a high-efficiency detection system is constructed, the accurate prediction and directional breeding of the pulp color in the seedling stage are realized, and the method has important significance for promoting the high-quality development of the dragon fruit industry. Disclosure of Invention Aiming at the problems of judgment after the result is needed for the pulp color of the dragon fruit, long breeding period, low efficiency and the like in the prior art, the invention develops SNP molecular markers which can be used for identifying the pulp color of the dragon fruit through a high-throughput sequencing technology, and verifies the SNP molecular markers in genetic groups and natural groups, the markers can be used for commercial identification of the dragon fruit (seedlings) sold in the market, and simultaneously, the genetic improvement progress of the dragon fruit variety can be accelerated by applying the markers to molecular marker assisted selection (MARKER ASSISTED selection, MAS) and genome selection (Genomic selection, GS). The primary aim of the invention is to determine SNP molecular markers related to pulp color traits, wherein the molecular markers are positioned on a pitaya genome (http:// www.pitayagenomic.com/download. Php), the nucleotide sequence of the molecular markers is shown as SEQ ID NO.1, the SNP is positioned at a SNP mutation of 150 th position C > G from the 5' end of the SEQ ID NO.1, the mutation and the pulp color form have stable corresponding relation, namely, GG type is mainly white pulp, CG or CC type is mainly red or pink pulp, the CC genotype is preferred if a red pulp phenotype is required to be obtained, and the CG genotype is preferred if a pink pulp phenotype is required to be obtained. In order to realize efficient and accurate detection of the molecular marker, the invention provides a set of specific primer groups adapting to KASP (competitive allele-specific PCR) typing technology, which is characterized in that the nucleotide sequences of the primer groups are respectively shown as SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4, wherein the SEQ ID NO.2 and the SEQ ID NO.3 are allele-specific primers, the 5' ends of the primers are respectively connected with fluorescent labels with different colors, and the SEQ ID NO.4 is a universal primer. The primer set is designed for SEQ ID NO.1 sequence and 150 SNP loci, can specifically identify target loci through KASP typing technology wi