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SNP locus closely related to orientation of capsicum fruit and its universal molecular marker, acquisition method and application thereof

A molecular marker and general-purpose technology, applied in the field of pepper genetics and breeding and molecular biology, can solve the problem of few molecular genetic mechanisms

Active Publication Date: 2019-10-08
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In pepper genetics and breeding research, fruit orientation is often used as a morphological marker, but there are relatively few studies on the molecular genetic mechanism of this trait variation
There have been no reports on the use of large-scale genome resequencing of pepper germplasm, nor the development of molecular markers closely associated with pepper orientation using genome-wide association analysis and linkage mapping of segregating populations

Method used

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  • SNP locus closely related to orientation of capsicum fruit and its universal molecular marker, acquisition method and application thereof
  • SNP locus closely related to orientation of capsicum fruit and its universal molecular marker, acquisition method and application thereof
  • SNP locus closely related to orientation of capsicum fruit and its universal molecular marker, acquisition method and application thereof

Examples

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Embodiment 1

[0049] This embodiment provides a SNP site closely associated with the orientation of pepper fruit, and the SNP site corresponds to base 37458043 of chromosome 12 of version 2.0 of the "Zunla 1" reference genome. Can be converted to CAPS markers.

[0050] This embodiment also provides a universal molecular marker CaUP12 closely related to the orientation of capsicum fruit, said molecular marker is transformed from genotyping primers designed according to the above SNP loci.

[0051] The primers mentioned above include forward primer CaUP12-F and reverse primer CaUP12-R, and the forward and reverse primers are as follows:

[0052] Forward primer CaUP12-F: 5'-CCAAGTCCCTAGATGGTGGTG-3', (SEQ ID NO.1);

[0053] Reverse primer CaUP12-R: 5'-TGCACAAGGACGTAGGTGTC-3', (SEQ ID NO. 2).

[0054] The above-mentioned universal molecular marker CaUP12 closely associated with the orientation of capsicum fruit is obtained by the following method, which includes the following steps:

[0055] ...

Embodiment 2

[0067] This example provides the application of the capsicum fruit oriented molecular marker CaUP12 in the above-mentioned Example 1 in natural populations.

[0068] In this example, the obtained fruit orientation molecular marker primers were used to identify the genotypes of 113 GWAS pepper materials, and predict the fruit orientation phenotype through the identification results. The steps are as follows:

[0069] (1) Genomic DNA of pepper was used as a template at a concentration of 80-150ng / μl; molecular marker CaUP12 was used for PCR amplification, the forward primer sequence was SEQ ID NO.1, and the reverse primer sequence was SEQ ID NO.2. The total volume of the PCR reaction system is 20 μl, and the specific components are as follows: ddH 2 O 15.6μl, 10×PCR Buffer 2.0μl, d NTPs (10mM) 0.4μl, TaqDNA polymerase 0.2μl (5U / μl), forward primer (10mM) 0.4μl, reverse primer (10 10mM) 0.4μl and 1.0 μl DNA template;

[0070] (2) The PCR reaction was carried out on the S1000 PC...

Embodiment 3

[0081] This example provides the application of the capsicum fruit oriented molecular marker CaUP12 in the above-mentioned Example 1 in the isolation population.

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Abstract

The present invention provides a mononucleotide polymorphism (SNP) loci highly associated with fruit orientation obtained by using genomic resequencing data and genome-wide association analysis (GWAS). A F2 segregating population with fruit orientation traits is used, by constructing two extreme mixing pools facing the phenotype, RNA sequencing (BSR) is carried out on the mixed pools, the SNP locus closely linked to the orientation of the capsicum fruit is obtained, the molecular markers obtained by the two strategies are used to finely position the orientation traits, and a tightly linked enzyme-cleaved polymorphic sequence (CAPS) marker CaUP12 is developed. CaUP12 is used to identify capsicum germplasm resources, and the accuracy rate reached 95.6%. The F2 segregating population is identified by CaUP12, and the accuracy rate reached 100%. The present invention provides the closely related molecular marker for the cloning of capsicum fruit control genes and capsicum breeding, which has important application value.

Description

technical field [0001] The invention belongs to the fields of capsicum genetics, breeding and molecular biology, and in particular relates to a SNP site closely related to capsicum fruit orientation and its general molecular marker, acquisition method and application. Background technique [0002] Capsicum (Capsicum annuum L.), also known as capsicum, capsicum annuum L., etc., belongs to the Solanaceae capsicum genus annual or perennial plant, is a class of important vegetable crops, widely planted around the world. Research evidence shows that pepper originated in South America and was introduced to my country in the late Ming Dynasty (16th century). As a vegetable, pepper is rich in nutrients such as vitamin C, E, minerals and carotenoids, and has high edible value. At the same time, pepper fruit contains unique capsaicinoids, which have anti-inflammatory and antioxidant effects. Because pepper has a spicy taste, it can promote the secretion of adrenaline, improve metabol...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 欧阳波陈蓉于会洋李峰陈文超欧立军邹学校
Owner HUAZHONG AGRI UNIV
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