A SNP site closely associated with pepper fruit orientation and its general molecular marker, acquisition method and application

A technology of molecular markers and application methods, applied in the fields of pepper genetics, breeding and molecular biology, can solve the problems of few molecular genetic mechanisms and the like

Active Publication Date: 2021-02-12
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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  • A SNP site closely associated with pepper fruit orientation and its general molecular marker, acquisition method and application
  • A SNP site closely associated with pepper fruit orientation and its general molecular marker, acquisition method and application
  • A SNP site closely associated with pepper fruit orientation and its general molecular marker, acquisition method and application

Examples

Experimental program
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Effect test

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 uses genome resequencing data and genome-wide association analysis (GWAS) to obtain single nucleotide polymorphism (SNP) sites that are highly correlated with fruit orientation, and at the same time utilizes the F2 segregation population of fruit orientation traits to construct orientation phenotypes The two extreme mixed pools were combined with RNA sequencing (BSR) on the mixed pools to obtain the SNP loci closely linked to the orientation traits of pepper fruit, and the molecular markers obtained by the two strategies were used to fine-tune the orientation traits, and one was developed. Tightly linked cleavage amplified polymorphic sequence (CAPS) marker CaUP12. Using CaUP12 to identify pepper germplasm resources, the accuracy rate reached 95.6%, and using CaUP12 to identify F2 segregation populations, the accuracy rate reached 100%. Therefore, the present invention provides closely related molecular markers for the cloning of pepper fruit orientation control genes and pepper breeding, and 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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Patent Type & Authority Patents(China)
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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