CAPS molecular marker closely linked with sepal color trait gene of solanum melongena and preparation method of molecular marker

A technology of molecular markers and color genes, which is applied in the fields of molecular biology and eggplant breeding, can solve the problems of no DNA molecular marker reports, no clear genetic mechanism, etc., and achieve the effects of convenient and fast detection, stable results, and improved selection efficiency

Active Publication Date: 2021-01-05
WUHAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there have been some analyzes on the genetic correlation of eggplant sepal color, the genetic mechanism has not been clarified, and there are no reports of related linked DNA molecular markers

Method used

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  • CAPS molecular marker closely linked with sepal color trait gene of solanum melongena and preparation method of molecular marker
  • CAPS molecular marker closely linked with sepal color trait gene of solanum melongena and preparation method of molecular marker
  • CAPS molecular marker closely linked with sepal color trait gene of solanum melongena and preparation method of molecular marker

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Example 1: Construction and DNA Extraction of Eggplant Sepal Color Genetic Population

[0031] (1) Group construction

[0032] In the embodiment of the present invention, eggplant varieties GP05 (green sepal eggplant) and PP05 (purple sepal eggplant) are used as parents, and hybridization (orthogonal) and selfing are carried out in spring and autumn greenhouses to prepare genetic populations, namely P1, P2, and F2. The genetic population was set in the greenhouse at the same time, 30 strains of P1, P2, and Fl were not segregated, and 240 strains of F2 were isolated, and they were cultivated and managed conventionally.

[0033] Eggplants from both parents and genetic populations reached commercial maturity and were identified after harvest. According to the eggplant germplasm resource description specification, two eggplant pairs that reached commercial maturity, conformed to the characteristics of the strain, and developed normally were harvested from each plant. Photo...

Embodiment 2

[0037] Example 2: Searching for green sepal eggplant-associated regions by high-throughput sequencing

[0038] (1) High-throughput sequencing: Sepal color-related SNP screening: Sequencing using the Hiseq2500 sequencing platform, the extracted eggplant parents GP05, PP05, and eggplant green sepal progeny mixed pool BSAg, eggplant purple sepal progeny mixed pool BSAp were high-throughput Quantitative sequencing.

[0039] (2) Sequencing raw sequence processing: the raw image data files obtained by high-throughput sequencing are converted into raw sequencing sequences (raw data) through basecalling analysis. Then, according to certain rules, filter the original sequencing sequence obtained by sequencing. The main criteria are as follows: 1) remove the reads pair with adapter; 2) when the ratio of N in the single-end sequencing read is greater than 10%, it needs to be removed For reads; 3) When the number of low-quality (less than 5) bases in the single-end sequencing read exceed...

Embodiment 3

[0043] Example 3: Design and synthesis of polymorphic primers

[0044] According to the 42 scaffold sequence features obtained by high-throughput sequencing that may be associated with green sepal traits, 34 pairs of Caps primers were designed, and BglII, HindIII, MboI, NcoI, NdeI and XbaI were selected as restriction enzymes for identification. Use these primers to perform PCR amplification on the two parental DNAs. After the PCR amplification products are digested with restriction endonucleases, and then go through agarose gel electrophoresis and staining, the size of the bands is discriminated and polymorphisms are screened. sex primers. The primer sequences were synthesized by Beijing Aoke Biotechnology Co., Ltd., and the restriction enzymes were purchased from NEB Company.

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Abstract

According to the invention, whole genome polymorphic sites of green sepal solanum melongena and purple sepal solanum melongena are analyzed through a high-throughput QTL-seq technology, and SNP sitesare converted into CAPS molecular markers; and one CAPS molecular marker EGC277 capable of effectively discriminating the green sepal solanum melongena is obtained from the CAPS molecular markers, andthe sepal color of the solanum melongena can be predicted through PCR amplification, PCR product enzyme digestion and electrophoresis detection analysis, so that a single plant of the green sepal solanum melongena can be accurately and quickly screened.

Description

technical field [0001] The invention belongs to the field of molecular biology and eggplant breeding, and more specifically relates to a CAPS molecular marker related to color traits of eggplant sepals, and also relates to a preparation method of CAPS molecular markers related to color traits of eggplant sepals, and also relates to a color trait related marker of eggplant sepals Use of CAPS molecular markers. Background technique [0002] Eggplant (Solanum melongena L.) is a herbaceous plant of the family Solanaceae, which is widely cultivated all over the world. It is listed as the fourth largest vegetable crop by the Food and Agriculture Organization of the United Nations. As an important genetic marker, molecular markers can be used in the construction of genetic maps of horticultural crops, QTL (quantitative trait locus) analysis, molecular marker-assisted selection breeding and other research. Among them, molecular markers such as SSR (simple sequence repeats), AFLP (a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12Q1/6811C12N15/11
CPCC12Q1/6811C12Q1/6858C12Q1/6895C12Q2600/13C12Q2600/156C12Q2531/113C12Q2535/122C12Q2521/301C12Q2565/125
Inventor 陈霞张洪源张敏谈杰黄树苹
Owner WUHAN ACADEMY OF AGRI SCI
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