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Combination of MNP Core Primers for Molecular Identification of Eggplant DNA Varieties and Its Application

A technology of molecular identification and primer combination of varieties, applied in the field of molecular biology, can solve problems such as hindering the development of MNP sites

Active Publication Date: 2021-06-11
WUHAN ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the lack of eggplant resequencing data published in NCBI and other databases, the development of MNP sites is hindered

Method used

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  • Combination of MNP Core Primers for Molecular Identification of Eggplant DNA Varieties and Its Application
  • Combination of MNP Core Primers for Molecular Identification of Eggplant DNA Varieties and Its Application
  • Combination of MNP Core Primers for Molecular Identification of Eggplant DNA Varieties and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Screening and Verification of MNP Core Marker Primer Combinations for Molecular Identification of Eggplant DNA Varieties (1) Screening of Eggplant MNP Primer Marker Combinations

[0025]The publicly released SME_r2.5.1 sequence was used as the eggplant reference genome (http: / / eggplant.kazusa.or.jp / ), with a total length of 850,044,630bp, assembled to the contig level; two varieties (PnP05) provided by the Wuhan Academy of Agricultural Sciences were used. and GnP05, cultivar name link https: / / doi.org / 10.1371 / journal.pone.0179491) genome resequencing data, 8 NCBI eggplant resequencing data (NCBI accession numbers SRR5796636 to SRR5796643), and publicly available Eggplant resequencing data NCBI accession numbers are GB Accession No. 30075032 to 3007503258; No. 30075103 to 30075116. According to the above sequencing data, 522 MNP marker sites were screened for comparative analysis, and primers were designed (see Table 1 for details of primers).

[0026] (2) Correctness an...

Embodiment 2

[0032] Construction of eggplant standard DNA fingerprints by combining MNP core marker primers for molecular identification of eggplant DNA varieties

[0033] 1. DNA extraction and quality assessment

[0034] The genomic DNA of all eggplant varieties used to construct eggplant fingerprint database was extracted by conventional CTAB method, and the quality of DNA was detected by agarose gel and UV spectrophotometer after RNase treatment. If the DNA band is single and has no obvious degradation, A260 / 280 detected by UV spectrophotometer is between 1.8-2.0, and A260 / 230 is between 1.8-2.0, there is no obvious light absorption at 270nm, and the DNA concentration is greater than 50ng / ul, it means that The genomic DNA meets the relevant quality requirements and can be used for subsequent experiments.

[0035] 2. MNP marker detection

[0036] Using the synthesized 522 eggplant MNP marker primers, 80 eggplant inbred strains provided by the Vegetable Research Institute of Wuhan Acade...

Embodiment 3

[0040] MNP core marker primer combination for molecular identification of eggplant DNA varieties to detect the authenticity of eggplant varieties

[0041] 1. Determination of the discrimination of eggplant varieties marked by MNP primers and the determination of the authenticity test results of eggplant varieties

[0042] The 80 eggplant inbred line samples in Example 2 were compared in pairs, and the differences in the MNP marker sites were analyzed, and 3160 pairs of comparison results were obtained. The difference in distance was between 0% and 83.69%, with an average difference of 44.16%, of which 8 pairs had 0 differences, accounting for 0.25% of the total sample pairs. Using the developed marker detection method, 99.75% of eggplant varieties could be distinguished ( figure 2 ), indicating that the developed eggplant MNP marker has high polymorphism and strong cultivar discrimination. Therefore, 522 loci can be fully applied to the authenticity identification of eggpla...

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Abstract

The invention belongs to the technical field of molecular biology, and specifically discloses a combination of MNP core primers for molecular identification of eggplant DNA varieties and applications thereof. The primer combinations are shown in Table 1. The invention provides a primer combination composed of 522 MNP core primers, which is compatible with the second-generation sequencing platform, has the characteristics of high efficiency, accuracy, and high reproducibility, and meets the requirements for constructing standard eggplant fingerprint data; , can accurately identify the authenticity of the eggplant sample to be tested; the analysis of eggplant high-generation inbred line materials shows that this MNP site has high polymorphism and repeatability, can distinguish different eggplant groups, and has a wide range of applications .

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and particularly relates to an MNP core primer combination for molecular identification of eggplant DNA varieties and its application. [0002] technical background [0003] Eggplant (Solanum melongena L.) is a vegetable crop of the Solanaceae family, which is widely cultivated all over the world. It is the fourth largest vegetable crop in the world and one of the main vegetable crops in my country. According to the United Nations Food and Agriculture Organization (FAO) statistics in 2018, the global eggplant production is about 50 million tons, of which Asian eggplant accounts for more than 90% of the total production. The annual planting area in China is about 800,000 hectares, and the total production accounts for the global total. more than 50% of the output. my country is the secondary origin of eggplant, with a long history of cultivation and abundant germplasm resources. The nati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6806C40B50/06C12N15/11G16B20/40G16B25/20G16B30/10G16B40/10G16B40/30G16B45/00G16B50/00
CPCC12Q1/6806C12Q1/6895C12Q2600/156C40B50/06G16B20/40G16B25/20G16B30/10G16B40/10G16B40/30G16B45/00G16B50/00C12Q2525/191C12Q2531/113
Inventor 张洪源张敏谈杰黄树苹陈霞
Owner WUHAN ACADEMY OF AGRI SCI
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