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Mung bean InDel molecular marker detection primer group and application thereof

A technology of molecular markers and detection primers, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problem of low polymorphic information content, achieve high polymorphism, good repeatability, Rich number of effects

Active Publication Date: 2021-06-15
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Through the second-generation sequencing technology, researchers have developed some new mung bean molecular markers, but the polymorphic information content (PIC) of these markers is relatively low (Tangphatsornruang et al., 2009; Chen et al., 2015)

Method used

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  • Mung bean InDel molecular marker detection primer group and application thereof
  • Mung bean InDel molecular marker detection primer group and application thereof
  • Mung bean InDel molecular marker detection primer group and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1I

[0019] Embodiment 1InDel mark development

[0020] After extracting the genomic DNA of mung bean varieties Sulu 1 and V2709, using illumine SBS constructed the library and performed whole genome sequencing using illumine MiSeq sequencing. After removing adapters and filtering, paired-end sequence data of 20,526Mb and 17,944Mb were obtained for Sulu 1 and V2709, respectively. Use the workflow of mInDel software to develop InDel markers, and the expected InDel size parameters are set to 20-500bp. As a result, 1809 pairs of candidate InDel-labeled primers were generated.

[0021] The resulting InDel-marked primer sequences were compared to the mung bean reference genome sequence (GenBank assembly accession: GCA_000180895.1) using BLAST to obtain the physical positions of these primers. According to the comparison results, filter out some candidate markers according to the following four conditions: 1) There is a mismatch at the 3' end of the primer; 2) The front primer and th...

Embodiment 2

[0065] Embodiment 2 mung bean genetic diversity analysis

[0066] Randomly select 24 polymorphic InDel markers in Table 1, including M0082, M0110, M0115, M0130, M0183, M0189, M0310, M0366, M0550, M0616, M0793, M0826, M0841, M1160, M1229, M1252, M1308, M1309, For M1344, M1351, M1355, M1366, M1489 and M1516, a 0-1 matrix was formed for the genotyping band data of natural populations composed of 40 mung bean varieties (or germplasm resources), and the polymorphic markers among various germplasm materials were counted. number. And use powermarker to calculate the Nei’s genetic distance among various qualitative materials, and construct a phylogenetic tree according to the distance matrix, and perform Neighbor-Joining cluster analysis, and the generated results are as follows figure 2 As shown, 40 mung bean materials can be divided into two groups.

Embodiment 3

[0067] Example 3 Authenticity Identification of Mung Bean Hybrid Progeny

[0068] The mung bean variety Sulu No. 1 was used as the female parent and V2709 was used as the male parent for hybridization. The obtained seeds are planted as a single plant after maturity. Take the leaves of each hybrid plant to be tested, and use the CTAB method to extract DNA. Using the DNA of each hybrid plant to be tested and the DNA of the two parents as templates, PCR amplification was performed with InDel-marked M0679 primers. The obtained PCR products were subjected to electrophoresis in 3% agarose gel. If the hybrid plant to be tested has two bands, one of which is the same size as Sulu 1 and the other is the same size as V2709, then the plant is a true hybrid progeny. If the hybrid plant to be tested only gets one band in the same phase as Sulu No. 1, it is a false hybrid progeny.

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Abstract

The invention discloses a mung bean InDel molecular marker detection primer group and application thereof, and belongs to a plant molecular marker technology. The mung bean InDel molecular marker detection primer group is uniformly distributed on 11 chromosomes, and the primer group sequence is shown as SEQ ID NO.1-1794. The mung bean InDel molecular marker detection primer group is rich in quantity, high in polymorphism, stable in heredity and good in repeatability, and can be used for relatively simple agarose electrophoresis detection, so that the efficiency is greatly improved; and the invention also discloses a kit and a detection method of the InDel molecular marker, and the method is simple and high in success rate, can avoid the problem of fuzzy subsequent analysis caused by specificity and complexity, and has wide application prospects.

Description

technical field [0001] The invention relates to the technical field of plant molecular markers, in particular to a mung bean InDel molecular marker detection primer set and its application. Background technique [0002] Mung bean (Vigna radiate L.) is a diploid plant with 2N=22 chromosomes in its genome. Compared with some model crops, there are not many studies on molecular genetics and genomics related to mung bean, but there has been some development in recent years. As an important tool for genetic analysis and gene mapping, the development of molecular markers for mung bean has also made considerable progress. In recent years, the whole genome sequence of mung bean variety VC1973A has been released (Kang et at., 2014), which is a major advance in mung bean genetics and will be useful for future molecular marker development, gene mapping, and gene function analysis. Bring convenience. Isemura et al. (2012) used the wild mung bean JP211847 and the cultivated mung bean ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2531/113C12Q2565/125C12Q2535/122
Inventor 陈景斌李群三王茜陈新袁星星薛晨晨闫强吴然然林云刘金洋
Owner JIANGSU ACAD OF AGRI SCI
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