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Molecular marker SVmc3 closely linked with husked millet color genes

A technology of molecular markers and color genes, which is applied in the field of molecular biology, can solve the problems that there are no literature reports on the mapping of beige-color related genes of millet

Inactive Publication Date: 2017-06-09
SHENZHEN BGI AGRI & CYCLE ECONOMIC TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no literature reporting the mapping of beige-related genes in millet

Method used

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  • Molecular marker SVmc3 closely linked with husked millet color genes
  • Molecular marker SVmc3 closely linked with husked millet color genes
  • Molecular marker SVmc3 closely linked with husked millet color genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: Construction of millet RIL population

[0028] Male parent: JZG, Huangmi. Female parent: MJG, black rice. The male parent and the female parent were crossed to obtain F1, and the F1 generation obtained 281 recombinant inbred lines RIL strains (F6 generation) through the single-grain method.

[0029] Millet seeds of JZG, MJG, F1 and recombinant inbred lines can be purchased from Shenzhen Huada Agriculture and Circular Economy Technology Co., Ltd. and the Millet Research Institute of Zhangjiakou Academy of Agricultural Sciences.

Embodiment 2

[0030] Example 2: Extraction of Genomic DNA

[0031] For the millet RIL population obtained in Example 1, use the CTAB method to extract the genomic DNA of the parents and the individual plants of the RIL population respectively. The specific methods are as follows:

[0032] (1) Weigh 1.0g of fresh leaves, cut them into pieces and put them in a mortar, grind them with liquid nitrogen, add 3mL 1.5×CTAB, grind them into a homogenate and transfer them to a 15mL centrifuge tube, then add 1mL 1.5×CTAB into the mortar Rinse and transfer to a centrifuge tube. After mixing, place in a water bath at 65°C for 30 minutes, and shake slowly from time to time during this period.

[0033] Among them, the formula of 1.5×CTAB is as follows (1L):

[0034] CTAB 15g 1mol / L Tris.Cl (pH 8.0) 75mL

[0035] 0.5mol / L of EDTA 30mL NaCl 61.4g

[0036] Add deionized water to make up to 1 L, and add mercaptoethanol with a final concentration of 0.2% (2 ml) b...

Embodiment 3

[0042] Example 3: Gene Mapping and Molecular Marker Development

[0043] (1) Genetic map construction

[0044] For the genomic DNA of the RIL individual obtained in Example 2, RAD-seq-based genotyping technology (http: / / www.bioon.com.cn / server / show_product.asp?id=12291) was used for individual RIL population Genotyping was performed to obtain genotype data for the RIL population.

[0045] Use MapMaker 3.0 software (Constructing genetic maps with MAPMAKER / EXP 3.0, S Lincoln, MDaly, E Lander-Cambridge, MA: Whitehead Institute, 1992, which is incorporated herein by reference) to draw a genetic linkage map to obtain a genetic linkage map .

[0046] (2) Gene mapping

[0047]For the RIL population obtained in Example 1, the individual phenotypes of the RIL population, which are similar to the paternal traits, are marked as a, those similar to the maternal traits are marked as b, and the traits are between the male parent and the female parent Denote it as h. Obtain the phenotyp...

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Abstract

The invention belongs to the field of molecular biology, and relates to a molecular marker, in particular to a molecular marker closely linked with husked millet color genes. The molecular marker contains a sequence as shown in SEQ ID NO.1. The invention also relates to a primer pair for extending the molecular marker, a detection method of the molecular marker, an application of the molecular marker or the primer pair in millet assisted breeding or location or detection of the husked millet color genes, a millet assisted breeding method, and a method for screening the molecular marker. The molecular marker provided by the invention combines genome DNA sequence and the husked millet color gene, so that the establishment of the millet molecular marker assisted breeding system is facilitated; and a genetic close linkage distance of the molecular marker and the husked millet color gene is 1.2cM. The molecular marker and the molecular marker extended primer can be conveniently and rapidly used for the millet breeding practice and the resource and variety identification in a high flux manner.

Description

technical field [0001] The invention belongs to the field of molecular biology and relates to a molecular marker, in particular to a molecular marker closely linked with the color gene of millet grains. The present invention also relates to the primer of the molecular marker, the use of the molecular marker in the mapping of the color gene of millet or millet genetic breeding, a method for mapping the color gene of millet, and a method for breeding of millet. Background technique [0002] Millet (Setaria italica (L.) Beauv.) originated in China and is a traditional dominant crop, staple food crop and drought-resistant crop. Millet is resistant to drought and barrenness, has high water use efficiency and wide adaptability. It not only plays an important role in the current dry farming ecological agriculture, but also is an important strategic reserve crop for the increasingly serious water shortage. The millet after millet hulling is rich in nutrients and balanced in various...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13
Inventor 倪雪梅蔡雪梅雷雪静雍建朋肜君
Owner SHENZHEN BGI AGRI & CYCLE ECONOMIC TECH
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