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Molecular marker closely linked with millet tillering character, primer and application

A technology of molecular markers and millet, applied in the field of molecular biology, can solve the problems of undiscovered molecular markers, etc., and achieve the effect of accelerating the process of genetic selection and improvement

Inactive Publication Date: 2017-12-15
山西省农业科学院谷子研究所 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, no molecular markers closely linked to millet tillering traits have been found in the prior art.

Method used

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  • Molecular marker closely linked with millet tillering character, primer and application
  • Molecular marker closely linked with millet tillering character, primer and application
  • Molecular marker closely linked with millet tillering character, primer and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Obtaining method of molecular marker MRI381

[0035] The female parent of the tillering trait Ai Ninghuang was used as the female parent; the non-tillering trait Jingu 21 was used as the male parent, and the F 1 , single plant F 1 543 F2 segregating populations were obtained by selfing. Using hole sowing method, the F 2 Planted in the experimental field of Millet Research Institute of Shanxi Academy of Agricultural Sciences, the row length is 6m, the row spacing is 0.33m, the plant spacing is 15cm, and each parent has one row. The cultivation management measures in the field test are consistent with the local field management. Seedling pair of parents and 543 F 2 Individual plants were sampled. F 2 The tillering survey was carried out on a single plant, and Microsoft Excel 2010 was used for statistical analysis of the data.

[0036] Using the RAD-seq method, the parental and 543 F 2 The genomic DNA of a single plant is tested for DNA quality, the quali...

Embodiment 2

[0048] Embodiment 2: millet F 2 Population library sequencing

[0049] The millet F obtained in embodiment 1 2 The group database construction method is as follows:

[0050] (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.

[0051] The 1.5×CTAB formula is as follows (1L): CTAB 15g, 1mol / L Tris.Cl (pH 8.0) 75mL, 0.5mol / L EDTA 30mL, NaCl 61.4g, add deionized water to 1L, before use Mercaptoethanol was added to a final concentration of 0.2% (2 ml).

[0052] (2) After cooling to room temperature, add an equal volume of chloroform / isoamyl alcohol (24:1, v / v) and mix gently until the lower layer turns dark green.

[0053] (3) Centrifu...

Embodiment 3

[0074] Example 3: Extraction of Genomic DNA

[0075] For the millet F obtained in Example 1 2 Group, use CTAB method to extract parents and F 2 Genomic DNA of individual plants in the population, the specific method is as follows:

[0076] (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.

[0077] The 1.5×CTAB formula is as follows (1L): CTAB 15g, 1mol / L Tris.Cl (pH 8.0) 75mL, 0.5mol / L EDTA 30mL, NaCl 61.4g, add deionized water to 1L, before use Mercaptoethanol was added to a final concentration of 0.2% (2 ml).

[0078] (2) After cooling to room temperature, add an equal volume of chloroform / isoamyl alcohol (24:1, v / v) and mix gen...

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Abstract

The invention belongs to the technical field of molecular biology, and particularly discloses a molecular marker closely linked with millet tillering character. The molecular character MRI381 is located in the range of 31021885bp to 31095359bp of the seventh chromosome of millet. The invention further discloses an amplification primer and application of the molecular marker. The molecular marker can be used for excellent typing on millet tillering, can be used for molecular marker assistant breeding of millet tillering character, and has important theoretical and practical guiding significance on accelerating genetic breeding and improving process of millet varieties.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a molecular marker closely linked with millet tillering traits, a primer and an application thereof. Background technique [0002] my country is the origin of millet and the country with the largest cultivated area and the highest yield in the world, accounting for about 80% of the world's total. At the same time, my country is also the country with the largest number and diversity of millet genetic resources. Tillering is a special branch of monocotyledonous plants, and it is also an agronomic characteristic that plays an important role in the growth and development of millet plants. The utilization of tillering germplasm resources of millet is beneficial to reduce plant height and improve lodging resistance of millet, and it can also be achieved by relying on large groups. High-yielding, relying on tillering to form ears to achieve precision sowing withou...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13
Inventor 王军杜晓芬杜国华王智兰邹洪锋杨慧卿郭二虎彭建祥韩芳袁峰张林义彭书忠
Owner 山西省农业科学院谷子研究所
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