Molecular marker sisv0832 closely linked to heading date gene in millet

A technology of molecular markers and heading date, applied in the field of molecular biology, can solve the problem that there is no literature report on the heading date of millet

Inactive Publication Date: 2016-06-01
深圳华大基因农业控股有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, rice is more researched on the heading date, but there are basically no literature reports on the research on the heading date of millet

Method used

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  • Molecular marker sisv0832 closely linked to heading date gene in millet
  • Molecular marker sisv0832 closely linked to heading date gene in millet
  • Molecular marker sisv0832 closely linked to heading date gene in millet

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Construction of millet F2 generation segregation population

[0042] Male parent: Kangnabujing, high plant type, long and narrow flag leaves, red bristles, red glumes, fertile, greenish leaves, yellow-white pollen, late heading stage. The male parent is Zhang Gu No. 1 seed.

[0043] Female parent: Not resistant to Nabujing, short plant type, short and wide flag leaves, green setae, green glumes, partly sterile, yellowish leaf color, brown pollen, early heading stage. The female parent is the seed of millet A2 male sterile line.

[0044] F2 population construction: male parent and female parent are crossed to obtain F1 generation (the heading stage of F1 is late), and F1 is self-crossed to obtain F2. Among them, F1 is the No. 3 seed of Zhang Zagu. A total of 480 individual plants of the F2 generation were obtained.

[0045] See the Chinese patent application "Molecular marker SIsv0372 closely linked to the herbicide resistance gene of millet", publication ...

Embodiment 2

[0046] Example 2: Extraction of parental and F1 generation, F2 generation individual genomic DNA

[0047] Genomic DNA of the parents, the F1 generation, and 480 F2 generation individuals in Example 1 were extracted by the CTAB method, and the specific methods were as follows:

[0048] (1) Weigh 1.0g of fresh leaves, cut them into pieces and put them into a mortar, grind them with liquid nitrogen, add 3mL1.5×CTAB, grind them into a homogenate and transfer them to a 15mL centrifuge tube, then add 1mL1.5×CTAB into the mortar Wash with ×CTAB 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.

[0049] Among them, the formula of 1.5×CTAB is as follows (1L):

[0050]

[0051] Add deionized water to make up to 1 L, and add mercaptoethanol with a final concentration of 0.2% (2 ml) before use.

[0052] (2) After cooling to room temperature, an equal volume of chloroform / isoamyl alcoh...

Embodiment 3

[0057] Example 3: Preparation of Molecular Markers

[0058] Using the genomic DNA of the male parent, the F1 generation, or the F2 generation extracted in Example 2 as a template, PCR amplification was performed with a molecular marker amplification primer pair (SeqIDNo.2 and SeqIDNo.3).

[0059] The PCR reaction system is as follows:

[0060]

[0061]

[0062] The PCR reaction procedure is as follows:

[0063] Pre-denaturation at 94°C for 5 minutes; denaturation at 94°C for 30 seconds, annealing at 60°C for 30 seconds, extension at 72°C for 40 seconds, and 35 cycles; final extension at 72°C for 3 minutes. PCR amplification products can be stored at 4°C.

[0064] Molecular markers are obtained through the above amplification process, and the amplification product is preferably purified after amplification. Sequenced after purification, the result is shown in SeqIDNo.1.

[0065] Those skilled in the art can understand that the molecular marker can also be obtained by ...

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Abstract

The invention belongs to the field of molecular biology, relates to a molecular marker and especially relates to a molecular marker SIsv0832 closely linked with a Setaria italica L. Beauv. heading stage gene. The molecular marker SIsv0832 has a sequence shown in the Seq ID NO.1. The invention also relates to primers for amplification of the molecular marker SIsv0832, a use of the molecular marker SIsv0832 and the primers in Setaria italica L. Beauv. heading stage gene mapping or Setaria italica L. Beauv. genetic breeding, and a Setaria italica L. Beauv. breeding method. The molecular marker SIsv0832 links a genomic DNA sequence and the Setaria italica L. Beauv. heading stage gene, and is conducive to establishment of a Setaria italica L. Beauv. molecular marker assistant breeding system. A genetic close linkage distance between the molecular marker SIsv0832 and the Setaria italica L. Beauv. heading stage gene is 3.5cM. In Setaria italica L. Beauv. breeding practices and resource and variety identification, the simple, fast and high throughput utilization of the molecular marker SIsv0832 and the primers for amplification of the molecular marker SIsv0832 can be realized.

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 a millet heading date gene. The invention also relates to the primer for amplifying the molecular marker, and the use of the molecular marker and the primer in gene positioning of heading stage of millet or genetic breeding of millet. Background technique [0002] my country is the country of origin of millet (Setariaitalica L.Beauv.) and the concentrated planting country of millet in the world. Millet occupies an important position in my country's national economy and social production, and is of great significance to the construction of dry farming ecological agriculture. Therefore, it is particularly important to accelerate the breeding process of millet. Since millet is only a regionally important crop, the current research methods and methods related to millet are relatively backward. How to apply adva...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/63C12N5/10C12Q1/68
Inventor 张耕耘全志武夏秋菊倪雪梅
Owner 深圳华大基因农业控股有限公司
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