Molecular marker SVhd1 closely linked to growth-period gene of millet
A technology of millet growth period and molecular markers, applied in the field of molecular biology, can solve the problem of gene mapping research that has not been reported in the literature
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Embodiment 1
[0034] Embodiment 1: Construction of millet RIL population
[0035] Male parent: Ningxia Zaogu, precocious (short growth period, 104 days). Female parent: Wudang 2 (WD-2), late maturing (long growth period, 116 days). The male parent and the female parent were crossed to obtain F1, and the F1 generation obtained 215 recombinant inbred lines RIL strains (F6 generation) through the single-grain method. Ningxia Zaogu and WD-2 can be purchased from Shenzhen Huada Agriculture and Circular Economy Technology Co., Ltd.
Embodiment 2
[0036] Example 2: Extraction of Genomic DNA
[0037] 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:
[0038] (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.
[0039]Among them, the formula of 1.5×CTAB is as follows (1L):
[0040] CTAB 15g 1mol / L Tris.Cl (pH 8.0) 75mL 0.5mol / L of EDTA 30mL NaCl 61.4g
[0041] Add deionized water to make up to 1 L, and add mercaptoethanol with a final concentration of 0.2% (2 ml) before use.
[0042]...
Embodiment 3
[0047] Example 3: Gene Mapping and Molecular Marker Development
[0048] (1) Genetic map construction
[0049] 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.
[0050] Use MapMaker 3.0 software (Constructing genetic maps with MAPMAKER / EXP 3.0, S Lincoln, M Daly, 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 .
[0051] (2) Gene mapping
[0052] 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 phenot...
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