Indel Molecular Markers Related to Thousand-grain Weight of Wheat and Its Application
A technology of molecular markers and thousand-grain weight, which is applied in the fields of genetic engineering and molecular biology, can solve the problems affecting the thousand-grain weight of wheat
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Embodiment 1
[0024] Example 1 Development of Wheat TaGSK1 Gene Functional Marker
[0025] According to the sequence of the wheat TaGSK1 gene (Genbank: AF525086) published on NCBI, a pair of Indel primers with polymorphisms between Yumai 8679 and Heshangmai were designed by using PrimerPremier 5.0 software. Screening among varieties was carried out, and its effect on phenotype was further analyzed in recombinant inbred line populations (RILs) and natural populations, and a pair of primers with specific differences were obtained. The primer sequences are as follows:
[0026] Upstream primer F: 5'-CGCAAATCAAAGCTCACC-3'(Seq ID No.1)
[0027] Downstream primer R: 5'-CCATCCTACAAAGGCACA-3' (Seq ID No.2)
Embodiment 2
[0028] Example 2 Correlation between different genotypes of wheat TaGSK1 gene and grain weight
[0029] 1. Determination of thousand-grain weight of wheat grains
[0030] Randomly select 1000 whole wheat grains, repeat twice, weigh with a one-thousandth balance, and record the average value of the two times as the final thousand-grain weight value.
[0031] 2. SDS-Tris saturated phenol method to extract genomic DNA from wheat leaves
[0032] (1) Take 2g of fresh young leaves, grind them into fine powder with liquid nitrogen, and place them in a 2ml sterilized centrifuge tube.
[0033] (2) Add 1.2ml DNA extraction buffer (200mM Tris-Cl, pH8.0, 250mMNaCl, 25mM EDTA, pH8.0, 0.5% SDS, 2% β-ME, mix well, add β-ME freshly before use) .
[0034] (3) 55°C water bath for 45 minutes, with intermittent shaking during the period to fully extract.
[0035] (4) Centrifuge at 12000 rpm for 10 min at room temperature.
[0036] (5) Transfer the supernatant to a new 2ml sterilized centrifu...
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