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Indel molecular marker related to wheat thousand seed weights and 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

Inactive Publication Date: 2016-02-24
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no research report on the influence of this gene on the thousand-grain weight of wheat

Method used

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  • Indel molecular marker related to wheat thousand seed weights and application
  • Indel molecular marker related to wheat thousand seed weights and application
  • Indel molecular marker related to wheat thousand seed weights and application

Examples

Experimental program
Comparison scheme
Effect test

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 Primer Premier 5.0 software. Screening among wheat 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' (SeqIDNo.1)

[0027] Downstream primer R: 5'-CCATCCTACAAAGGCACA-3' (SeqIDNo.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 of DNA extraction buffer (200mM Tris-Cl, pH 8.0, 250mM NaCl, 25mM EDTA, pH 8.0, 0.5% SDS, 2% β-ME and mix well, β-ME is added fresh just 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 ster...

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Abstract

The invention discloses an Indel molecular marker related to wheat thousand seed weights. On the basis of research of predecessors, the wheat TaGSK1 gene sequence disclosed by NCBI is used as a template design polymer, according to the sequence difference between wheat varieties yamai 8679 and shangmai with the different thousand seed weights, the Indel molecular marker related to the wheat thousand seed weights is developed, the different equipotential variations of the marker can be used for explaining 7.9-11.8% of the thousand seed weight phenotypic variation, strips with the size of amplification products being 624 bp have the synergistic effect on increase of the thousand seed weights, and strips with the size of amplification products being 609 bp have the synergistic effect on reduction of the thousand seed weights. The development of the molecular marker provides a feasible path for high-yield wheat molecule assistant breeding.

Description

technical field [0001] The invention relates to the fields of genetic engineering and molecular biology, in particular to an Indel molecular marker related to wheat thousand-grain weight and its application. Background technique [0002] Wheat is one of the most important food crops in the world. With the increase of population, the reduction of arable land and the continuous increase of food production costs, high-yield and super-high-yield breeding has become an urgent problem to be solved in wheat breeding in my country. Spike number, grain number per panicle and thousand-grain weight are the three elements of wheat yield. Grain weight is a quantitative trait jointly regulated by major genes and minor genes. Therefore, it is important to understand the genes involved in the regulation of grain weight in the breeding of high-yielding wheat varieties. Significance. [0003] Glycogen synthase kinases (GSKs) are highly conserved serine / threonine protein kinases. The GSKs fam...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
CPCC12Q1/6895C12Q2600/124C12Q2600/156
Inventor 朱晓峰常成张海萍司红起卢杰马传喜
Owner ANHUI AGRICULTURAL UNIVERSITY
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